Docstoc

SEROLOGY

Document Sample
SEROLOGY Powered By Docstoc
					                                              Policy # MI/SER/v25              Page 1 of 2
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                      Subject Title: Table of Contents
Issued by: LABORATORY MANAGER                 Original Date: March 14, 2001
Approved by: Laboratory Director              Revision Date: January 31, 2011
                                              Annual Review Date: January 10, 2011

                                                         SEROLOGY MANUAL
                                                         TABLE OF CONTENTS

Immucor Capture-CMV ........................................................................................................................... 3
Architect System for HBsAg, HBsAb, HBeAg, HBeAb, HBcAb-IgM, HBcAbTotal, HAV-IgM,
HCVAb, CMV IgG, HIV-1/2, Rubella IgG,Syphilis TP Ab,HBsAg Qualitative and HBsAg Qualitative
Confirmatory ............................................................................................................................................ 6
AxSYM System for HBsAg, HBsAb, HBcAbTotal, HCVAb, CMV IgG, and HIV-1/2 ..................... 18
AxSYM Heptitis B Surface Antigen Confirmatory Assay .................................................................... 33
Labour and Delivery Hepatitis Virus Serology........................................................................................ 37
Labour and Delivery Human Immunodeficiency Virus (HIV) 1 & 2 Serology ................................... 39
HBsAg , HIV 1/2,HBcAb, HCV Ab & HTLV I/II (cadaver/donor) Serology ..................................... 40
Epstein Barr Virus Viral Capsid Antigen IgG ....................................................................................... 59
Aspergillus Galactomannan Antigen Detection Assay .......................................................................... 68
Human T-Lymphotropic Virus Type 1 (HTLV-1) EIA for TGLN Call Back Only ............................. 81
Infectious Mononucleosis Heterophile Antibodies .................................................................................. 90
Syphilis Screening ................................................................................................................................. 92
Varicella-Zoster Virus IgG .................................................................................................................... 95
West Nile Virus IgM EIA Test ............................................................................................................ 101
APPENDIX I - SEROLOGY TEST SCHEDULE ................................................................................ 107
APPENDIX II - List of Tests Referred Out to Other Laboratories ....................................................... 108
APPENDIX III - SHIPMENT OF SAMPLES to ST. JOSEPH'S HOSPITAL ...................................... 110
APPENDIX IV - ENTERING SEROLOGY REFER-OUT RESULTS ................................................ 111
APPENDIX V - AUTOVERIFICATION PROCESS ........................................................................... 114
APPENDIX VI - SHIPMENT OF SAMPLES TO HSC ....................................................................... 116
APPENDIX VII - POSTING OF AxSYM RESULTS .......................................................................... 117
APPENDIX VIII - LOOKING UP PREVIOUS HEPATITIS RESULTS IN EPR............................. 118
APPENDIX IX - PRINTING OF PENDING LIST .............................................................................. 119

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                         Page 1
                                             Policy # MI/SER/v25              Page 2 of 2
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                     Subject Title: Table of Contents

APPENDIX X - ENTERING REFERRED TEST RESULTS .............................................................. 120
Record of Edited Revisions ................................................................................................................. 121




TORONTO GIFT OF LIFE NETWORK PROCEDURES:
          TGLN Ordering in Mysis Procdeure
          TGLN Recipient STAT Testing Procedure
          TGLN Call Back Procedure

Labour Delivery Stat HIV HBsAg Instructions




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                    Page 2
                                                                          Policy # MI/SER/02/ v02                     Page 1 of 3
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Immucor Capture-CMV
Issued by: LABORATORY MANAGER                                             Original Date: July 14, 2008
Approved by: Laboratory Director                                          Revision Date:
                                                                          Annual Review Date: January 10, 2011

                                                             Immucor Capture-CMV

     I. Introduction

          The Immucor Capture-CMV is an in vitro qualitative solid phase red cell adherence test system
          for the detection of antibodies (IgG plus IgM) to Cytomegalovirus (CMV) in human serum or
          plasma. Capture-CMV is intended to be used in screening of donors or patients for serological
          evidence of previous infection by CMV.

     II. Specimen Collection

          Serum of plasma can be separated from red cells and stored at 1-10oC for up to 1 week,
          or frozen at -20oC freezer. Sample should not be repeatedly frozen and thawed.
          Gel separation tube for serum collection is not recommended.

     III. Procedure

               a. Reagent preparation:
                      i. Wash solution: 20 mL pHix to 2000 mL of Isotonic Saline. Check the pH using
                         litmus paper with range of 6.0-8.0-pH should be 6.5-7.2. Good for 30 days at
                         store at room temperature.
                     ii. Capture-CMV Microtitration wells: Rigid U-bottom microtitration well coated
                         with glycine-extracted and purified CMV antigen obtained from CMV 169
                         grown in human foreskin fibroblast cells. The wells are enclosed in a foil pouch
                         to which a dessicant and moisture indicator has been added. Store the pouch at
                         1-30oC. Remove the number of strips needed and carefully re-seal pouch to
                         prevent the uptake of moisture. Once pouch is opened, the microtitration wells
                         should be used within 2 weeks if the moisture indicator stays blue. The
                         Microtitration wells should not be used if the moisture indicator turns pink.
                         Microtitration wells removed from the pouch should be used within one (1)
                         hour.
                    iii. Capture-CMV Positive Control Serum(weak), store at 1-10oC.
                    iv. Capture-CMV Negative Control serum, store at 1-10oC.
                     v. Capture LISS: a low ionic strength solution containing Glycine-bromcresol
                         purple dye and sodium azide. Store at 1-10oC.
                    vi. Capture-CMV indicator Red-Cells, store at 1-10oC.
               b. Assay Method:
                     1. Bring reagents to room temperature.
                     2. Remove Capture-CMV Microtitration wells from the pouch.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                   Page 3
                                                                          Policy # MI/SER/02/ v02                     Page 2 of 3
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Immucor Capture-CMV

                          3. Add 2 drops (100 uL ± 10 uL) of Capture-LISS to all test wells. The color will
                              change from purple to blue once the control/specimen has been added.
                          4. Add 1 drop (50 uL ± 5 uL) of Capture-CMV Positive Control Serum (weak) to a
                              designated well.
                          5. Add 1 drop (50 uL ± 5 uL) of Capture-CMV Negative Control Serum to a
                              designated well.
                          6. Add 1 drop (50 uL ± 5 uL) of the first specimen to the third well, continue
                              adding to the designated wells for the rest of the specimens.
                          7. Incubate at 18-30oC for a minimum of 5 minutes, but no more than
                              30 minutes.
                          8. Do daily maintenance on CSW 100 Capture Strip Well Washer:
                               i. Prime to remove bubbles in tubing.
                              ii. Check dispense and aspiration needle.
                             iii. Inspect tubing and manifold needles.
                             iv. Residual saline verification-wash a whole plate(12 strips) by pressing on
                                   ‗Wash‘, after washing is finished visually check what is left on each well by
                                   tilting the plate at an angle.
                              v. Fill PBS and Distilled water reservoirs.
                                   Check off on CSW 100 Maintenance sheet. Empty waste container after
                                   testing is finished.
                          9. Wash plate using program 2 using CSW 100 Capture Strip Well Washer.
                              Automatic washer must be adjusted such that approximately 4-6 uL of saline
                              remains in each well after aspiration. Wells should not be aspirated until they
                              are dry
                          10. Re-suspend Capture-CMV indicator Red-Cells by gently inverting the vial,
                              immediately add 1 drop (50 uL ± 5 uL) of Capture-CMV indicator Red-Cells to
                              each well.
                          11. Load strips into centrifuge. Centrifuge the microtitration wells using program 1
                              (450-600 x g) for 2 minute. Then centrifuge again using program 2 (1000-1400
                              x g) for 1 minute. Remove strip(s) from centrifuge immediately, and leave on a
                              flat surface.
                          12. Place the microtitration wells on an illuminated surface and for adherence or the
                              absence of indicator cells.

     IV. Interpretation of Result

          Negative test:: A button of the Capture-CMV indicator Red-Cells at the bottom of the test well
          with no area of adherence indicates the test sample has no detection CMV antibody and the
          person has not yet been infected with CMV and is presumed to be susceptible to primary
          infection.
          Positive test: Adherence of Capture-CMV indicator Red-Cells to part or all of the reaction
          surface indicates a person with previous current infection and who is presumed to be at risk of
          transmitting CMV infection but who is not necessarily currently contagious.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                   Page 4
                                                                          Policy # MI/SER/02/v02                      Page 3 of 3
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Immucor Capture-CMV

     V. Assay Validation

          Test is valid when:
          Weak Positive Control shows adherence of Indicator Cells over part or all of reaction surface.
          Negative Control shows a button of Red Indicator cells at the bottom of the well with no area
          of adherence


     VI. Quality Control

          One Positive Control and one Negative Control must be included with each run.
          Run Virotrol I as Positive external control and Viroclear as Negative External Control.
          CAP provides external control testing.


  VII. Reference

          Package insert from Capture-CMV kit by Immuno Gamma, version 12/05.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                   Page 5
                                                                          Policy # MI/SER/16/v02                      Page 1 of 11
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System
Issued by: LABORATORY MANAGER                                             Original Date: May 17, 2010
Approved by: Laboratory Director                                          Revision Date: January 31, 2011
                                                                          Annual Review Date: January 10, 2011

     Architect System for HBsAg, HBsAb, HBeAg, HBeAb, HBcAb-IgM, HBcAbTotal, HAV-IgM,
      HCVAb, CMV IgG, HIV-1/2, Rubella IgG,Syphilis TP Ab,HBsAg Qualitative and HBsAg
                                    Qualitative Confirmatory

I.        Introduction

         The Abbott Architect System is a fully automated random access analyser that utilizes a
         chemiluminescent microparticle immunoassay (CMIA) technology with flexible assay protocols,
         refer to as Chemflex®. At first, sample containing either Antigen or Antibody is combined with
         Antigen or Antibody coated paramagnet particles. Antigen or Antibody present in the sample
         binds to the Antibody or Antigen coated paramagnet particles. After washing, acridinium-labeled
         Antigen conjugate or Antibody conjugate is added in the second step. Following another wash
         cycler, Pre-trigger and Trigger Solutions are added to the reaction mixture. The resulting
         chemiluminescent reaction is measured as relative light unit(RLU).A direct relationship exists
         between the amount of Antigen or Antibody in the sample and the RLUs detected by the
         Architect System optics.

II.       Specimen Collection and Processing

          Blood is collected (5 mL for adult and 1 mL for neonates) in a serum separator tube, Sodium
          heparin tube, Sodium citrate tube, ACD tube, CPDA-1 tube or EDTA tube. Sample is centrifuged
          (3,000 g x 10 minutes) and the serum is stored refrigerated until testing. After testing is
          completed, specimens are stored at -700C for 10 years for bone marrow and bone bank (live)
          patients and 3 months at 40C for other patients.
          Re-centrifugation is required for samples that are cloudy, previously frozen, for repeat testing or
          reconstituted (e.g. Proficiency testing samples).
          Note: Cadaveric Donor samples must be tested using approved assays (syphilis by RPR;
          HBsAg & HIV Ab by BioRad EIA; CMV Ab by Immucor)

III.      Reagents and bulk solutions

          Reagents, calibrator, control packs, Pre-Trigger Solution and Probe Conditioning Solution
          for the assays are stored refrigerated. Wash buffer and Trigger Solution are stored at room
          temperature. Once any solution is opened, it is only good for 28 days.

          Assays run in Architect are as follows:
          HBsAg, Ausab, HBcII Ab, HBcIgM, HCV Ab, HIV Ag/Ab, HBeAg, HBeAb, HAV IgG,
          HAV IgM, CMV IgG , Rubella IgG ,Syphilis TP Ab , HBsAg Qualitaitve, and HBsAg
          Qualitative confirmatory.

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                   Page 6
                                                                          Policy # MI/SER/16/v02                      Page 2 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

          Each assay will have a reagent kit, calibration kit and controls kit. All assays are either not
          validated/or not to be used for cadaveric specimens.

          A yellow ‗Caution ‗symbol will appear when Bulk solutions ( Wash Buffer, Trigger
          Solution, Pre-trigger solution), Reaction cells and reagent kit level reach < 20%:

           a. Wash buffer: Pour a new bottle of 975 ml wash buffer into the 10 L Deionised H2O.
              Open bottom door, and connect tubing to the port for the buffer container in Architect. Touch
              ‗RV/Liquid Waste‘ screen. Touch ‗Update supply‘ and follow instruction.
           b. Trigger/Pre-trigger solution: Can still run tests for 1 more day. But if it is a Friday, change
               solution at the end of the shift .Slide whole shelf out, move old bottle to the middle slot. Put
               new bottle in the empty slot, and remove the top on the new bottle. Unscrew the lid and move
               the lid and the tubing of the old bottle into the new bottle and tighten the lid. Make sure that
               the white tubing is pointing straight towards you.
           c. Probe Conditioning Solution: keep at 40C, use for daily maintenance.
           b. Reaction cells: ‗update supply, if 1 bag is added, choose ‗500‘, and ‗Done‘.


IV.       Procedure

      At snapshot screen:
      1. Leave System on ‗Running ‗Mode.
      2. Touch SCC (system control center) and RSH( Robotic sample handler) and ‗Pause‘. The status
          will change from ‗Running‘ to ‗Ready‘. Only when Architect is in ‗Ready‘ mode that the
          Reagent Carousel lid can be opened, and the carousel can be turned.
      3. Check Reagent Load List:
              a. Touch Reagent Icon on Snapshot.
              b. Touch ‗View All‘.
              c. Touch ‗Stability‘: will show available hours left .
              d. Touch‘ Remaining Test ‗: will show how many tests left on each kit. Make sure that
                  there are enough reagents for the whole day.
              e. Touch ‗Assay‘: will show # of same reagents loaded.
      4. Do daily maintenance:
          1) Touch ‗System‘ and ‘Maintenance‘.
          2) 6041 Daily maintenance will show on screen. Highlight this line, touch ‘Perform‘. It will take
              approximately 21 minutes.
          3) ‗Are you sure you want to execute the procedure 6041 Daily maintenance‘, touch ‘OK‘.
          4) Touch ‘Proceed‘ when ‗instruction screen‘ appears.
          5) Then:
                  a. Open Reagent Carousel door.
                  b. Fill cleaning bottle with 25-30 ml of 0.5% Javex (good for 1 month), put in position #
                      1 in the inner ring.
                  c. Close the carousel door.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                   Page 7
                                                            Policy # MI/SER/16/v02            Page 3 of 11
                           Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                    Subject Title: Architect System
        6) Touch ‘Proceed‘.
        7) When finished:
                 a. Open the carousel door
                 b. Remove the cleaning bottle and Probe Conditioning solution bottle from the carousel.
                 c. Cap the Probe Conditioning solution bottle, and store both at 40C.
                 d. If is necessary to replace or removed reagent bottle.
                 e. Close Reagent Carousel door.
        8) Touch ‘Proceed‘ and ‗Done‘.
    5. Load controls-all controls are bar-coded. Check level of controls in each sample cup. If level
        reaches the first mark, it is still good for one more run. Change each sample cup when volume
        is too low, fill up to about 1/3 full. Do not overfill. Label the top with name of each control,
        and Lot#, expiration date and date in use on the side. When control fail-CNTL, repeat with a
        new aliquot. If it still fails, recalibrate and re-run the set of controls.1-2 sd is not a control
        failure. It is a warning that this control is outside 1 sd(Standard Deviation), but still within 2-
        sd-do not need to repeat. But if is 1-3sd, must repeat only that control that fails.
    6. . If only 1 control fails out of the whole set, only repeat that specific one by putting in a new
        carrier.
    7. If new controls are used, must update in Architect the same time. Follow ― to enter new control
        #‘ procedure.
    8. To order ‗Controls‘ for a specific lot #:
             a. ‗Order‘,‘ Control Order‘.
             b. ‘Single Analyte‘
             c. C (carrier) field, scan in Carrier #.
             d. P (position) field, type in position #.
             e. ‘ test‘.
             f. Go to 'Details',type in new lot #.
             g. ‗Add order‘.
    9. Remove top from blood tube, load blood tubes into segment or box. Check level of serum
        against the mark on the segment. If serum is below the level, transfer the serum to sample cup.
        The sample cup can be placed inside blood tube, and load into carrier. The carrier will be
        picked up and scanned and Architect will do all the tests ordered.
    10. Once loaded, the Orange light will come on. Do not touch the segment.
    11. The transfer arm will move the segment to the bar code reader. Once bar codes are read, the
        arm will move the segment back to the original position first. Then will come again to
        pick up the segment and move to the processing centre. Do not put other segment in this
        position, because once sampling is finished, the arm will return this segment to the same
        position again.
    12. Once testing is finished, results will be posted in ‗Result Status‘. Also the ‗Result‘ will be
        flashing as soon as result is posted on the screen.
    13. Touch ‗Result Status‘, ‗SID‘, ‗Select all‘ and ‗Print‘, select ‗Result List Report‘.
    14. ‗Select all‘ and ‗Release‘.



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                   Page 8
                                                                          Policy # MI/SER/16/v02                      Page 4 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

     15. If need to create an order:
                a. Select ‗Order‘,‘ Patient Order‘.
                b. C (carrier) field, scan in Carrier #.
                c. P (position) field, type in position #.
                d. ‘ test‘.
                e. ‗sample details‘.
                f. Type in ‗last name‘.
                g. ‗Done‘
                h. ‗Add Order‘

     16. To order Calibration:
            a. Select ‗Order‘,‘ Calibration‘.
            b. C (carrier) field, scan in Carrier #.
            c. P (position) field, type in position #.
            d. Choose ‘Assay‘.
            e. To check if this is a new calibration lot:
                         i. Go to ‗Assay option‘.
                         ii. If it is a new lot-highlight lot #,
                         iii. Enter new Calibrator lot #.
                         iv. Highlight exp date, enter new exp date.
                         v.Done.
            f. ‗Add Order‘.

     17. To enter NEW Control Lot#:
                a.        Log on as ‗ADMIN‘, pass word‘ ADM‘.
                b.        Architect on ‗Ready‘ mode.
                c.        System
                d.        Configuration
                e.        ‗QC Cal Setting‘- ‘QC single analyte‘
                f.        Choose ‗Assay‘ e.g. HBsAg.
                g.        ‗Configure‘.
                h.        A new screen will come up showing lot #, touch ‗box with bars‘: highlight.
                i.        ‗New lot-copy data‘ from the drop down menu.
                j.        Highlight lot #, enter new lot #.
                k.        Highlight exp date, enter new exp date.
                l.        ‗Default‘.
                m.        ‗Done‘.
     18. Once results are posted, the ‗Result‘ field will be flashing. Touch‘ Result‘,
         ‘Result review‘, ‘Print‘, Use ↓Arrow to ‗Results List Report‘, and ‗Done‘.
     19. Underline + HbsAg,+ HbcAb IgG,+ HbcAb IgM, borderline HbeAg and HbeAb,
         borderline and negative Rubella. Follow reporting chart.


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                   Page 9
                                                                          Policy # MI/SER/16/v02                      Page 5 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

      STAT testings:

      For needle stick incident:
      a. Do HBsAg, HCVAb and HIV ½ Ag/Ab for source patient.
      b. Do HBsAb and HCV Ab for staff.
      c. Cadaveric specimen from outside clients(e.g. Forensic Pathology lab)- specimen is tested in
          Architect first,which gives a faster turn around time. Specimen then repeated again using
          BioRad HbsAg and HIV kits the next working day.

      For Case room patients that have no previous prenatal testing done: Do HBsAg and HIV1/2
      Ag/Ab when requested. Also order ‗8Ref‖ and send out ~1.5 ml of the serum to PHL for HIV
      testing. Enter PHL HIV Ab result under ‗8 Ref‘.

     HBsAg Qualitative Confirmatory:

     1. Do confirmation when HBsAg quantitative assay value is <2.00 IU/ml,or if >2.00 and HBc Ab
         is negative. If HbsAg Quantitative assay is >250 IU/ml,make a 1:500 dilution of specimen
         first( 2 ul specimen + 1000 ul Sample Diluent).
     2. ‗Order‘, ‘Patient Order‘.
     3. Scan in C:# , type in P: #, and scan in SID #.
     4. Touch ‗HBsAgQ%‘ under ‗Panel‘- HBsAg% Neut, HBsAg Qcf1 and HBsAg Qcf2 will be
         highlighted.
     5. Touch ‗Sample Details‘ and type in patient‘s last name, and ‗Done‘.
     6. Touch ‗Add order‘.
     7. ‗Order‘, ‘Control Order‘.
     8. Scan in C:# , type in P: #, and scan in SID: Positive Control. Use Positive Control from HBsAg
         Qualitative Control kit.
     9. Touch ‗HBsAgQ%‘ under ‗Panel‘- HBsAg% Neut, HBsAg Qcf1 and HBsAg Qcf2 will be
         highlighted.
     10. Touch ‗Add order‘.
     11. Load specimen and positive control.
     12. HBsAg% Neut result will be calculated when done. Print report.
     13. If HBsAg% Neut is >50%, it is a confirmed positive for HBsAg.
     14. If HBsAg% Neut is <50%:
              a. . HbsAgQ(e.g. >500)→need dilution(1:500-2 ul specimen + 1000 ul Sample
                  Diluent),and repeat confirmation.
              b. HBsAgQ value is low(e.g. <10)→did not confirmed-negative for HBsAg.
      Print‘ Sample report‘.

     Back up files:
     1. Done weekly: back up to Architect f: drive-‗System‘, ‘Utilities‘, ‗F4-create back up‘ and
        ‗Done‘.
     2. Done monthly :back up to CD-‗System‘, ‘Diagnosis‘ , module 5(computer),
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 10
                                                                          Policy # MI/SER/16/v02                      Page 5 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

          ↓ to ‗ ‗Utilities‘,‘6004 copy back up software‘, ‘perform‘, ‘OK‘, ‘Proceed‘ ,‘type in
          1-copy back up software from f: drive to CD‘, ‘continue‘ ‘put in CD‘, ‘Proceed‘, will ask to
          remove CD once finished copying. ‘Done‘.

V.        Cut-Off values:

           Assay                  Neg                       Pos          Gray Zone
           HBsAg                  <0.05 IU/ml               ≥ 0.05 IU/ml
           HBsAb                  <10.00 mIU/ml             ≥ 10.00 IU/L
           HBcAbII                <1.00 S/CO                ≥ 1.00 S/CO
           HBcII IgM              <1.00 S/CO                ≥ 1.00 S/CO
           HBeAg                  <1.00 S/CO                ≥1.00 S/CO
           HBbeAb                 >1.00 S/CO                ≤1.00 S/CO
           HCV Ab                 <1.00 S/CO                ≥1.00 S/CO
           HIV 1&2                <1.00 S/CO                ≥1.00 S/CO
           HAV IgM                <0.08 S/CO                >1.20 S/CO 0.08-1.20 S/CO
           HAV IgG                <1.00 S/CO                ≥1.00 S/CO
           Rubella IgG            0.00-4.9 IU/ml            ≥10.00 IU/ml 5.00-9.9 IU/ml
                                                                         6.00-15.00 AU/ml
           CMV IgG                <6.0 AU/ml                ≥6.0 AU/ml   (low Level)
           SyphilisTP
           Ab                     <1.00 S/CO                 ≥1.00 S/CO
           HBsAg
           Qualitative            <1.00 S/CO                ≥ 1.00 S/CO

           CMV IgG: 6.00-15.00 AU/ml reported as Low level.
            Rubella IgG:10.00-15.00 IU/ml reported as Low level.

VI.       Monthly Maintenance:

               a. 6012 Air Filter Cleaning, follow instruction on line.
               b. 6014 Pipettor Probe Cleaning, follow instruction on line.
               c. 6015 WZ Probe Cleaning manual, follow instruction on line.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 11
                                                                          Policy # MI/SER/16/v02                      Page 6 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

VII.      Quarterly Maintenance:

               Change probes.


VIII.      Reporting :

Specimen with value less then the cut-off value will be reported as negative. Specimen with value greater
than the cut-off value will be reported as positive. See chart below for reflex testing,and LIS reporting .


 Architect                        AXSYM                                     HBsAg                          8HBC
                                                                                                           Run reflex and
 HbsAg <2.00 IU/ml                >10.00 S/CO                               verify result value            release result
                                                                            Type in negative,
                                                                            do not release
                                  <2.00-neg                                 result value                   Delete reflex
                                                                            Do confirmation
                                                                            on AXSYM, verify
                                                                            result value)if
                                                                            confirmed, report
                                                                            as ‗Negative‘, do
                                                                            not release result             Run reflex and
                                  >2.00-<10.00 S/CO                         value.                         release result
                                                                            Do confirmation
 HbsAg >2.00 IU/ml                                                          on AXSYM                       HbcAb-negative




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 12
                                                                          Policy # MI/SER/16/v02                      Page 7 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

                                   Architect                                                               LIS
                                   HBCII                                           8HBC                          8HBC2
                                   Delete HBCII reflex from
                                   pending order. Print and release                enter as 'not done,
 HBsAg previous pos                pos result.                                     Hag +'                        N/A
                                   >5.0 S/CO, delete HBCII reflex
 HBsAg new                         from pending order. Print and                                                 enter as 'not
 Pos/HBCII ordered                 release pos result.                              Positive                     done, Hag +'
                                   >5.0 S/CO, delete HBCII reflex
                                   from pending order. Print and                                                 enter as 'not
 Only HBC ordered                  release pos result.                              Positive                     needed'
                                   HBCII +(<5.0 S/CO)-2 HBCII
                                   appear in pending order-if 2/3
                                   & 3/3 result are +,print LIST
                                   report and release 1st pos and                                                 Take off keypad
                                   delete the reflexes -do not                                                   and enter as
 HBsAg new Pos                     release the reflexes.                            Positive                     Positive
                                   HBCII +(<5.0 S/CO)-2 HBCII
                                   appear in pending order-if 2/3
                                   & 3/3 result are neg, print LIST
                                   report and release one of the
 HBsAg neg or only                 negative and delete the original
 HBC ordered                       pos and one of the neg reflexes.                Negative                      N/A
                                   HBCII +(<5.0 S/CO)-2 HBCII
                                   appear in pending order-if 2/3
                                   & 3/3 result are +,print LIST
                                   report and release 1st pos and                                                 Take off keypad
 HBsAg neg or only                 delete the reflexes -do not                                                   and enter as
 HBC ordered                       release the reflexes.                            Positive                     Positive




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 13
                                                                          Policy # MI/SER/16/v02                      Page 8 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

                                  Architect                                                                LIS
                                  HBCII                                           8HBC                           8HBC2
                                  Delete HBCII reflex from
                                  pending order. Print and release                Enter as ‗not done,
 HbsAg previous pos               pos result.                                     Hag +‘                         N/A
                                  >5.0 S/CO, delete HBCII reflex
 HbsAg new                        from pending order. Print and                                                  enter as ‗not
 Pos/HBCII ordered                release pos result.                              Positive                      done, Hag +‘
                                  >5.0 S/CO, delete HBCII reflex
                                  from pending order. Print and                                                  enter as ‗not
 Only HBC ordered                 release pos result.                              Positive                      needed‘
                                  HBCII +(<5.0 S/CO)-2 HBCII
                                  appear in pending order-if 2/3
                                  & 3/3 result are +,print LIST
                                  report and release 1st pos and                                                  Take off keypad
                                  delete the reflexes –do not                                                    and enter as
 HbsAg new Pos                    release the reflexes.                            Positive                      Positive
                                  HBCII +(<5.0 S/CO)-2 HBCII
                                  appear in pending order-if 2/3
                                  & 3/3 result are neg, print LIST
                                  report and release one of the
 HbsAg neg or only                negative and delete the original
 HBC ordered                      pos and one of the neg reflexes.                Negative                       N/A
                                  HBCII +(<5.0 S/CO)-2 HBCII
                                  appear in pending order-if 2/3
                                  & 3/3 result are +,print LIST
                                  report and release 1st pos and                                                  Take off keypad
 HbsAg neg or only                delete the reflexes –do not                                                    and enter as
 HBC ordered                      release the reflexes.                            Positive                      Positive




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 14
                                                                          Policy # MI/SER/16/v02                      Page 9 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

                                  Architect                                                                LIS
                                  HCV                                              8HCA                       8HCA2
                                  1reflex in pending order, if
                                  also>10 S/CO, release 1st pos
                                  result. Print LIST report and
 Pos >10.00 S/CO                  delete reflex HCV.                               Positive                     Enter as Positive
                                  1reflex from pending order‘, if
                                  result is <10 S/CO or becomes
                                  <1.00,print LIST report and
 Pos <10.00 S/CO                  delete reflex HCV.                               Send to PHL                  Enter as Positive

                                  If is previous HCV AB +, delete
                                  reflex order from pending list.                                               Enter as prev
 Pos >10.00 S/CO                  Report HCV as prev conf.        Positive                                      conf.
                                  If is previous HCV AB +, delete
                                  reflex order from pending list.                                               Enter as prev
 Pos <10.00 S/CO                  Report HCV as prev conf.                         Positive                     conf.
                                  HbeAg & Anti-Hbe
                                  2 reflexes in pending order. If
                                  2/3 neg ,print LIST report and
                                  release one of the negative and
                                  delete the two remaining result.
                                  If 2/3 pos, print LIST report and               Enter
                                  release one of the positive and                 positive/Negative
                                  delete the two remaining                        depending on 2/3
 0.9-1.1S/CO                      results.                                        results.                      N/A
                                  Rubella                                         8RUB                          8RUB2
                                  1 reflex in pending order, if
 0.0-9.9 IU/mL                    both neg.                                       Negative                      Negative
                                  1 reflex in pending order, if is                Positive @ low
                                  >10.0                                           level                         positive
                                  CMV IgG
                                                                                  Enter as Positive
                                                                                  @ low value
 Gray Zone                        6-15 Au/ml                                                                    N/A




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 15
                                                                          Policy # MI/SER/16/v02                      Page 10 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

    Verification:
    1. Verify all previous POSITIVE HbsAg & HCV Ab results
    2. HBCII + (>5.0) or (<5.0)2/3 +/-,initial and date on printout form.
    3. HbeAg/Anti Hbe +, or 2/3 neg, initial and date on printout form.
    4. Rubella Neg/Gray zone.
    5. CMV gray zone (6-15 Au/ml).
    6. Syphilis TP Ab is not to be used for cadaveric specimens. Send all Positive Syphilis
    screen to PHL for confirmation.

IX.       Quality Controls:

          Abbott Controls:
          Positive and negative controls must be run for all assays once per 24 hour shift. Repeat with a
          fresh aliquot if result is 1-3s in Levy-Jennings graph. If still 1-3s upon repeating, do calibration
          .First control value outside 2sd is a warning,but if this happened two days in a row.2nd day repeat
          with new aliquot,if still outside 2 sd upon repeating,do calibration.
          If 2 controls are 2-sd, do calibration.
.
          Control results are verified by a senior technologist and filed in AxSYM printout log.

          External Controls:
          Run Positive External Control and Negative External Controlswith each new reagent lot and when
          regular QC problems
          Virotrol® I Class F is for :HBsAg, HIV1&2Ab, HBcore Ab, HCV Ab,
          Virotrol® III class A is for HBeAg, HbcIgM, and HAVIgM
          Virotrol® II class A is for Anti-HBs, and Anti-HAVIgG
          Virotrol® IV is for Anti-HBe
          Virotrol® Torch is for CMV IgG and Rubella IgG
          Accurun 156 is for Syphilis TP Ab.
          Viroclear® is negative for HBsAg, Anti-HBs, HIV1&2Ab, HBcore Ab, HbcIgM, HCV Ab,
          HBeAg, Anti-HBe, HAVIgM and HAVIgG
          Rubella Negative used is a pooled Negative Proficiency testing samples(e.g CAP).

          All stock Accuruns/ Virotrols® are stored at –700C freezer (MIFTJ).

          External Controls are entered by technologist in & Architect Reagent Log binder and Architect
          External control spreadsheet. All QC controls are verified by a senior technologist.

          If any external control result is out of range, withhold test results and consult with Charge/senior
          technologists for review.

          CAP, QMP-LS, and NML provide external proficiency testing

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 16
                                                                          Policy # MI/SER/16/v02                      Page 11 of 11
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Architect System

          Calibrations:
          Run calibration for each new reagent lot or when control(s) fail.

          Failed QC:
          Test is invalid without satisfactory QC results. Do not release reagent for use pending resolution


X.        Reference

          Abbott Operation Manual (201837-106).
          Assay Packages inserts.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 17
                                                                          Policy # MI/SER/04/v09                      Page 1 of 15
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: January 10, 2011
                                                                          Annual Review Date: January 10, 2011

        AxSYM System for HBsAg, HBsAb, HBcAbTotal, HCVAb, CMV IgG, and HIV-1/2

I.        Introduction

         The Abbott AxSYM System is a fully automated random access analyser that utilizes an enzyme-
         linked immunoreacting substance (antibody, antigen or hapten) adsorbed onto microparticles to
         detect the primary interaction of antibody or antigen. The reaction mixture is transferred to an
         inert glass fibre matrix to which the microparticles bind irreversibly. The immune complex on the
         glass fibre matrix is detected by alkaline phosphatase-labelled conjugate.
         AXSYM is used only for TGLN (Trillium Gift of Life) specimens, since these tests are approved
         for donor testing.

II.       Specimen Collection and Processing

          Blood is collected (5 mL ) in a serum separator tube, Sodium heparin tube, Sodium citrate tube,
          ACD tube, CPDA-1 tube or EDTA tube. Sample is centrifuged (3,000 g x 10 minutes) and the
          serum is stored refrigerated until testing. After testing is completed, specimens are stored at -700C
          for 10 years for bone marrow and bone bank patients and 3 months for other patients.
          Re-centrifugation is required for samples that are cloudy.

III.      Procedure

          Reagents

          Reagents, calibrator and control packs for the assays are stored refrigerated. All other reagents are
          stored at room temperature, except MUP (Solution 1) which is stored refrigerated. Once MUP
          solution is opened, it is only good for 2 weeks.

          AxSYM Operation (*Refer to Abbott Operation Manual vol. 1&2 for specific maintenance
          procedures)

[A]       Daily Maintenance*
          1) Flush Bulk Solution 4 for Sampling syringe, Bulk Solution 4 for Processing syringe, Bulk
             Solution 1 and 3.
          2) Initial the maintenance log book.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 18
                                                                          Policy # MI/SER/04/v09                      Page 2 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

[B]       Assay Procedure for Patient Sample (Primary tubes)

          1)        Check that label on tube contains ordered tests as per the requisition and the name on the
                    label matched the name on the tube. The bar code label is centred and vertical on the tube.
          2)        Remove the cap. Check for and remove any bubbles in the serum. Ensure that the level
                    of serum is adequate.
          3)        Load the samples and place the segments in the sample carousel.
          4)        Load the appropriate assay reagents in the reagent carousel. (Remember to open reagent
                    bottle #4 on the reagent pack, if present).

[C]       Assay Procedure for Patient Sample (Sample cups)

          1)        From the MAIN MENU, select "ORDERLIST", select F6 ―Patient‖.
          2)        Scan LIS #, type in ―Name‖, and then press ―TAB‖.
          3)        Type S/P location of the sample cup.
          4)        Select F6 - "ADD", continue the above steps for the next sample cup.
          5)        Select F1 - "EXIT" once all the patients have been entered.
          6)        Print the displayed orderlist using the PRINT KEY. Verify sample cups are in correct
                    location.
          7)        Load the samples.
          8)        Check for bubbles in the sample and place the segments in the sample carousel.
          9)        Load the appropriate assay reagents in reagent carousel. (Remember to open reagent
                    bottle #4 on the reagent pack, if present).
          10)       PRESS "RUN".

[D]       Assay Procedure for Control Sample

          1)        From the MAIN MENU, select "ORDERLIST".
          2)        Select F5 - control.
          3)        Select the assay for the control by touchscreen.
          4)        Type in the location of the control sample S/P.
          5)        Select positive/or negative control by touchscreen.
          6)        Select F6 - "ADD". Continue for next control follow.
          7)        Select F1 - "EXIT" Repeat control for next assay or Press "ABBOTT LOGO "TO "MAIN
                    MENU".
          8)        Review the displayed orderlist. To print the orderlist, select the PRINT KEY.
          9)        Load the control samples.
          10)       Check for bubbles in the sample and place the segments in the sample carousel.
          11)       Load the appropriate assay reagents in reagent carousel. (Remember to open reagent
                    bottle #4 on the reagent packs, if present).
          12)       Press "RUN".


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 19
                                                                          Policy # MI/SER/04/v09                      Page 3 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

                    INDEX CALIBRATION: HBsAg, HCVAb, HIV-1/2

                    Index Calibration procedure requires only one sample cup. Check required volume on
                    printed Orderlist.

                    1) Order Cal from the MAIN MENU, select "ORDERLIST".
                    2) Select F4 - "CAL".
                    3) Select the assay to be calibrated using the touch screen.
                    4) The S/P is automatically assigned. Change the S/P location if necessary.
                    5) Enter reagent lot number and expired date (found on reagent box).
                    6) Press F6 - "ADD".
                    7) Press Abbott logo (a) if no other assay calibration is required.
                    8) Print the displayed orderlist using the "PRINT KEY".
                    9) Mix the calibrator and place appropriate volume in the sample cup.
                    10) Check for bubbles in the sample and place the segments in the sample carousel.
                    11) Load the appropriate assay reagents in reagent carousel. (Remember to open reagent
                        bottle #4 on the reagent pack, if present).
                    12) Press "RUN"

                    STANDARD CALIBRATION: CMV IgG Ab

                    Master calibration requires 6 calibrators in separate S/P locations. Check required volume
                    on printed orderlist.

                    1)  Go to the MAIN MENU, select "ORDERLIST".
                    2)  Select F4 - Cal.
                    3)  Select the assay to be calibrated by using the touchscreen.
                    4)  The S/P for two master cal samples is automatically assigned. Change the S/P
                        location if necessary.
                    5) Enter master cal lot number and expiry date (number on box).
                    6) Make sure calibration type is Master Cal, if not enter "MASTER CAL" by
                        touchscreen.
                    7) Select F6 - "ADD".
                    8) Select F1 "ABBOTT LOGO" if no further request is needed.
                    9) Print the displayed orderlist using the "PRINT KEY".
                    10) Dispense calibrators into 6 sample cups.
                    11) Check for bubbles in the samples and place the segments in the sample carousel.
                    12) Load the appropriate assay reagent in the reagent carousel. (Remember to open
                        reagent bottle #4 on the reagent pack, if present).
                    13) Press "RUN".



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 20
                                                                          Policy # MI/SER/04/v09                      Page 4 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System


[E]       Daily Clean-up Procedure

          1)        Ensure that all reagents have been removed and scanned.
          2)        Clean probe*.

[F]       Monthly Maintenance*

          1)        MEIA & FPIA Verification
          2)        Clean sample segments and sample cup adaptors.
          3)        Clean matrix cell & processing carousels.
          4)        Clean air filters.
          5)        Clean outside of probes
          6)        Clean wash cups/waste stations.
          7)        Clean dispenser nozzles and baseplate.
          8)        Flush pumps and syringes.
          9)        Weekly Archive patient results (must be performed before 1,500 records).
          10)       Tubing decontamination.

Maintenance Procedure:
Daily Maintenance:

       a. Bulk Solutions:
          i.     Open Waste and Supply Center Doors:
                 1. Remove the bulk solution that you are replacing.
                 2. Load the replacement solution bottle and screw on the cap.
                 3. Close the Waste and Supply Center Doors.
          ii.    Update inventory:
                 1. Select Inventory from Main Menu.
                 2. Select F5- Bulk Solutions
                 3. Select F2- Replace Solution 1
                           F4- Replace Solution 3
                           F5- Replace Solution 4
                 4. F6- Save, Exit to Main Menu.

1.    Flushing Pumps and Syringes:
      Select Maintenance from the Main Menu.
      Select Prime and Flush.
      Enter 1 under sampling syringe flush and Processing syringe flush.
      Select F2- Perform Maint.
      Check for leaks or bubbles in the tubings.
      Exit to Main Menu.

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 21
                                                                          Policy # MI/SER/04/v09                      Page 5 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

2. After loading all reagent packs, press F5-Scan pack,
   Press ‗Inventory‘.
   Press F3 - ‗Reagent Load List‘. Check to see how many tests are left in each reagent pack.
   To print a copy, Press ‗Alt‘ and ‗PRINT‘ at the same time.

3. Scan reagent packs at the end of the day:
    a. Remove all Reagent Packs.
    b. Select F5- scan pack from Main Menu.

4. Probe Clean at the end of the day :
     a. Select Maintenance from the Main Menu.
     b. Select Probe Clean. Follow instructions on the screen.
     c. Load two Reaction Cells at the designated locations on the Reaction Vessel Carousel.
     d. Pipette 1 ml of TEAH (Probe Clean Solution) into both the buffer and predilution wells of
        both Reaction Vessels.
     e. Select OK.
     f. When finished, exit to Main Menu.

5. Archive new results of the week on Friday.

Monthly Maintenance:

1. Replace and cleaning Air Heater Inlet Filter.
2. Replace and cleaning Card Cage Air Filter.
3. Cleaning Sampling Probe.
4. Cleaning Processing Probe.
5. Cleaning Sampling Wash Station.
6. Cleaning Processing Wash Station.
7. Cleaning Dispenser Nozzles.
8. Flushing Pumps and Syringes.
9. Cleaning Processing Carousel.
10. Cleaning Matrix Cell Carousel.
11. MEIA Verification.
12. FPIA Verification.
13. Cleaning Segments and Sample Cup Adapters.

Remove Cell Hopper, and lift Processing Center Cover.

1.     Replace and Cleaning Air Heater Inlet Filter:
       The Air Heater Inlet Air Filter is located on the left hand side and near the back of the AXSYM.
       Lift filter up and replace with a clean filter in the filter slot.
       Clean the dirty filter under running tap water in the opposite direction of the airflow arrows.
       Shake off excess water and air-dried.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 22
                                                                          Policy # MI/SER/04/v09                      Page 6 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

2.     Replace and Cleaning Card Cage Air Filter
       Open the Card Cage Access Panel located on the left side of the system.
       Remove the Card Cage Filter.
       Place the spare filter in the filter slot.
       Install the Card Cage Access Panel.
       Clean the dirty filter under running tap water in the opposite direction of the airflow arrows.
       Shake off excess water and air-dried.

       Select Maintenance from Main Menu.
       Select PRIME AND FLUSH.
       Select F6- Raise probes.
       Open the sampling Pipette Cover.

3.     Cleaning Sampling Probe
       Moisten a cotton swab with Deionized H2O.
       Wipe the Sampling probe several times.
       Moisten a cotton swab with 95 Ethanol.
       Wipe the Sampling probe several times.
       Moisten a cotton swab with Deionized H2O.
       Wipe the Sampling probe several times.
       Visually check the probe for damage.

4.     Cleaning Processing Probe:
       Moisten a cotton swab with Deionized H2O.
       Wipe the Processing probe several times.
       Moisten a cotton swab with 95 Ethanol.
       Wipe the Processing probe several times.
       Moisten a cotton swab with Deionized H2O.
       Wipe the Processing probe several times.
       Visually check the probe for damage.

5.     Cleaning Sampling Wash Station:
       Remove the Wash Station.
       Rinse the Wash Station with Deionized H2O.
       Rinse the Wash Station with 95 Ethanol.
       Rinse the Wash Station with Deionized H2O.
       Clean the inside and outside of the Wash Station with a cotton swab moistened with Deionized
       H2O to remove any salt buildup.
       Reinstall the Wash Station.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 23
                                                                          Policy # MI/SER/04/v09                      Page 7 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

6.     Cleaning Processing Wash Station:
       Remove the Wash Station.
       Rinse the Wash Station with Deionized H2O.
       Rinse the Wash Station with 95 Ethanol.
       Rinse the Wash Station with Deionized H2O.
       Clean the inside and outside of the Wash Station with a cotton swab moistened with Deionized
       H2O to remove any salt buildup.
       Reinstall the Wash Station.
       Home Probes.

7.    Cleaning Dispenser Nozzles:
      Remove the Bulk Solution 1 Dispenser by pulling olive-green lever toward you (↓).
      Lift the Dispenser to inspect the nozzle for buffer deposits.
      Moisten a cotton swab with Deionized H2O and clean the outside of the nozzle and the
      surrounding collar.
      Inspect the nozzle for obstruction.
      Reinstall the Dispenser by lifting the black clip to vertical position.( should hear ‗click‘).
      Check the Dispenser position. Make sure that the Dispenser sits properly, not on a raised
      position.

       Remove the Bulk Solution 3 Dispenser by Dispenser by pulling olive-green level toward you (↓).
       Lift the Dispenser to inspect the nozzle for buffer deposits.
       Moisten a cotton swab with Deionized H2O and clean the outside of the nozzle and the
       surrounding collar.
       Inspect the nozzle for obstruction.
       Reinstall the Dispenser by sliding the metal hinge pin on the right end of the Dispenser into the
       openings on the base. Lower the left end into the circular opening of the base plate.
       Check the Dispenser position. Make sure that the Dispenser sits properly, not on a raised
       position.

8.     Flushing Pumps and Syringes:
       Select Prime and Flush
       Enter 1 under sampling syringe flush and Processing syringe flush.
       Select F2 - Perform Maint.
       Check for leaks or bubbles in the tubings.
       Exit to Main Menu.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 24
                                                                          Policy # MI/SER/04/v09                      Page 8 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

9.    Cleaning Processing Carousel:
      Remove the Processing Carousel Cover by unscrewing the two thumbscrews.
      Grasp Processing Carousel Motor Handle, push back and hold open.
      Remove by tilting Carousel and pulling up, careful not to hit the probe.
      Clean the carousel and the surrounding area with lint-free tissue moistened with water.
      Inspect the clips on the under side of the carousel for broken pieces/loose connections.
      Dry the Carousel with Lint free tissue.
      Reinstall the Carousel by grasping the Processing Carousel Motor Handle, push back and hold
      open.
      Place the rim of the Carousel in the V-wheel located under the Carousel Motor Housing.
      Tilt the Carousel downward and fit the rim into the two front V-wheels. Careful not to hit the
      probe.
      Reinstall the Processing Carousel Cover by tightening the two thumbscrews.

10. Cleaning the Matrix Carousel:
    Remove the Matrix Cell Carousel Access Panel.
    Remove the Air Deflector.
    Select F2- Shutdown from the Main Menu.
    Verify that Shutdown was selected, select OK.
    Turn the power switch to OFF. Once power is off, make sure that it is off for at least 5 minutes
    before turning AXSYM back ‗ON‘.
    Rotate the Carousel so that the red dot is facing you.
    Remove the Carousel by pushing back on the Carousel, tilting down and pulling out.
    Clean the carousel and the surrounding area with lint-free tissue moistened with water.
    Inspect the clips on the under side of the carousel for broken pieces/loose connections.
    Dry the Carousel with Lint free tissue.
    Reinstall the Carousel by holding the carousel with the red dot facing you, and fit the rim of the
    carousel into the rear V-wheel and push in towards the spring-loaded V-wheel.
    Tilt up and fit the rim of the carousel into the front V-wheels.
    Manually rotate Carousel to ensure proper positioning.
    Reinstall Air Deflector. Reinstall the Matrix Cell Carousel Access Panel.
    Close the Processing Center Cover.
    Re-install Matrix Cell Hopper.
    Turn the power ON.
    Select Startup from the Main Menu.
    If the power is off for less than 15 minutes, there is a 15-minute warm-up.
    If the power is off for more than 15 minutes, there is an hour warm-up.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 25
                                                                          Policy # MI/SER/04/v09                      Page 9 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

11. MEIA Verification:
    Select MAINTENANCE from the Main Menu.
    Select CALS AND CHECKS
    Select OPTICS.
    Select MEIA VERIFICATION.
    Remove Matrix Cell Hopper.
    Select CONTINUE. Follow instructions on screen.
    Place the Standard Hopper into position.
    Insert the Standard into the Standard Hopper.
    Select CONTINUE to begin the MEIA Verification.
    This will take  10 minutes.
    When finished, select OK to acknowledge completion of the verification.
    Press PRINT on the keyboard.
    Retrieve the Standard by pressing the white MEIA Standard Release lever.
    Open the MEIA Optical Retrieval Door and remove the Standard.
    Store it upside down in the black box.
    Reinstall Matrix Cell Hopper.
    Exit to the MAIN MENU/MAINTENANCE/OPTICS.

12. FPIA Verification:
    Select FPIA VERIFICATION.
    Follow instruction on the screen, select Continue.
    Press down the back of the clip on top of the FPIA Optical Standard.
    Load the Standard into the Reaction Vessel Carousel at the Load Station.
    Press on the front of the clip to provide a flat surface.
    Select Continue to begin the procedure.
    This will take  20 minutes.
    When finished, select OK to acknowledge completion of the verification.
    Press PRINT on the keyboard.
    Press down on the back of the clip on top of the FPIA Optical Standard.
    Lift and remove the Standard, store in assigned place in the black box.
    Exit to Main Menu.

13. Cleaning Segments and Sample Cup Adapters:
    Use disinfectant wipes to clean segment and Sample Cup Adapters.
   Empty Waste Containers first thing in the morning or at the end of the day.
   Open the Waste and Supply Center Doors ( DO NOT OPEN THIS DOOR WHEN THE AXSYM
   IS RUNNING ) , then open the Interior Waste Door :
   a. Remove Consumable waste
     i.      Remove biohazard bag from the consumable waste container and dispose it into
             Biohazard waste box.
     ii.     Place a new biohazard bag in the container.

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 26
                                                                          Policy # MI/SER/04/v09                      Page 10 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

     b. Remove Liquid Waste
       i.     Hold down the tubing to the Liquid Waste Container while pressing the
              metal tab on the side of the clip.
       ii.    Remove the Liquid Waste Container.
       iii.   Transfer liquid waste into decontamination containers near the sinks. Fill container with
              4/5 full of waste and 1/5 full of Javex. Label date on the lid.
       iv.    Rinse container with a little bit of Javex, and then lots of H2O. Wipe dry outside of
              container.
       v.     Reattach the tubing by pressing the tubing assembly until it locks into place.
      Close the Interior Waste Door, and close the Waste and Supply Center Doors.

1. Update Waste Levels:
    a. Select Inventory from the Main Menu.
    b. Select F4- Waste.
    c. Select F3- Empty Waste.
    d. Select F6- Save.
    e. Exit to Main Menu.

2.    Update Matrix cells and Bulk Solutions:
       a. Matrix Cells:
          i.     Visually check inventory level.
          ii.    If level is around or less than 100, add a box by removing the plastic cover, and the
                 paper tabs in the middle of the box. Place the box between the plastic walls at the top
                 of the Matrix Cell Hopper. Rest the box ends firmly on the roll bars and apply
                 pressure to the center of the box. The Matrix cells pour into the Hopper. Make sure
                 that no Matrix
                 Cells lie sideways.
          iii.   Update inventory:
                 1. Select Inventory in Main Menu.
                 2. Select F2- Matrix cell.
                 3. Select F3- Add a box.
                 4. Select F6-Save.
                 5. Exit to Main Menu.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 27
                                                                          Policy # MI/SER/04/v09                      Page 11 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

IV.       Results

          Results should be sorted and viewed by S/P to ensure that all tests were completed then resorted
          and printed.
          1)      From the MAIN MENU, select "RESULTS".
          2)      Select F5 - SORT.
          3)      Select "S/P' by touchscreen, "OK".
          4)      Scroll down the list checking that every S/P with a sample has a result.
          5)      Select F5-SORT.
          6)      Select "SID" OK. Once sorted, select F2-Select All and PRINT.
          7)      Select "Print a listing of all selected results", "OK" using touchscreen to activate printing.
          8)      After successful printing of results, select F3 "Release Results". "Are you sure? - Yes".
          9)      Check print-out for low level positive CMV, Rubella and enter the low level message in
                  the LIS. Check for reactive HBsAg, HBcAb, HCVAb, HIV 1/2, for which repeat testing
                  or reflex testing may be necessary.
          10)     Initial and give print-out to senior technologist for verifying.

          Note:
                    HBcAb:               Repeat all reactive results for HBcAb. After repeat testing complete.
                                         Select the appropriate result for 8HBC test from keypad in LIS.
                                         (Exception: HBcAb reflex testing (8HBC2) performed on HBsAg reactive
                                         sample for which HBcAb (8HBC) was not originally ordered as outlined
                                         below.

                    HBsAg:               Perform reflex confirmatory testing and HBcAb on all reactive HBsAg
                                         samples except for patients with previously confirmed positive HBsAg.
                                         Preliminary report of stat requests will be phoned as "presumptive
                                         positive".




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 28
                                                                          Policy # MI/SER/04/v09                      Page 12 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

 V. Reporting Results

       i. HBsAg Assay Cut-off: >2.0 S/N

                    Reactive and confirmed, HBcAb reactive:     POSITIVE*
                    Reactive and previously confirmed positive: POSITIVE
                    Reactive and non-confirmed: Negative

                              HBsAg                              HBcAb                               Action Required
                                S/N                               Result
                              2                                  + or -                    Report: Negative
                              2-5                                   +                       Send to PHL
                              >5 -10                                +                       Perform Confirmatory Testing
                               2 – 10                               -                       Send to PHL
                                >10                                 -                       Perform Confirmatory Testing
                                >10                            + S/CO <0.2                  Report: Positive
                                >10                            + S/CO >0.2                  Perform Confirmatory Testing


          *Report new cases to Medical Officer of Health (alert senior/charge technologists by underlining
          AxSYM print-out in red, senior/charge will send copy to report controller for MOH notification)

       ii. HBsAb Assay Cut-off: <1.0 S/CO

                    Repeatable1 reactive:                     POSITIVE
                    Negative:                                 Negative
                    Non-repeatable1 reactive:                 Negative
                    1
                     Centrifuge sample at 20,000g for 10 minutes and re-check patient label before repeating,
                    document on AxSYM print out and initial

     iii. HCV Ab Assay Cut-off: >1.0 S/CO

                    1.         If S/CO > 10, reflex to 8HCA2. Report POSITIVE* if repeatably1 reactive.
                    2.         If S/CO < 10, enter in the LIS as "To PHL @ MOH+". Send to PHL next day.
                               Preliminary report of STAT testing will be phoned as "Presumptive Positive".

                               Reactive:     POSITIVE*
                               Non-reactive: Negative
                               Repeatedly Reactive cadaver samples S/CO 1.0 - 10.0:
                               Indeterminate@CAD1


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 29
                                                                          Policy # MI/SER/04/v09                      Page 13 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System


                    *Report new cases to Medical Officer of Health (alert senior/charge technologists by
                    underlining AxSYM print-out in red, senior/charge will send copy to report controller for
                    MOH notification)
                    1
                      Centrifuge sample at 20,000g for 10 minutes and re-check patient label before repeating,
                    document on AxSYM print out and initial.


                               Repeatedly Reactive cadaver samples S/CO 1.0 - 10.0:
                               Indeterminate@CAD1


                    *Report new cases to Medical Officer of Health (alert senior/charge technologists by
                    underlining AxSYM print-out in red, senior/charge will send copy to report controller for
                    MOH notification)
                    1
                      Centrifuge sample at 20,000g for 10 minutes and re-check patient label before repeating,
                    document on AxSYM print out and initial.

     iv. CMV IgG Antibody Cut-off: >15 AU/mL

                    Positive (>25 AU/ml) :                     POSITIVE
                    Positive (15-25 AU/mL):                    POSITIVE (Low Level)
                    Negative (< 15 AU/ml):                     Negative

     v. HIV 1/2 Assay
                    ***Reactive: Enter in the LIS as "TO PHL @MOH+". Send to PHL next day for
                                 confirmation.
                         Negative: Negative




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 30
                                                                          Policy # MI/SER/04/v09                      Page 14 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System

VI.       Quality Control
         Abbott Controls:
         Positive and negative controls must be run for the following assays once per 8 hour shift: HCV
         Ab, and CMV Ab .
         Positive and negative controls must be run for the following assays once per 24 hour shift:
         HBsAg, HBc Ab and HIV1&2Ab
.
         Results are verified by a senior technologist and filed in AxSYM printout log.
         External Controls:
         Run Positive External Control with each new reagent lot and with QC problems
         Accurun 5100 is for :HBsAg, HIV1&2Ab, HBcore Ab, HCV Ab, and CMV IgG

         Viroclear is used as negative control for all of the above tests.

          All stock Accuruns are stored at –700C freezer (MIFTJ).

          External Controls are verified by a senior technologist and filed in External QC & Reagent Log
          Chart binder,also enter in spreadsheet in T:/Microbiology/Virology/QC Statistic/AXSYM.

          If any external control result is out of range, withhold test results and consult with Charge/senior
          technologists for review.

         CAP, QMP-LS, and LCDC provide external proficiency testing

          Calibrations:
          Run calibration for each new reagent lot or when control(s) fail.

          Failed QC:

          Test is invalid without satisfactory QC results.

               a. Do not release reagent lot for use pending resolution of QC error.
               b. Inform charge/senior technologist.
               c. Record in Reagent Log Chart, Instrument Maintenance Log if microscope/incubator is
                  involved in the failure (and Incident Report if necessary).
               d. Re-run failed control materials in parallel to fresh controls to evaluate the QC material
                  itself.
               e. If the re-run shows the old QC material still fails and fresh QC is satisfactory, the error
                  may be attributed to the old QC material itself and the reagent is satisfactory.
               f. If the re-run shows both the old and fresh QC materials fail (and other QCs not
                  satisfactory), the error may be attributed to the reagent then the reagent cannot be
                  released for use.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 31
                                                                          Policy # MI/SER/04/v09                      Page 15 of 15
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM System


VII.      Reference

          AXSYM operational manual and manufacturer's package insert for the appropriate assay.

          (Abbott Laboratories, Diagnostics Division, Abbott Park IL 60064, USA)




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 32
                                                                          Policy # MI/SER/05/v05                          Page 1 of 4
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM HBsAg Surface
                                                                                         Antigen Confirmatory Assay
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: September 20, 2001
                                                                          Annual Review Date: January 10, 2011

                                     AxSYM Heptitis B Surface Antigen Confirmatory Assay

I.        Introduction

          AXSYM HBsAg Confirmatory assay is a Microparticle Enzyme Immunoassay (MEIA) used for
          the detection of HBsAg in human serum or plasma by means of specific antibody neutralization.

II.       Specimen Collection and Processing

          A minimum of 300 L of preliminary positive HBs Ag patient sample in a sample cup is required.

III.      Procedure

          i)        Reagents

                    AxSYM Confirmatory Kit
                    AxSYM HBsAg Kit

          ii)       Method

                    Use the assigned "H" segment for the confirmatory test.

                    Using the following chart for the number of samples to be confirmed, add appropriate
                    volume of reagents to the corresponding sample cups at position 1 to position 5. Patient
                    sample starts at position 6 onward.

                    Segment "H"

                    S/P        1         =          Reagent A

                    S/P        2         =          Reagent B

                    S/P        3         =          Dilution Reagent

                    S/P        4         =          Index Calibrator 9use cal. Received with reagent kit)




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 33
                                                                           Policy # MI/SER/05/v01                         Page 2 of 4
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                   Subject Title: AxSYM HBsAg Surface
                                                                                          Antigen Confirmatory Assay

                    S/P        5         =          HBs Ag Positive Control

                    S/P        6         =          Patient sample


Sample                Reagent                                     For Cal/          Minimum Volume Required (L)
Segment                 or                     Reagent            Control                No. of Patient Samples
Position              Sample                    Color              Only             1     2         3      4    5

     1              Reagent A                   Violet               100          150         200         250        300         350

     2              Reagent B                  Yellow                150          200         250         300        350         400

     3                Dilution                 Natural                0           500         875        1250        1625       2000
                      Reagent

     4                Index                     Green                270          270         270         270        270         270
                    Calibrator*

     5               Positive                    Blue                250          250         250         250        250         250
                    Control**

     6                Patient                                                     275         275         275        275         275
                      Sample

     7                Patient                                                                 275         275        275         275
                      Sample

     8           Patient Sample                                                                           275        275         275

     9                Patient
                      Sample                                                                                         275         275

    10                Patient
                      Sample                                                                                                     275

          *270 L = 8 drops
          **250 L = 7 drops



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 34
                                                                          Policy # MI/SER/05/v01                          Page 3 of 4
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM HBsAg Surface
                                                                                         Antigen Confirmatory Assay


Index Calibrators

          1)        Select F4 - "CAL"
          2)        "HBs Ag CNF" by touch screen input
          3)        Assign S /P = H4
          4)        Enter lot number and expiry date found on confirmatory kit used.
          5)        Select F6 - "ADD"

Positive Control

          6)        Select F6 - "PATIENT"
          7)        Type "Pos Control" at SID
          8)        Assign S / P = H5
          9)        Select "HBs Ag CNF" by touch screen
          10)       Select F4 "DIL/REPS"       Order: Reagent B                              1
                                                      Reagent A                              1
                                                      Reagent B dil                          0
                                                      Reagent A dil                          0
          11)       Select F6 - "NEXT'
          12)       Select F6 "ADD"

Patient Samples
       13)   Select F6-―PATIENT‖
       14)   Barcode in SID, type in patient‘s last name
       15)   Select ―HBsAg CNF‘ by touch screen
       16)   Assign S / P H6
       17)   Select F4 - "DILS / REPS" Order: Reagent B            1
                                                    Reagent A      1
                                                    Reagent B dil 1
                                                    Reagent A dil 1
       18)   Select F6 "NEXT"
       19)   Select F6 "ADD" for more patient entry
       20)   Select "Abbott Logo" to "MAIN MENU"
       21)   Review the displayed order list. Print the orderlist using the "PRINT KEY"
       22)   Load Segment "H" containing the required confirmatory reagents and patient sample
       23)   Load HBsAg reagent in reagent carousel
       24)   PRESS "RUN"




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 35
                                                                          Policy # MI/SER/05/v01                          Page 4 of 4
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: AxSYM HBsAg Surface
                                                                                         Antigen Confirmatory Assay


IV.       Results

          1)        From the MENU, select "RESULTS"
          2)        Highlight all control and patient results associated with the confirmatory run
          3)        Select "PRINT", choose "Print a formatted result for each displayed test"
          4)        Select "OK" by touch screen


V.        Reporting Results

          HBsAg Confirmatory (8CON) testing results are entered into the LIS using the keypads but do
          not print on the report. The repeat HBcAb (8HBC2) results also do not print, but are
          automatically posted to the LIS. These confirmatory and repeat HBcAb (8HBC2) results are
          used to edit, if necessary, the HBsAg (8HAG) and HBcAb (8HBC) results which will print on
          the report.

          HBsAg Confirmatory Assay

          % Neut or % Neut Dil Positive: Confirmed Positive
          % Neut AND % Neut Dil Negative: Not Confirmed

          HBcAb (8HBC2) Assay

          Reactive:            Reactive
          Negative:            Negative

          Refer to AXSYM portion of manual for guidelines on reporting HBsAg, and HBcAb based on
          reflex testing.


VI.       Quality Control

          A positive control confirmation should be included on each run.


VII.      Reference

          AXSYM operational manual and manufacturer's package insert. (Abbott Laboratories, Abbort
          Park, IL 60064, U.S.A.)


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 36
                                                                          Policy # MI/SER/06/v02                           Page 1 of 3
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Labour & Delivery Hepatitis and
                                                                                         HIV Virus Serology
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: March 19, 2008
                                                                          Annual Review Date:

                                              Labour and Delivery Hepatitis Virus Serology

Although there are many infectious causes of hepatitis, the majority are caused by hepatitis A, B and C
viruses. Acute Hepatitis A is diagnosed by detection of IgM antibodies. Anti-HAV IgM becomes
positive just before the development of clinical hepatitis and remains positive for at least 4 months after
infection. There is no chronic carrier state for Hepatitis A. Detection of total antibodies to Hepatitis A
indicates immunity due to either past infection or immunization.

Hepatitis B is diagnosed by either detecting hepatitis B surface antigen (HBsAg) which indicates the
presence of infectious virus, (HBcAb-IgM) or Anti-hepatitis B surface antibodies (HBsAb) which
indicate immunity due to either past infection or immunization. Anti-hepatitis B core IgM antibodies
indicate acute infection and HBcAb-Total indicates previous or current infection. HBsAg should be
cleared within 6 months of acute infection. Persistence of HBsAg beyond 6 months is consistent with
chronic hepatitis B infection. Some of these tests may occasionally be performed on a STAT basis
because of concern regarding transmission of the virus to a susceptible individual following exposure (i.e.
needlestick, newborn) to infected blood/body fluids and the need to prevent the disease by administering
vaccine and/or immunoglobulin.

Hepatitis due to the delta virus is rare in Canada. It only occurs in association with patients who are
positive for hepatitis B surface antigen. Requests for this virus should be forwarded to the Public Health
Laboratory.

Hepatitis C (HCV) is a blood borne virus closely associated with blood transfusion and intravenous drug
use. The presence of antibodies to HCV indicates that an individual may have been infected with HCV,
may harbour infectious HCV and\or may be capable of transmitting HCV infection.
The following requests will be handled in our laboratory:

          Hepatitis B surface antigen                         HBsAg
          Hepatitis B surface antibody                        HBsAb
          Hepatitis B core antibody                           HBcAb
          Hepatitis B e antigen                               HBeAg1
          Hepatitis B e antibody                              HBeAb1

          Hepatitis A IgM antibody                            HAV-IgM2
          Hepatitis A Total antibody                          HAV-IgG




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 37
                                                                          Policy # MI/SER/06/v02                           Page 2 of 4
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Labour & Delivery Hepatitis and
                                                                                         HIV Virus Serology

Note: 1)            These tests will be performed on request only if the patient is HBsAg positive.
      2)            Perform only HAV-IgM if type of Hepatitis A test is not specified.

Table 1. Tests performed as per designated categories
                                                                              Test Performed
Clinical Category                                    HBsAg                   HBsAb        HAV-IgM                        HCVAb
Hepatitis B Screen                                     X
Hepatitis A Screen                                                                                      X
Hepatitis B Immune Status                                                        X
Needlestick
        - Patient (source)                               X                                                                   X
        - Staff (exposed)                                                        X                                           X
Pre/postnatal
        - Mother                                         X
Hepatitis Screen                                         X                                              X                    X

For additional requests within the above categories, consult with the charge technologist or medical
microbiologist. In the absence of one of the above clinical categories, do the tests as requested.

Table 2. Guidelines for STAT Testing

Clinical                       Serum                Time frame                     Days of              Call-
setting                        tested               from exposure                  week                 back
                                                    to report

Neonate                        Mother               < 12 hours                     All                  No1
Prenatal                       Mother               < 12 hours                     All                  No1
Needlestick/                   See below            < 72 hours                     All                  No1
mucosal exposure2
Renal dialysis                 Patient              < 12 hours                     All                  No1


     1) May require call back on weekends if time from exposure to reporting exceeds 12 hours. Must be
        approved by microbiologist on-call.
     2) Perform HBsAg and HCVAb on the source. Test HBsAb and HCVAb on the employee.
        Whichever of these arrives first is to be tested STAT. If both arrive at the same time, test both
        simultaneously. If the source HBsAg is positive, test the employee STAT. If the source is
        negative, test employee in the next routine run.

Labor Delivery Stat HIV HBsAg Instructions


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 38
                                                                          Policy # MI/SER/06/v02                         Page 3 of 3
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Labour & Delivery Hepatitis and
                                                                                         HIV Virus Serology


Labour and Delivery Human Immunodeficiency Virus (HIV) 1 & 2 Serology

HIV testing will be done on patient (source) involved in‘ Needle Stick Incident‘ as ‗STAT‘, so
appropriate prophylaxis can be given in case it is positive. It is also done for transplant patients (donor
& recipient). Two tubes of blood should be received. If the result is positive, send the unopened tube
to PHL. The positive result will be reported as ‗sent to PHL for confirmation‘. Specimen will be sent
out as ‗STAT‘ the next working day to PHL. Put specimen with PHL HIV form in a separate brown
bag, clearly marked as ‗STAT‘. Call 416-340-6022 to inform PHL HIV lab that this specimen is
coming as ‗STAT‘, and ask to call result to Lab when it is available.

Labor Delivery Stat HIV HBsAg Instructions




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 39
                                                                          Policy # MI/SER/07/v01                           Page 1 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology
Issued by: LABORATORY MANAGER                                             Original Date: November 18, 2009
Approved by: Laboratory Director                                          Revision Date:
                                                                          Annual Review Date: January 10, 2011

               HBsAg , HIV 1/2,HBcAb, HCV Ab & HTLV I/II (cadaver/donor) Serology

I.        Introduction

          Genetic Systems ™ HBsAg EIA 2.0 is the Genetic Systems Corporation qualitative enzyme
          immunoassay for the detection of Hepatitis B Surface Antigen (HBsAg) in human serum and
          plasma, and also in cadaverous serum specimens.
          Genetic Systems ™ HIV-1/HIV-2 Plus O EIA is the Genetic Systems Corporation qualitative
          enzyme immunoassay for the detection of Human Immunodeficiency Virus type 1 and /or
          Human Immunodeficiency Virus type 2 in human serum and plasma, and also in cadaverous
          serum specimens.
          Ortho Hepatitis C Version 3.0 ELISA Test System: Hepatitis C Virus Encoded Antigen
          (Recombinant c22-33, c200 and NS5) – (Donor Testing)

          Ortho Hepatitis B core ELISA Test System (Donor Testing)

          Ortho HTLV-I/HTLV-II Ab-Capture ELISA Test System (Donor Testing)
          These kits are used for postmortem/Donor specimens.
          Specimens are usually sent from the following institutions:
             a. Eye Bank of Canada (Ontario Division).
             b. Tissue Lab (Hospital For Sick Children).
             c. Skin Bank (Sunnybrook Hospital).
             d. Bone Bank (Mount Sinai Hospital).
             e. Northern Alberta EyeBank-Edmonton ProvLab, Alberta.
             f. Southern Alberta EyeBank-Calgary ProvLab, Alberta.
             g. National Capital Regional bone bank.
             h. Coroner‘s office.
             i. Referrals from PHL.


II.       Specimen Collection and Processing

               a. Blood, serum or plasma collected by Eye Banks or other clients.
               b. If blood tube received, centrifuge at 3000 rpm for 10 minutes.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 40
                                                                          Policy # MI/SER/07/v01                           Page 2 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

III.      Procedure

               EVOLIS ™ Analyzer

          i) Reagents:

               BioRad HBsAg and HIV1/2 kits:
                     1. HBsAg Conjugate Concentrate
                     2. HBsAg Conjugate Diluent
                     3. AntiHBsAg Microwell Strip Plates
                     4. HBsAg Negative Control
                     5. HBsAg Positive Control
                     6. HBsAg Low Positive Control
                     7. EIA Wash Solution (30X)
                     8. EIA Chromogen
                     9. EIA Substrate Buffer
                     10. EIA Stopping Solution
                     11. HIV 1&2 Plus O Microwell Plate
                     12. HIV 1&2 Specimen Diluent
                     13. HIV Negative Control
                     14. HIV 1 Positive Control
                     15. HIV 2 Positive Control
                     16. HIV Group O Positive Control
                     17. HIV Conjugate Concentrate
                     18. HIV Conjugate Diluent

               OrthoDiagnostic HBcAb kit:
                     1. Hepatitis B Virus Core Antigen (HBcAg) coated Microwell plates(5)
                     2. Antibody Conjugate(Murine Monoclonal)-mixture of anti-human IgG and IgM
                        conjugated to horseradish peroxidases with protein stabilizer.
                     3. Specimen diluent
                     4. OPD tablets(30) contains o-phenylenediamine∙2HCL
                     5. Substrate Buffer-citrate-phosphate buffer with 0.02% hydrogen peroxide
                     6. Positive Control(Human)
                     7. Negative control(Human)




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 41
                                                                          Policy # MI/SER/07/v01                           Page 3 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

               Orthodiagnostic HCV Ab kit:
                    1 Hepatitis C Virus(HCV) encoded Antigen coated Microwell plates (5)
                    2 Conjugate-Antibody to Human IgG(Murine Monoclonal)-anti-human IgG heavy
                        chain(murine monoclonal) conjugated to horseradish peroxidase with stabilizers
                    3 Specimen Diluent
                    4 OPD tablets(30) contains o-phenylenediamine∙2HCL
                    5 Substrate Buffer-citrate-phosphate buffer with 0.02% hydrogen peroxide
                    6 Positive Control(Human)
                    7 Negative control(Human)

                 Orthodiagnostic HTLV-I/HTLV-II Ab kit:
                     1 HTLV-I and HTLV-II recombinant antigen-coated Microwell plates (5)
                     2 Antigen Conjugate – HTLV-I and HTLV-II recombinant antigens conjugated to
                         horseradish peroxidise with protein stabilizers
                     3 Specimen Diluent
                     4 OPD tablets(30) contains o-phenylenediamine∙2HCL
                     5 Substrate Buffer-citrate-phosphate buffer with 0.02% hydrogen peroxide
                     6 HTLV-I Positive Control(Human)
                     7 HTLV – II Positive Control (Human)
                     8 Negative control(Human)

          ii) Method:

               The HBsAg , HIV 1\2, HBcore Ab, HCV Ab & HTLV-I/HTLV-II assays can be run
               simultaneously in the EVOLIS ™ analyzer.

          Before beginning EIA assay:
                   a. Fill up wash buffer containers:
                      BioRad Wash:
                      a) Prepare when necessary:
                         Add 60 ml of 30x BioRad wash solution to 1740 ml of distilled water.
                      b) Fill up Evolis wash container with blue tubing.
                       Orthodiagnostic Wash:
                       c) Prepare when necessary:
                         Add 100ml of 20X Ortho wash solution to 1900 ml of distilled water
                       d) Fill up Evolis wash container with yellow tubing
                       e) Fill up Evolis wash container with black tubing (Ortho Wash overflow)
                        De-ionized Water:
                       f) Fill up Evolis container with red tubing with de-ionized water



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 42
                                                                          Policy # MI/SER/07/v01                           Page 4 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

                        b. Transfer HBcore Ab diluent, HCV Ab Diluent and HTLV-I/II Ab Diluent into
                           separate 125 ml EVOLIS containers label and put on in-use date. The HIV
                           Diluent will be sampled from its original container.

                        c.   Transfer HBcore Ab, HCV Ab and HTLV-I/II conjugates into separate 60ml
                             EVOLIS containers, label and put on in-use-date.

                        d. Working conjugates and working TMB substrates for the HBsAg and HIV ½
                           assays are prepared fresh during each run in a 30 ml EVOLIS coated vial.

                        e. Transfer the ORTHO (4N sulfuric acid) Stop Solution into a 60ml EVOLIS
                           container and transfer the BioRad (1N sulfuric acid) Stop Solution into a 30ml
                           EVOLIS container, label and put on-in-use-date.

                        f. Aliquot HBcore Ab, HCV Ab and HTLV-I/HTLV-II Ab controls into separate
                           2ml sample containers, label and put-on-in-use-date.
                           *Use new containers for each new Lot #

                        g. HBsAg and HIV1/2 assay controls will be pipetted from their original containers
                           and can be loaded onto the analyzer as is.

                        h. Bring all microwell plates for the required assays and the above mentioned
                           reagents to room temperature before use.

                        i. Empty the waste tank and check the system liquid container (refill if necessary).
                           System Liquid Preparation: 2 ml of Tween 20 to 10L of de-ionized water.


     1 Turn ―ON‖ the EVOLIS analyzer first and then the computer at the beginning of the day.

     2 Double click on the EVOLIS icon located on the computer desk-top.

     3 Log onto the system by clicking on ―OK‖, no password is required.

     4 A self test of the system is automatically initialized each time the EVOLIS software is run. The
       self test is considered satisfactory if the word ―PASSED‖ appears beside each instrument
       module. Print a copy of the self test report and combine it with the worklist and result sheets.

     5 Prepare specimens by removing the lids and loading all sample tubes with the barcode facing
       right on to the sample rack (rack code T). Check the quality of the samples by ensuring all
       clots, foam and bubbles have been removed.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 43
                                                                          Policy # MI/SER/07/v01                           Page 5 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

     6 Open the door to the sample and reagent unit. Load the sample rack with the bar-codes facing
       the bar-code reader onto the track marked by the solid red LED light.

     7 The ‗Patient Editor‘ dialog box will appear with the specimen numbers from the loaded rack.
       Double check that the specimen numbers are correct. If there is a blank space under the patient
       ID column, click on the space and enter the specimen number manually. All specimen
       numbers entered manually will be flagged with the code ―ManID‖ on the results report.

     8 Choose the individual assays from the drop down menu which appears along the top of the
       Patient Editor dialog box, use the scroll bar at the bottom of the dialog box to select
       additional assays :
                                      HBsAg assay = GS HBsAG EIA 30
                                      HIV ½ assay = GS HIV1-2 Plus O EIA
                                     HBc_Ab assay= OR_HBC
                                    HCV_Ab assay= ACHCV_ORTHO
                               HTLV I/II_Ab assay= OR_HTLV

     9 Click on the assay box corresponding to the sample number if you want that assay to be run for
       that sample. A check mark is shown for a sample where the assay has been selected.

     10 Once all the required assays have been selected for each sample click ‗Close‘ to save. A solid
        LED light will show up for the next available track in the sample and reagent unit which
        indicates where the next sample rack can be loaded. Repeat steps 6 to 11 for each new sample
        rack that is loaded on to the analyzer.

     11 A total of four sample racks need to be loaded on to the analyzer even if all the sample racks
        are not required in order for the reagent template to be used in a later step. When an empty
        sample rack is loaded on to the analyzer just click ‗Close‘ when the Patient Editor dialog box
        appears.

     12 To check and validate the worklist click on the ‗New Worklist‘ icon located on the upper
        toolbar and the Set-Up dialog box will appear.

     13 Click on the ―+” next to each plate to see which assay is programmed for which plate. Click
        on ―+‖ beside the assay file name that is associated with each plate. The plate layout will
        appear at the right hand side of the dialog box and will show the total number of controls and
        specimens to be tested, their assigned wells and the number of strips required for each assay.
        Print plate layout.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 44
                                                                          Policy # MI/SER/07/v01                           Page 6 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

     14 If duplicate samples need to be tested for a given assay in the same run:
                   i.  From the Patient Editor dialog box select the assay folder in which the
                       duplicate sample needs to be run and click ‗Add Patient‘.
                  ii. The Select Patient(s) dialog box will appear.
                iii. Check the Allow multiple determinations box which appears at the bottom of
                       the Select Patient|(s) dialog box.
                 iv.   The patient sample IDs will appear. Click on the sample ID in which multiple
                       testing is required and click OK.
                  v.   In the Set-up Panel dialog box, a (x2) will appear beside the patient ID under
                       the assay folder in which it was selected.

     15 If all the information is correct click ‗OK‘ to validate the worklist.

     16 A separate dialog box asking for the reagent lot numbers will appear for each assay that is
        ordered. Double check the lot numbers and expiry dates and click ‗OK‘ for each dialog box
        that appears if everything is correct.

     17 Optimize the workload schedule by selecting ‗Edit‘ and ‗Optimize‘. This must be done before
        the run is started.

     18 Click on the ―+‖ to expand the Work folder and click on ‗Plate Layout‘, the number of
        microwell strips required for each assay can be reviewed. Click the ‗Print‘ button located
        along the top of the toolbar to print the plate layout if desired.

     19 Click ‗START‘ (green button) located on the upper tool bar to open the Load dialog box.

     20 The Load dialog box appears and shows where to load all the required resources.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 45
                                                                          Policy # MI/SER/07/v01                           Page 7 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

     21 Load all required resources from left to right:

                        i.     Sample Diluents
                                   HCV Diluent                 HBc Diluent
                                 HTLV I/II Diluent            HIV ½ Diluent


                         ii. Pipette tips: Grey = 1100 ul tips (full rack)
                                           Brown= 300 ul tips (full rack)
                                           Red = Incomplete tip rack
                             *The pipette tips have to be loaded in the exact position as shown in the
                             layout

                      iii.     Load the assay reagents on the reagent racks according to the following
                               templates (Figure 1-3). The reagent templates are only used when all 5 assays
                               are being run simultaneously.

                      iv.      Ensure all caps on the reagent/control containers have been removed before
                               loading them onto the analyzer.

                               Manually Assigning Reagent Locations
                               *If only individual assays are being run the location of the regents/controls have
                               to be manually assigned:
                                   a. Load the reagents and controls into the desired positions on the reagent
                                      racks and load the racks onto the analyzer. The reagent racks will appear
                                      blank when loaded.

                                    b. Different coloured circles representing the reagents/controls will be
                                       found at the right hand column of the Load dialog box under the section
                                       ‗Unallocated Resources‘. Moving the mouse over each circle tells you
                                       which reagent/control it represents.

                                    c. Click on the circle and drag it over to the correct position in which the
                                       reagent/control has been placed on the reagent rack.

                                    d. The position of bar-coded reagent/control containers do not have to be
                                       manually assigned, their location would be read by the bar code reader.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 46
                                                                          Policy # MI/SER/07/v01                           Page 8 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

Conjugates for BioRad HBsAg and HIV1/2:
      Prepare Fresh (same Day) Working Conjugate as follows (always make 1 extra strip):
                       Number of Strips 40X Conjugate (uL) Conjugate Diluent (mL)
         HBsAg (Plate A)    1               10                  1.0
                            2              20                   2.0
                            3              30                   3.0

               HIV ½(Plate B)               1                        100                           1.0
                                            2                        200                           2.0
                                            3                        300                           3.0


Chromogen(shared) for both BioRad HBsAg and HIV1/2:
     Prepare Fresh (Same Day) Working Chromogen as follows (always make 1 extra strip),
     make enough for both plates:

                                   Number of Strips Chromogen Reagent (uL)                          Chromogen Diluent (mL)
                                       1                100                                         1.0
                                       2                200                                         2.0
                                       3                300                                         3.0
                                       4                400                                         4.0
                                       5                500                                         5.0




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 47
                                                                          Policy # MI/SER/07/v01                           Page 9 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology




Figure 1: Reagent rack ―0‖ layout when running assays: HBsAG, HIV ½, HBcAb, HCV Ab & HTLV I/II




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 48
                                                                          Policy # MI/SER/07/v01                           Page 10 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology




Figure 2: Reagent rack ―1‖ layout when running assays: HBsAG, HIV ½, HBcAb, HCV Ab & HTLV I/II



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 49
                                                                          Policy # MI/SER/07/v01                           Page 1 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology




Figure 3: Reagent rack ―3‖ layout when running assays: HBsAG, HIV ½, HBcAb, HCV Ab & HTLV I/I I

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 50
                                                                          Policy # MI/SER/07/v01                           Page 12 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

                       v.      Once all the reagents and controls have been added to the racks, load the reagent
                               racks into the sample and reagent unit in the following order:

                                    1st ‗0‘- Large reagent rack
                                    2nd ‗1‘- Medium reagent rack
                                    3rd ‗3‘- Small reagent rack

                      vi.      If all five assays (HBsAG, HIV ½, HBcAb, HCV Ab & HTLV I/II) are going to
                               be run simultaneously, then the saved reagent template can be used. In the Load
                               dialog box, click the ‗Open Reagent Layout‘ button. Select the file name
                               ‗5assaytemplate.rea‘ and click ‗Open‘. All reagents and controls will be
                               automatically allocated to their assign position.

      22 Once all the required resources have been loaded onto the analyzer click ‗OK‘ and the Load
         dialog box will disappear. The analyzer will begin to check the resources to ensure their
         amounts are sufficient.

      23 The Load Plate dialog box will then appear for each assay respectively. Rename the plate
         ID with the corresponding assay name, for example HBsAg-, HIV-, HBC-,HCV- or HTLV-.
         The test date will automatically be added to the plate ID by the analyzer. When loading a
         BioRad assay microplate such as HBsAg and HIV ½ the following error message will appear
         when the plate is being renamed ―Plate ID is not valid for assay (….) Proceed Anyway?‖
         Click ‗Yes‘.

      24 Prepare the required number of microwell strips for the requested assay. The number of
         strips required is shown in the Plate Layout which is found on the right hand side of the Load
         Plate dialog box.

      25 Insert the microplate into the metal frame microplate holder. Make sure that the A1 position
         of the plate and holder match.

      26 Open the door to the microplate loading compartment and load the microplate and its holder
         onto the plate transport unit. Double check that the correct plate for the requested assay is
         being loaded. Once loaded, close the door to the microplate loading compartment.

      27 Once the plate is loaded click ‗OK‘. The Load Plate dialog box for the first assay will close
         and then reopen for the next required assay. Repeat steps 23 to 27 for each assay.

      28 After the last microplate is loaded, the analyzer will start to run automatically.



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 51
                                                                          Policy # MI/SER/07/v01                           Page 13 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

      29 From the Work folder click on ‗Schedule‘ to view the chorological order and completion time
         of the run. Take note of the amount of time required before operator intervention is needed
         (for OPD preparation and loading). Prepare the required OPD substrate in a 30ml Evolis vial
         before operation intervention is necessary and store the OPD in a dark location before loading
         is required.

      30 While the samples are being pipetted click on ‗Active Event Log‘ to ensure that all samples
         were pipetted successfully. Look for any red flags that may appear in the ―Dispense Sample‖
         section which indicates that an error has occurred. The analyzer will pause and flag with an
         error message if a clot or low sample volume is encountered, allowing for operator
         intervention.

      31 When operator intervention is required, the system will sound with an alarm and the
         message: ―Prepare [OPD for assay] and load in 1 minute‖. (The OPD substrate should
         already be prepared for the required assay before this message appears). Click ‗OK‘ to make
         this message disappear.

To Prepare Orthodiagnostic HBcore Ab, HCV Ab & HTLV I/II OPD Substrate:
HBcore Ab: 1 OPD tablet + 12 ml of Substrate Buffer for 1-7 strips
HCV Ab & HTLV I/II : 1 OPD tablet + 6 ml of Substrate Buffer for 1 – 3 strips
                     2 OPD tablet + 12 ml of Substrate Buffer for 4 – 7 strips
    32 After 1 minute the system will sound with another alarm and the Load dialog box will appear
       with the message: ‗Please load the requested items as soon as possible as the system is
       paused‘.

      33 Open the door to the sample and reagent unit and place the reagent without removing the
         reagent rack from the analyzer and load it into the correct location on the reagent rack as
         specified in the Load dialog box. Close the door to the sample reagent unit and click ‗OK‘.
         An alternative method would be to remove the reagent rack from the analyzer and load the
         correct OPD reagent into the position specified in the Load dialog box. Then re-insert the
         reagent rack onto the same track from where it was initially removed and click ‗OK‘. If this
         method is used a barcode error message may pop up when the rack is re-inserted click
         ‗Continue‘ and the run will not be affected.

      34 Repeat steps 31 to 33 for each time an operator intervention is required.

      35 The results report will automatically be printed after each assay is completed.

      36 Once all the assays are finished a dialog box prompting the removable of each plate and
         carrier from the analyzer will appear.


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 52
                                                                          Policy # MI/SER/07/v01                           Page 14 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

      37 Open the door to the microplate loading compartment and remove the microplate. After the
         microplate is unloaded close the door to the microplate loading compartment and click ‗OK‘.

      38 Repeat step 37 for each plate that needs to be unloaded from the analyzer.

      39 A blinking red LED light in the sample and reagent unit indicates that the reagent or sample
         racks can be unloaded from the analyzer.

      40 Remove all sample diluents from the dilution area.

      41 Do not unload pipette tip racks from the analyzer unless they are completely empty.

      42 Close the EVOLIS software. Select File | Exit from the menu bar or click the X icon at the
         top right-hand corner of the Evolis software and shut down the computer.

      43 Switch off the Evolis analyzer.

      44 Open the instrument cover and wipe the tip adapter (pipettor head) with 70% Ethanol wipes.

      45 Empty the liquid waste container.

      46 Empty the bag for the tip waste container and replace if damaged.

      47 Inspect the instruments (inner and outer surfaces) and racks for stains and spills. Clean if
         necessary.

      48 Weekly Maintenance Procedure :

                a. Run the weekly washer maintenance: (Procedure takes approximately 20 min)

                             i. Fill all wash buffer containers with de-ionized water
                            ii. Click ‗New Worklist‘ located in the upper toolbar
                          iii. The Set-up panel dialog box will appear
                           iv. In the Set-up Panel dialog box click ‗Add Plate‘
                            v. In the Set-up Panel dialog box click ‗Add Assay‘ and then select the file
                                 ‗WasherClean BR.asy‘, click ‗Open File‘.
                           vi. Click ‗OK‘ to validate the various dialog boxes until the worklist appears.
                          vii. Click ‗Start‘ (green button)
                         viii. Load an empty washer microplate with the metal plate holder onto the analyzer
                               when the Load Plate dialog box appears. Click ‗OK‘ after the plate is loaded
                               onto the analyzer.

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 53
                                                                          Policy # MI/SER/07/v01                           Page 15 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

                           ix. When the run is complete, unload the washer plate and replace the de-ionized
                               water with the appropriate wash buffers.

                    *The weekly washer maintenance does not have to be completed during the week
                     the monthly washer maintenance is being done.

                b. Decontaminate the pipettor wash station:

                            i. Pour 5 ml of decontamination solution consisting of 0.4% RIVASCOP (4ml of
                               RIVASCOP into 1L of water) into the pipettor wash station and let it soak for
                               a minimum of 15 minutes.
                           ii. Do not empty, the liquid will drain automatically when the system is initialized.

                  c. Decontaminate and wipe the tip ejection slide.

                 d. Clean the instrument surfaces and work area.


      49      Monthly maintenance Procedure:

                a. Run the monthly washer maintenance: (Procedure takes approximately 1 hour)

                          i.    Fill all the wash buffer containers with de-ionized water
                         ii.    Click ‗New Worklist‘ located in the upper toolbar
                        iii.    The Set-up panel dialog box will appear
                        iv.     In the Set-up Panel dialog box click ‗Add Plate‘
                         v.    In the Set-up Panel dialog box click ‗Add Assay‘ and then select the file
                               ‗WasherManifoldDisinfect BR.asy‘, click ‗Open File‘
                         vi.    Click ‗OK‘ to validate the various dialog boxes until the worklist appears.
                        vii.   Click ‗Start‘ (green button)
                       viii.   The Load dialog box will appear.
                         ix.   Pour 50ml of 0.4 % RIVASCOP into a 60ml of container and load the reagent
                               onto a ‗0‘ reagent rack. Then load the rack onto the analyzer.
                          x.   In the Load dialog box, allocate the bottle to the corresponding rack position in
                               which the container was loaded then click ‗OK‘.
                         xi.   Load an empty washer microplate with the metal holder onto the analyzer when
                               the Load Plate dialog box appears. Click ‗OK‘ after the plate is loaded onto the
                               analyzer.
                        xii.   A few minutes later a message saying to ‗open the drawer, unscrew waste bottle
                               1 and close the drawer‘ will appear.
                       xiii.   Unscrew the cap to waste bottle 1, which is located behind the wash buffer
                               bottles. Close the drawer when done and click ‗OK‘.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 54
                                                                          Policy # MI/SER/07/v01                           Page 16 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

                       xiv. After 15 minutes a message prompts you to re-open the drawer and re-screw the
                            cap to waste bottle 1, close the drawer when completed and click ‗OK‘.
                        xv. When the run is complete, unload the plate and the reagent rack and replace the
                            de-ionized water with the appropriate wash buffers.

                  b.    Decontaminate the sample and reagent racks, plate carrier and tip ejection slide.

                c.     Decontaminate the system liquid container

                       i. Empty the system liquid container.
                      ii. Empty the container and rinse thoroughly, twice with tap water and once with
                          de-ionized water.
                     iii. Inspect the filter (attached to the cap) to see if damaged or in need of replacing.
                     iv. Refill the container with freshly prepared System Liquid.
                          Preparation of System Liquid: 2 ml of Tween 20 to 10L of de-ionized water.
                d. Clean the washer buffer bottles only and NOT the caps & sensor devices.

                e.     Backup System Files
                          i. Click ‗Backup‘ button found at the upper tool bar.
                         ii. The System Backup dialog box will open up.
                        iii. Click ‗Backup System Files‘
                        iv. Once completed, click ‗Close‘ from the System Backup dialog box.

IV. Quality Control Procedures

        HBsAg
        1. Calculate the Mean Absorbance of the Negative Control(NCX):
           Sum of the absorbance values of the three negative controls divided by number of acceptable
           negative controls.
           Each individual absorbance must be >0.000 AU, and < or = to 0.150 AU. One Negative
           Control value may be discarded if it is outside this range. The NCX may be calculated from
           the two remaining absorbance values.
           Cutoff value= NCX+0.070
        2. Calculate the Mean Absorbance of the Positive Control(PCX):
           Sum of the absorbance values of the two Positive controls divided by two.
           The PCX must be > or = to 1.00 AU.
           Each individual absorbance must be within the range of 0.65 to 1.35 times the PCX.
           No Positive Control absorbance value may be discarded
        3. Calculate the Mean Absorbance of the Low Positive Control(LPCX):
           Sum of the absorbance values of the two Low Positive controls divided by two.
           The LPCX must be > or = to the Cutoff value. No Low Positive Control absorbance value
           may be discarded.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 55
                                                                          Policy # MI/SER/07/v01                           Page 17 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HbcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

HIV 1 & 2
     1. Calculate the Mean Absorbance of the Negative Control(NCX):
        Sum of the absorbance values of the three negative controls divided by number of acceptable
        negative controls.
        Each individual absorbance must be >0.000 AU, and < or = to 0.150 AU. One Negative
        Control value may be discarded if it is outside this range. The NCX may be calculated from
        the two remaining absorbance values.
        Cutoff value= NCX+0.250
     2. The absorbance value of the HIV-1 Positive Control must be > or = 0.700.

        3. The absorbance value of the HIV-2 Positive Control must be > or = 0.700.

        4. The absorbance value of the HIV-0 Positive Control must be > or = 0.700.

        If any failed control(s) that will invalidate the whole run, should inform Senior/Charge
        technologists and repeat testing.

Ortho HBcore Ab:
      1. Substrate Blank Acceptance: The Substrate Blank (A1 well) > = -0.020 and < = 0.05

          2. Negative Control Acceptance: Each Negative Control must < = 0.350 and > = -0.005.
             Negative Controls with absorbances between 0.000 and –0.005 are rounded to 0.000 for
             calculations. If one of the three Negative Controls is outside the acceptable range, the
             calculations are made based on the two acceptable Negative Controls. The plate is invalid
             and should be repeated, if two or more of the Negative Controls are unacceptable.
             Calculate the mean of the Negative Controls.
          3. Positive Control Acceptance: Both Positive Controls must be > = 0.800. Both Positive
             Controls must not differ by more than 0.500

          4. Cutoff Value Calculations: Cutoff = NC mean + 0.400

Ortho HCV Ab:
      1. Substrate Blank Acceptance: The Substrate Blank (A1 well) > = -0.020 and < = 0.05

          2. Negative Calibrator Acceptance: Each Negative Control must < = 0.120 and > = -0.005.
             Negative Calibrator with absorbances between 0.000 and –0.005 are rounded to 0.000 for
             calculations. If one of the three Negative Calibrators is outside the acceptable range, the
             calculations are made based on the two acceptable Negative Calibrators. The plate is
             invalid and should be repeated, if two or more of the Negative Calibrators are
             unacceptable.

          3. Calculate the mean of the Negative Calibrators.

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 56
                                                                          Policy # MI/SER/07/v01                           Page 18 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology

          4. Positive Control Acceptance: Both Positive Controls must be > = 0.800. Both Positive
             Controls must not differ by more than 0.600

          5. Cutoff Value Calculations: Cutoff = NCal mean + 0.600

Ortho HTLV-I/II Ab:

          1. Substrate Blank Acceptance: The Substrate Blank (A1 well) > = -0.020 and < = 0.05

          2. Negative Calibrator Acceptance: Each Negative Calibrator must < = 0.300 and > = -0.005.
             Negative Calibrators with absorbances between 0.000 and –0.005 are rounded to 0.000 for
             calculations. If one of the three Negative Calibrators is outside the acceptable range, the
             calculations are made based on the two acceptable Negative Calibrators. The plate is
             invalid and should be repeated, if two or more of the Negative Calibrators are
             unacceptable.
             Calculate the mean of the Negative Calibrators.
          3. Positive Control Acceptance: Both Positive Controls must be within range for plate to be
             valid.

          4. HTLV-I Positive Control > = 0.500
             HTLV-II Positive Control >= 0.500
          5. Cutoff Value Calculations: Cutoff = NCal mean + 0.150


V.     Reporting:

       Positive: >Cutoff Value
       Negative: < Cutoff Value
       All positive specimens will be spun and repeated x2 on the next run.
       Report as Positive if at least one of the repeats are > Cutoff Value.


VI. Troubleshooting Procedures

       For Flags, Recalculate Results and other procedures please refer to Evolis Short User Manual
       (Quick guide) Chapter 3: Advanced Information.
       For more information please refer to Evolis User Manual 1.90 EN (Long Version)




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 57
                                                                          Policy # MI/SER/07/v01                           Page 19 of 19
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: HBsAg, HIV 1/2, HBcAb, HCV Ab
                                                                          and HTLV I/II (cadaver/donor) Serology


VII. Reference

        Package insert from Bio-Rad HBsAg Assay
        Genetic Systems™ HBsAg EIA 3.0
        Revised June 2007

        Package insert from Bio-Rad Human Immunodeficiency Virus Types 1 and 2
        (Recombinant and Synthetic Peptides).
        Genetic Systems™ HIV-1/HIV-2 Plus O EIA
        Revised February 2008

        Bio-Rad Laboratories blood Virus division
        Redmond,WA 98052,U.S.A.
        U.S. License No.1109

       Package insert from Ortho®HCV 3.0 ELISA Test System, revised July 2003
       Package insert from Ortho® HBc ELISA Test System, revised June 2006
       Package insert from Ortho® HTLV-I/HTLV-II Ab-Capture ELISA Test System, revised Aug
       2003
        EVOLIS™ Short User Manual software version 1.90, revised June 2008
        EVOLIS™ User Manual software version 1.90, revised June 2007




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 58
                                                                          Policy # MI/SER/08/v04                          Page 1 of 9
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Epstein Barr Virus Serology
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: June 6, 2003
                                                                          Annual Review Date: January 10, 2011

                                             Epstein Barr Virus Viral Capsid Antigen IgG
I.        Introduction

          This test is used to detect the presence of EBV Viral Capsid Antigen (VCA) IgG antibodies in
          human serum using an ELISA system. In general, antibodies to EBV VCA appear within the
          first week after infection. The presence of EBV VCA IgG antibodies may indicate recent or
          past infection with EBV.

II.       Specimen Collection and Processing

       a. Approximately 10 ml of blood is collected in a red-topped tube.
       b. Centrifuge the tube at 2000 rpm for 10 minutes.
       c. Transfer serum into 2 ml storage tube and store in designated rack in 4 C fridge MIRT1.

III.      Procedure

          NEXGEN Analyzer
          i)      Reagents:
           Trinity Biotech Captia™ EBV VCA IgG
          1. Purified Recombinant VCA antigen ( a 47 kd fusion protein of 53 amino acids from the c-
              terminal half of p18)coated 96 wells microassay plate

          2. Serum Diluent Type 1:1 vial of 30 ml

          3. Cutoff Calibrator ( Calibrator)-labeled with CF(Correction Factor):1 vial of 0.4 ml

          4. High Positive Control: 1 vial of 0.8 ml

          5. Low Positive Control: 1 vial of 0.8 ml

          6. Negative Control: 1 vial of 0.8 ml

          7. Horseradish-peroxidase Conjugate: 1 vial of 16 ml, ready for use

          8. Chromogen/Substrate Solution Type 1(Tetranethylbenzidine): 1 vial of 15ml, ready for use.
             The reagent should remain closed when not in use. If allowed to evaporate, a precipitate
             may form in the reagent well.



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 59
                                                                          Policy # MI/SER/08/v04                          Page 2 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Epstein Barr Virus Serology

          9. Wash Buffer Type 1(X20): 1 bottle of 60 ml, dilute 1 part concentrate to 19 parts of D
              H2O.
          10. Stop Solution ( H2SO4): 1 bottle of 15 ml, ready for use.

          ii)       Method:
                    NEXGEN Four™ Analyzer

          i)        Reagent Preparation:
                   1. Prepare wash buffer: 60 ml of 20X Wash Buffer
                       Concentrate + 1140 ml of D H2O. Store at 4 0C. Fill up EBV
                       working buffer bottle in NEXGEN Analyzer when needed.

                   2.     Do not need to transfer Cutoff Calibrator(Blue), High Positive Control(Red) ,Low
                          Positive Control(Clear) and Negative Control (Green) to curvettes.

                   3.     Pour Diluent into NEXGEN large -sized container and labeled as
                           'Diluent'. Fill up almost to the top.
                          Pour Conjugate into NEXGEN medium -sized container and labeled as
                           'Conjugate'.
                          Pour Substrate into NEXGEN medium-sized container and labeled as
                          'Substrate'. Keep substrate in the dark until ready to load in NEXGEN.
                          Pour Stop Solution into NEXGEN medium-sized container and labeled as
                          'Stop Solution'.
                          All these reagents are stored in the respective containers in 40C
                           refrigerator.
                           Use new containers for each new Lot #. Change container if needed.

                   4.      Bring microwell pouch and the above mentioned containers to room temperature
                          before use.

                   5.     Empty waste every day. Change DH2O and rinse container monthly.

        ii)   Assay Method:
     1. Turn NEXGEN and the computer‖ ON‖ at the beginning of the day. Sign in at the Support
        prompt. Type in ―qw‖ and Enter.

     2. Allow instrument and PC to communicate for about 15 minutes.

     3. Replace the water Tank 1 (Buffer 1) on the left side with the EBV Wash Buffer bottle.



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 60
                                                                          Policy # MI/SER/08/v04                          Page 3 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Epstein Barr Virus Serology

     4. Click on or touch icon labeled ―Maintenance” on the left vertical panel.

     5. ―Performing Self Test‖ will appear on screen. Once Self test is done,‖ Self Test‖ square will
        be highlighted. Click on or touch down arrow in Resource field to see that all tests have passed.

     6. Click on or touch ―Print Report‖. Staple self test printout with worklist and result sheet.

     7. Click on or touch‖ ―Hydraulic filling ―-left side three times, wait for the procedure to finish.
        Check for the presence of bubbles in the lines,

     8. Click on or touch ―Washer priming‖- left side once.

     9. Click on or touch ―Manual Controls‖ tab. To fill tips, push on the tips rack. Fill the tips
        container. (Do not use the 4 rows of tips close to the front panel of the machine. Instrument
        will give ―Tip Crash‖ error.) Click or touch ―Left‖ under ―Zero Tip‖. This will reset tip
        counter.

     10. To move the wanted sector, click on or touch the Number 1 in the carousel. The #1 sector then
         will be moved to the front. Continue with #2 and #3 if needed. Remove enough sectors for
         testing.

     11. Click on or touch ―Close‖ at bottom right of screen.

     12. Go to ―Session Builder—Create or modify ELISA session for NexGen‖ and click on or touch
         screen. Select panel from the ―Select panel or Session‖ field-click on or touch
         Trinity_EBV_PlateA. Click on or touch ―Next” on the bottom right of screen.

     13. ―Information‖ screen appears, can update Kit lot # by click on or touch‖ New Kit lot‖. Enter
         new lot #, expiration date and CF (Correction factor on Calibrator vial).Click on or
         touch―Update and Apply‖. Click on or touch ―Next‖.

     14. Click on space beside Sample Code. Scan the specimen barcode in, remove lid, and load the
         specimen in proper position in #1 sector. Repeat until all specimens have been scanned and
     loaded into sectors. Make sure that the tubes are in the upright position, not leaning against
     the side of the holder. Load corresponding dilution tubes at the back of the each segment.

     15. Click on or touch Test Name to select tests for all the samples, orange dots will appear under
         test. Click on or touch ―Next‖.

     16. Click on or touch ―Add All‖. Click on or touch ―Next x 2.



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 61
                                                                          Policy # MI/SER/08/v04                          Page 4 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Epstein Barr Virus Serology

     17. Highlight ―I want to build the created session and run immediately”. Click on or touch
         ―Finish”(NOT ‘CLOSE’). ―Initializing instrument‖ appears on screen.‖ Interlock sensor
         disable‖, Click on or touch ―OK‖. Procedure Form‖ appears, Click on or touch ―OK‖ x 2
         re: message ―No error‖.

     18. To load sectors: (open blue sliding door in front)
         Click on or touch ―Right/Left‖ to move Sector #1to the front. Load Sector #1. Click on or
         touch‖ Right‖ to move to sector #. Repeat until all sectors are loaded. Close door.

     19. Click on or touch‖OK‖ x 2.

     20. Click or touch ―Down Arrow‖:
         Highlight :Microplate A-will show # of wells/strips needed for this run, fill in blanks if
         required.
         Highlight : Reagent Holder left-will show position/amount needed for the run for each
         controls, cutoff calibrator, conjugate, substrate, stop solution, diluent, and dilution curvettes.
         1st strip- blank, NC, Cutoff calibrator x3,HP ,LP and specimen.
     21. Load reagents, controls and strips(plate):
         Push on the reagent door to open the reagent area.
         Loading Reagents: 1 Negative Control                 2 Dilution Curvette
                             4 Cutoff Calibrator             5 Dilution Curvette
                             7 High Positive Control        8 Dilution Curvette
                             10 Low Positive control        11 Dilution Curvette
                             R1 Conjugate
                             R2 Substrate
                             R3 Stop Solution
                             R5 Diluent
         Load substrate last, after the plate goes in for 1st incubation.

     22. Load plate:
         Remove plastic plate holder from ‗A‘. Slide plate in with strips in the front. Make sure plate is
         secured by the metal plate holder. First strip always contains- Blank, Negative Control,
         Calibrator x2, High Positive Control and Low Positive Control and 1 spot for specimen. Fill in
         blank wells if needed.

     23. When everything is loaded, click on or touch ―Next‖ at the bottom right of the screen.

     24. Click on or touch ―Run”.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 62
                                                                          Policy # MI/SER/08/v04                          Page 5 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Epstein Barr Virus Serology


     25. When the test is done, there is a ―Circled” OK in the middle of the screen. Click on or touch it.

     26. Click on or touch ―Next‖.

     27. Make sure ―I want to validate and archive the executed session‖ is selected. Click on or touch
         ―Finish‖ at bottom right of screen. ―Closing instrument‖ appears on screen.

     28. Click on or touch ―ELISA Data Reduction‖ at bottom left of screen.

     29. Find session in ―Recent available sessions‖ at top of page.

     30. Click on or touch session to highlight your session. Click or touch ―Next‖ at bottom right of
         screen.

     31. Click on or touch ―Report‖ tab at top of screen.

     32. Click onor touch highlighted Trinity_EBV_Plate A.

     33. If test passed, ―VALID TEST‖ will scroll across top of page.

     34. Click on or touch ―Print‖ at top of page. After the report is printed, click or touch ―Close‖.

     35. If need to run another assay, click on or touch ―Maintenance‖:
                  a. Change buffer and click or touch‖ Manual Control‖.
                  b. Click on or touch ―Left Tank 1‖under―Washer Priming ―once.
                  c. Click on or touch on sector # to remove specimens from the previous run.
                  d. Click on or touch ―Close‖.
                  e. Click on or touch‖ Session Builder‖ to start another session.

     36. Upon completion of the run, replace the wash tank solutions with distilled water and flush the
         system by the shutdown procedure.

     37. Click on or touch Monitor on the left column.

     38. Perform Daily Maintenance: click on or touch ―NEXT‖ in ―Clean Up‖ on the upper right of the
         screen. NOT the “Next” on the bottom right of the screen.

     39. Empty the used tips drawer, disinfect with Virovox and wipe dry. Click on or touch‖ Next‖.

     40. Empty the waste tank by pressing the waste button on the left-hand side of the instrument.
         Press the button until no more waste draining out. Click on or touch‖ Next‖.

     41. ―Replace all Buffers with DH2O‖. Click on or touch‖ Next‖.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 63
                                                                          Policy # MI/SER/08/v04                          Page 6 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Epstein Barr Virus Serology

     42. Remove sample tubes using the manual control tab. Click on Sector # 1, open the door and
         remove Sector #1, continue until all sectors in use are removed. Discard sample tubes in yellow
         discard container. Return sector to its position in reverse order.

     43. ―Remove all reagents from the reagent tray‖. Wash the tray in case of spills. Disinfect if
         material potentially contaminated. Click on or touch‖ Next‖.

     44. ―Remove all Microplates‖. Click on or touch‖ Next‖.

     45. Click or touch ―Parking Position‖.

     46. Remove all sample tubes from the sample rotating carousel. Wash the carousel with
         disinfectant in case of potentially contaminated spills.

     47. Click on or touch: Both- ―Washers Cleaning‖ for 5 times.

     48. ―Shut Down‖ button will be highlighted. Click on or touch it. Wait for‖ Shut Down Complete‖
         to appear on the screen. It is now safe to switch the instrument off‘, click or touch ―OK‖. Turn
         off NEXGEN.

     49. Then click on or touch ―Close‖x2, then click or touch ―Exit‖.

     50. Click on ―Start‖ on lower left corner, click on ―Turn off computer‖, and click on ―Turn Off‖.

     51. If for some reason the NexGen was not shut down properly ( screen frozen or no
        communication between NexGen and computer..etc):

                     a. Click or touch ―EXIT‖ if allowed to do so.
                     b. Double click on:‖VNC server‖ Icon, will show: Nexgen_076, click ―OK‖.
                     c. Enter Session password -―admin‖.
                     d. Then the screens will show lot of windows, close all windows by clicking on each
                        of the ―x‖ at the right top corner.
                     e. Click or touch ―Start‖ at the lower left corner. Click on‖ Turn off computer‖ and
                        click on ―Turn Off‖.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 64
                                                                          Policy # MI/SER/08/v04                          Page 7 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Epstein Barr Virus Serology

      52. Weekly Maintenance Procedure :
          Click or touch‖ Maintenance‖, then the ―Weekly Maintenance‖ Tab and follow the steps by
          clicking on ―Next ―for each step:
                         a. Empty, wash and refill all tanks with fresh DH2O.
                         b. Flush the tanks with ―End of Work‖ function.
                         c. Empty and wipe the used tips tray with disinfectant.
                         d. Wipe the external surface of the needle with 70% alcohol.
                         e. Remove all reagents from the reagent rack.
                         f. Clean the reagent rack with disinfectant.
           Initial in NEXGEN Maintenance Book.

      53. Monthly maintenance Procedure:
          Click or touch‖ Maintenance‖, then the ―Monthly Maintenance‖ Tab and follow the
          Steps by clicking on ―Next‖ for each step.
                  f. Pour 50 mL of AXSYM® Tubing Decon Solution (7805-09) into a clean beaker
                      containing 950 ml of DH2O. Fill Tank 2(both sides) with 500 ml of working
                      solution each.
                  g. Circulate 300 ml of liquid from Tank 2 (both sides).
                  h. Replace the contents of Tank 2 with DH2O (both sides).
                  i. Circulate 600 ml of liquid from Tanks 2 (both sides).
                  j. Empty the waste tank by pressing the waste button of the instrument.
                  k. Empty waste.
           Initial in NEXGEN Maintenance Book.

IV.       Assay validation

          Absorbencies (A) are read at 450nm

           Cutoff calibrators and controls must be run with each run.

           Reagent Blank must be <0.150 A

           Negative Control must be < 0.250 A

           Each Cutoff calibrator must be > or =0.250A

           High Positive Control must be > or =0.500




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 65
                                                                          Policy # MI/SER/08/v04                          Page 8 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Epstein Barr Virus Serology

V.        Calculations

          Mean Cutoff Calibrator A: ∑ Cutoff Calibrator x3
                                              3

           If any of the Cutoff Calibrator values differ by more than 15% from the mean, discard
           that value and calculate the average of the two remaining values.

           Correction Factor is printed on the Cutoff Calibrator vial and it is different for each lot.
           Cutoff Calibrator value-is determined by multiplying the correction factor by the Mean
           Cutoff Calibrator value.

           ISR (Immune Status Ratio) :               Specimen OD value
                                                    Cutoff Calibrator value


VI.       Interpretation of Results

          Spectrophotometer Single Wavelength (450 nm)

          Negative:            ISR ≤ 0.90

          Equivocal:           ISR 0.91-1.09

         Positive:             ISR > 1.10


VII.      Reporting:

          Positive by EIA

          Negative by EIA

          Equivocal by EIA




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 66
                                                                          Policy # MI/SER/08/v04                          Page 9 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Epstein Barr Virus Serology


VIII. Quality Control:

          Cutoff calibrator is used to calibrate the assay to account for day-to-day fluctuations in
          temperature and other testing condition

           Run external control (Pooled positive specimens-also sent to PHL for confirmatory
           testing) with each new lot.

           CAP provides external proficiency testing.


IX.       References:

                    1. Package Insert of Captia ™ EBV VCA IgG ( Trinity BioTech) P/N 4700-29 Rev K,
                       issued October 2004.
                    2. NEXGEN FOUR ™ User‘s Manual, version1.3-Rev.D
                    3. NEXGEN FOUR ™ software version 2.1 SP2.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 67
                                                                          Policy # MI/SER/14/v02                          Page 1 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay
Issued by: LABORATORY MANAGER                                             Original Date: November 18, 2009
Approved by: Laboratory Director                                          Revision Date:
                                                                          Annual Review Date: January 10, 2011

                                        Aspergillus Galactomannan Antigen Detection Assay

I.        Introduction

          The PlateliaTM Aspergillus EIA is a qualitative enzyme immunoassay for the detection of
          Aspergillus galactomannan antigen in serum and Bronchoalveolar Lavage (BAL) samples.


II.       Specimen Collection and Processing

          The test is performed on serum and BAL samples. Blood is collected in a serum separator tube
          and separated by centrifugation at 3000 rpm for 10 minutes. After initial opening, samples
          may be stored at 2-8oC for up to 48 hours prior to testing. For longer storage, store the serum
          at -70oC.


III.      Procedure

               EVOLIS ™ Analyzer

          i)Reagents:

               BioRad Aspergillus Galactomannan kit:
               1. Galactomannan monoclonal antibodies coated microwell plate
               2. Concentrated wash solution (10X)
               3. Negative control serum (Human)
               4. Cut-off control serum (Human)
               5. Positive control serum (Human)
               6. Conjugate – Anti-galactomannan monoclonal antibody/ peroxidase labeled
               7. Serum treatment solution (EDTA acid solution)
               8. TMB substrate buffer
               9. Chromogen TMB solution
               10. Stopping solution – 1.5N sulphuric acid


        ii)Method:
             The Platelia Aspergillus EIA for the detection of Aspergillus galactomannan antigen in
             serum and BAL samples can be run on the EVOLIS ™ analyzer.

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 68
                                                                          Policy # MI/SER/14/v02                          Page 2 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay

Negative, Cut-Off and Positive Control Preparation

The Negative, Cut-Off and Positive controls are provided as freeze-dried samples in the
Galactomannan assay kit and need to be reconstituted before their use:
           1. The control serums should be rehydrated just prior to testing.

                2. Reconstitute each bottle of control with 1mL of purified water.

                3. Mix thoroughly after allowing 2 to 3 minutes for rehydration of the control.

                4. Aliquot 390ul of each control into a 2ml sample tube and label the tube appropriately.

                5. Freeze any remaining control that will not be used after rehydration at -20oC.

                6. Controls that have been previously rehydrated and frozen at -20oC may be thawed and
                   do not require further rehydration. Frozen control samples may be stored at -20oC for
                   up to 5 weeks.

                    *Follow steps 1 to 6 for each bottle of control that needs to be reconstituted.

Pre-treatment of the serum
Serum and BAL samples need to be heat-treated in the presence of EDTA prior to being tested on the
analyzer. This pre-analytical procedure helps dissociate immune complexes and causes serum proteins
to precipitate which can possibly interfere with the testing procedure.
All control serum: negative, cut-off and positive must be processed at the same time and in the same
manner as the patient samples:
             1. Label 2 empty 2ml sample tubes with the patient barcode label. For the control
                samples label 2 empty tubes for each: Neg, Cut-Off & Pos, respectively.

                   *The Cut-Off Control will be sampled twice by analyzer but only one sample tube
                   needs to be prepared.
                2. Pipette 390ul of each test serum/BAL or control into one of the 2 ml sample tubes.

                3. Add 130ul of serum treatment solution into the sample tube.

                4. Tightly close the caps of the tubes to prevent opening during heating.

                5. Mix tubes thoroughly by vortexing each sample.

                6. Heat the sample tubes by placing them in a heating block set at 120oC for 6 minutes.
                   The tubes should only be placed in the heating block only after 120oC has been
                   reached.


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 69
                                                                          Policy # MI/SER/14/v02                          Page 3 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay


                7. Carefully remove the heated tubes from the heating block after 6 minutes and place
                   them in the centrifuge. Centrifuge tubes at 10,000 x g for 10 minutes.

                8. Transfer 300ul of the treated serum or BAL supernatant into the second labeled 2ml
                   polypropylene sample tube which will be placed on to the EvolisTM analyzer.

                9. After preparation the supernatant may be removed and stored at 2-8 oC for up to 48
                   hours prior to testing. If analysis of the results indicates that retesting of a sample is
                   required, another aliquot of the serum or BAL must be heat treated for retesting.

Before beginning EIA assay:
           j. Fill up wash buffer containers:

                    Galactomannan BioRad Wash
                    a) Prepare when necessary:
                       Add 100 ml of 10x BioRad wash solution to 900 ml of distilled water.
                    b) Fill up Evolis wash container with red tubing.

                   De-ionized Water:
                   c) Fill up Evolis container with blue tubing with de-ionized water.
                b. Working TMB substrates are prepared fresh during each run in a 30ml EVOLIS vial.

                c. The Aspergillus Galactomannan conjugate and stopping solution will be pipette from
                   their original containers and can loaded onto the analyzer as is.

                d. All controls: negative, cut-off and positive serums need to be rehydrated and heat
                   treated prior to their use as outlined in the pre-treatment procedure and the supernatant
                   is transferred into a labeled 2ml polypropylene sample tube.

                e. Bring all required microwell strips and the reagents mentioned above to room
                   temperature before use.

                f. Empty the waste tank and check the system liquid container (refill if necessary).
                   System Liquid Preparation: 2ml of Tween 20 to 10L of de-ionized water.


     1. Turn ―ON‖ the EVOLIS analyzer first and then the computer at the beginning of the day.

     2. Double click on the EVOLIS icon located on the computer desk-top.

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 70
                                                                          Policy # MI/SER/14/v02                          Page 4 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay


     3. Log onto the system by clicking on ―OK‖, no password is required.

     4. A self test of the system is automatically initialized each time the EVOLIS software is run. The
        self test is considered satisfactory if the word ―PASSED‖ appears beside each instrument
        module. Print a copy of the self test report and combine it with the worklist and result sheets.

     5. Prepare specimens by removing the lids off the pre treated serum supernatant and load all
        sample tubes with the barcode facing right on to the sample rack (rack code T). Check the
        quality of the samples by ensuring all clots, foam and bubbles have been removed.

     6. Open the door to the sample and reagent unit. Load the sample rack with the bar-codes facing
        the bar-code reader onto the track marked by the solid red LED light.

     7. The ‗Patient Editor‘ dialog box will appear with the specimen numbers from the loaded rack.
        Double check that the specimen numbers are correct. If there is a blank space under the patient
        ID column, click on the space and enter the specimen number manually. All specimen
        numbers entered manually will be flagged with the code ―ManID‖ on the results report.

     8. Choose the Galactomannan assay from the drop down menu which appears along the top of the
        Patient Editor dialog box:

                    Galactomannan assay = “MT SINAI Aspergillus EIA New BR V1‖

     9. Click on the assay box corresponding to the sample number if you want that assay to be run for
        that sample. A check mark is shown for a sample where the assay has been selected.

     10. Once the required assay has been selected for each sample click ‗Close‘ to save. A solid LED
         light will show up for the next available track in the sample and reagent unit which indicates
         where the next sample rack can be loaded. Repeat steps 6 to 11 for each new sample rack that
         is loaded on to the analyzer.

     11. A total of four sample racks need to be loaded on to the analyzer even if all the sample racks
         are not required in order for the reagent template to be used in a later step. When an empty
         sample rack is loaded on to the analyzer just click ‗Close‘ when the Patient Editor dialog box
         appears.

     12. To check and validate the worklist click on the ‗New Worklist‘ icon located on the upper
         toolbar and the Set-Up dialog box will appear.



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 71
                                                                          Policy # MI/SER/14/v02                          Page 5 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay

     13. Click on ―+‖ beside the assay file name that is associated with the plate. The plate layout will
         appear at the right hand side of the dialog box and will show the total number of controls and
         specimens to be tested, their assigned wells and the number of strips required for the assay.

     14. If duplicate samples need to be tested for a given assay in the same run:
                   vi.  From the Patient Editor dialog box select the assay folder in which the
                        duplicate sample needs to be run and click ‗Add Patient‘.
                  vii.  The Select Patient(s) dialog box will appear.
                 viii. Check the Allow multiple determinations box which appears at the bottom of
                        the Select Patient|(s) dialog box.
                   ix. The patient sample IDs will appear. Click on the sample ID in which multiple
                        testing is required and click OK.
                    x. In the Set-up Panel dialog box, a (x2) will appear beside the patient ID under
                        the assay folder in which it was selected.

     15. If all the information is correct click ‗OK‘ to validate the worklist.

     16. A separate dialog box asking for the reagent lot number will appear for each assay that is
         ordered. Double check the lot number and expiry date and click ‗OK‘ if everything is correct.

     17. Click on the ―+‖ to expand the Work folder and click on ‗Plate Layout‘, the number of
         microwell strips required for the assay can be reviewed. Click the ‗Print‘ button located along
         the top of the toolbar to print the plate layout if desired.

     18. Click ‗START‘ (green button) located on the upper tool bar to open the Load dialog box.

     19. The Load dialog box appears and shows where to load all the required resources.

     20. Load all required resources from left to right:

                          i. Pipette tips: Grey = 1100 ul tips (full rack)
                                           Brown= 300 ul tips (full rack)
                                           Red = Incomplete tip rack

                            *The pipette tips have to be loaded in the exact position as shown in the
                            layout
                         ii. Load the assay reagents on the reagent racks according to the following
                              templates (Figure 1-2). The reagent template specific for the Aspergillus
                              Galactomannan assay should be used.

                        iii. Ensure all caps on the reagent/control containers have been removed before
                             loading them onto the analyzer.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 72
                                                                          Policy # MI/SER/14/v02                          Page 6 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay

TMB Substrate preparation for Aspergillus Galactomannan assay (prepare in 30ml coated vial):
     Prepare Fresh (Same Day) Working Chromogen as follows (always make 1 extra strip):

                                   Number of Strips Chromogen Reagent (uL) Chromogen Diluent (mL)
                                       1                   40                     2.0
                                       2                   80                     4.0
                                       3                  120                     6.0
                                       4                  160                     8.0
                                       5                  200                    10.0




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 73
                                                                          Policy # MI/SER/14/v02                          Page 7 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay




Figure 1: Reagent rack ―-1‖ layout when running assays: Galactomannan assay


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 74
                                                                          Policy # MI/SER/14/v02                          Page 8 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay




Figure 2: Reagent rack ―3‖ layout when running assays: Galactomannan Assay
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 75
                                                                          Policy # MI/SER/14/v02                          Page 9 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay

                         iv. Once all the reagents and controls have been added to the racks, load the reagent
                             racks onto the sample and reagent unit in the following order:

                                    1st =‗1‘- Medium reagent rack
                                    2nd=‗3‘- Small reagent rack

                          v. In the Load dialog box, click the ‗Open Reagent Layout‘ button. Select the file
                             name ‗galactomannan.rea‘ and click ‗Open‘. All reagents and controls will
                             be automatically allocated to their assign position.

     21. Once all the required resources have been loaded onto the analyzer click ‗OK‘ and the Load
         dialog box will disappear. The analyzer will begin to check the resources to ensure their
         amounts are sufficient.

     22. The Load Plate dialog box will then appear. Rename the plate ID with the corresponding
         assay name, for example Galactomannan. The test date will automatically be added to the
         plate ID by the analyzer.

     23. Prepare the required number of microwell strips for the requested assay. The number of strips
         required is shown in the Plate Layout which is found on the right hand side of the Load Plate
         dialog box.

     24. Insert the microplate into the metal frame microplate holder. Make sure that the A1 position of
         the plate and holder match.

     25. Open the door to the microplate loading compartment and load the microplate and its holder
         onto the plate transport unit. Once loaded, close the door to the microplate loading
         compartment.

     26. Once the plate is loaded click ‗OK‘. After the microplate is loaded, the analyzer will start to
         run automatically.

     27. From the Work folder click on ‗Schedule‘ to view the chorological order and completion time
         of the run.

     28. While the samples are being pipetted click on ‗Active Event Log‘ to ensure that all samples
         were pipetted successfully. Look for any red flags that may appear in the ―Dispense Sample‖
         section which indicates that an error has occurred. The analyzer will pause and flag with an
         error message if a clot or low sample volume is encountered, allowing for operator
         intervention.

     29. The results report will automatically print out after the assay is completed.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 76
                                                                          Policy # MI/SER/14/v02                          Page 10 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay

     30. Once the assay is finished a dialog box prompting the removable of the test plate and carrier
         from the analyzer will appear.

     31. Open the door to the microplate loading compartment and remove the microplate. After the
         microplate is unloaded close the door to the microplate loading compartment and click ‗OK‘.

     32. A blinking red LED light in the sample and reagent unit indicates that the reagent or sample
         racks can be unloaded from the analyzer.

     33. Do not unload pipette tip racks from the analyzer unless they are completely empty.

     34. Close the EVOLIS software. Select File | Exit from the menu bar or click the X icon at the top
         right-hand corner of the Evolis software and shut down the computer.

     35. Switch off the Evolis analyzer.

     36. Open the instrument cover and wipe the tip adapter (pipettor head) with 70% Ethanol wipes.

     37. Empty the liquid waste container.

     38. Empty the bag for the tip waste container and replace if damaged.

     39. Inspect the instruments (inner and outer surfaces) and racks for stains and spills. Clean if
         necessary.

     40. Weekly Maintenance Procedure :

             a. Run the weekly washer maintenance: (Procedure takes approximately 20 min)

                      Fill all wash buffer containers with de-ionized water
                      Click ‗New Worklist‘ located in the upper toolbar
                      The Set-up panel dialog box will appear
                      In the Set-up Panel dialog box click ‗Add Plate‘
                      In the Set-up Panel dialog box click ‗Add Assay‘ and then select the file
                       ‗WasherClean BR.asy‘, click ‗Open File‘.
                      Click ‗OK‘ to validate the various dialog boxes until the worklist appears.
                      Click ‗Start‘ (green button)
                      Load an empty washer microplate with the metal plate holder onto the analyzer when
                       the Load Plate dialog box appears. Click ‗OK‘ after the plate is loaded onto the
                       analyzer.
                      When the run is complete, unload the washer plate and replace the de-ionized water
                       with the appropriate wash buffers.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 77
                                                                          Policy # MI/SER/14/v02                          Page 11 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay

                      *The weekly washer maintenance does not have to be completed during the week
                       the monthly washer maintenance is being done.

             b. Decontaminate the pipettor wash station:

                      Pour 5 ml of decontamination solution consisting of 0.4% RIVASCOP (4ml of
                       RIVASCOP into 1L of water) into the pipettor wash station and let it soak for a
                       minimum of 15 minutes.
                      Do not empty, the liquid will drain automatically when the system is initialized.

             c. Decontaminate and wipe the tip ejection slide.
             d. Clean the instrument surfaces and work area.

     41. Monthly maintenance Procedure:

          a. Run the monthly washer maintenance: (Procedure takes approximately 1 hour)

                      Fill all the wash buffer containers with de-ionized water
                      Click ‗New Worklist‘ located in the upper toolbar
                      The Set-up panel dialog box will appear
                      In the Set-up Panel dialog box click ‗Add Plate‘
                      In the Set-up Panel dialog box click ‗Add Assay‘ and then select the file
                       ‗WasherManifoldDisinfect BR.asy‘, click ‗Open File‘
                      Click ‗OK‘ to validate the various dialog boxes until the worklist appears.
                      Click ‗Start‘ (green button)
                      The Load dialog box will appear.
                      Pour 50ml of 0.4 % RIVASCOP into a 60ml of container and load the reagent onto a
                       ‗0‘ reagent rack. Then load the rack onto the analyzer.
                      In the Load dialog box, allocate the bottle to the corresponding rack position in
                       which the container was loaded then click ‗OK‘.
                      Load an empty washer microplate with the metal holder onto the analyzer when the
                       Load Plate dialog box appears. Click ‗OK‘ after the plate is loaded onto the
                       analyzer.
                      A few minutes later a message saying to ‗open the drawer, unscrew waste bottle 1
                       and close the drawer‘ will appear.
                      Unscrew the cap to waste bottle 1, which is located behind the wash buffer bottles.
                       Close the drawer when done and click ‗OK‘.
                      After 15 minutes a message prompts you to re-open the drawer and re-screw the cap
                       to waste bottle 1, close the drawer when completed and click ‗OK‘.
                      When the run is complete, unload the plate and the reagent rack and replace the de-
                       ionized water with the appropriate wash buffers.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 78
                                                                          Policy # MI/SER/14/v02                          Page 12 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay

       b. Decontaminate the sample and reagent racks, plate carrier and tip ejection slide.

        c. Decontaminate the system liquid container
                Empty the system liquid container.
                Empty the container and rinse thoroughly, twice with tap water and once with de-
                 ionized water.
                Inspect the filter (attached to the cap) to see if damaged or in need of replacing.
                Refill the container with freshly prepared System Liquid.
                Preparation of System Liquid: 2 ml of Tween 20 to 10L of de-ionized water.

       d. Clean the washer buffer bottles only and NOT the caps & sensor devices.

       e. Backup System Files
            Click ‗Backup‘ button found at the upper tool bar.
            The System Backup dialog box will open up.
            Click ‗Backup System Files‘
            Once completed, click ‗Close‘ from the System Backup dialog box.


IV. Quality Control Procedures

     1. Calculate the mean absorbance of the Cut-Off Control (Cut-Off X): Sum of the O.D values for
        the two Cut-Off controls divided by 2.

          The O.D of each Cut-Off control must be ≥0.3000 and ≤0.8000 to be considered valid.

     2. Calculate the Negative Control Index by dividing the O.D of the Negative Control by the mean
        absorbance of the Cut-Off Control.

                    Negative Control Index = O.D Negative Control
                                            Mean Cut-off Control O.D

          The index of the Negative Control Serum must be less than 0.40 to be considered valid.

     3. Calculate the Positive Control Index by dividing the O.D of the Positive Control by the mean
        absorbance of the Cut-Off Control.

                    Positive Control Index= O.D Positive Control
                                          Mean Cut-off Control O.D

           The index of the Positive Control Serum must be greater than 2.00 to be considered valid.

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 79
                                                                          Policy # MI/SER/14/v02                          Page 13 of 13
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Aspergillus Galactomannan
                                                                                         Antigen Detection Assay


V. Reporting:

     1. The presence or absence of galactomannan antigen in the test sample is determined by the
        calculation of an index for each patient sample. To calculate the Index of a sample, divide the
        O.D of the patient sample by the mean O.D of the cut-off control.

                    Patient Serum/BAL Index = O.D Patient Sample
                                           Mean Cut-off Control O.D

     2. Patient serums/BAL with an index < 0.50 is considered to be negative for galactomannan
        antigen.

     3. Patient serums/BAL with an index ≥ 0.50 is considered to be positive for galactomannan
        antigen.

     4. All positive specimens will be spun and repeated x2 on the next run.

     5. An absorbance value of less than 0.000 may indicate a procedure or instrument error and the
        result is considered invalid and the specimen should be re-run.


VI. Reference:

        Package insert from Platelia Aspergillus EIA
        Revised July 2007
        EVOLIS™ Short User Manual software version 1.90, revised June 2008
        EVOLIS™ User Manual software version 1.90, revised June 2000




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 80
                                                                          Policy # MI/SER/09/ v04                         Page 1 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Human T-Lymphotropic Virus
                                                                                         Type 1 (HTLV-1) EIA for
                                                                                         TGLN Call Back Only
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: July 23, 2006
                                                                          Annual Review Date: January 10, 2011

           Human T-Lymphotropic Virus Type 1 (HTLV-1) EIA for TGLN Call Back Only

I.        Introduction

          The Adaltis DETECT-HTLV EIA Test Kit is a solid phase enzyme immunoassay utilizing a
          mixture of synthetic peptides for the detection of antibodies to HTLV-I and II in human serum
          or plasma.

II.       Specimen Collection and Processing

          Blood is collected (5 mL for adults and l mL for neonates) in a serum separator tube and
          separated by centrifugation. The serum is stored at 4oC.

III.      Procedure

          i)        Reagents:

                    Positive Control
                    Negative control
                    Anti-Human IgG Peroxidase Conjugate
                    Conjugate Diluent
                    Tetramethylbenzidine (TMB) Reagent
                    Peroxide Reagent
                    Stop Solution
                    2 x 96 well Microplate coated with HTLV-I & HTLVII peptide


          ii)       Other Materials :

                    P Lab Analyser System
                    5 mL graduated Pipettes
                    10 mL graduated Pipettes
                    1 mL graduated Pipettes
                    10-100 uL pipettor
                    Pipette tips
                    55 x 12 (3.5 mL) tubes
                    Automatic pipettor

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 81
                                                                          Policy # MI/SER/09/ v04                         Page 2 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Human T-Lymphotropic Virus
                                                                                         Type 1 (HTLV-1) EIA for
                                                                                         TGLN Call Back Only

          iii)      Method:

A. Using P-Lab (Personal Lab)

         Before beginning EIA assay:
     1. Allow all components to reach room temperature.
        Prepare wash solution:
        Add 100 ml of 25x wash solution to 2400 ml of distilled water.
        Fill up P-Lab wash bottle. Place bottle in ‗Buffer #1‘ position in P-Lab.
        Turn hard drive and monitor. Wait for software to be fully loaded before turning the P-Lab
        ‗ON‘.

     2. Click once on third square from the left: ‗open session‘.. Click on ‗New‖. Type in assay &
        date performed, e.g. HTLV 280499(HTLV tested April 28,99). Note session # e.g.‘31‘. Click
        on writer Name, enter initial. Click ‗OK

     3. The screen will show e.g. HTLV280499#31.ssn highlighted, click ‗OK‘.

     4. Click on 5 th square from the left: ‗Profile Include‘, click on ‗DetectHTLV#0. prf‘,click ‗OK‘.

     5. Click on 7th square from the left: ‗Sample programming‘.
        First click on ‗Clear sample rack‘, then click on sample code, and enter lab #, and load
        specimens onto specimen rack at the same time. Also load second half of the rack with dilution
        tubes.
        Once finished, double click on ‗Small Rack‘ icon in top left corner, ―X‘s should appear next to
        all lab #s,click ‘done‘.

     6. The screen will show plate format (# of wells needed). Take out the required # of strips and
        wells. Load plate onto the left side in the P-Lab. Click on ‗done‘.

     7. Click on 8th square from the left: ‗Setup Entry‟. Click on pulldown arrow under ‗Lot‖.. If it
        is a new lot, Click on ‗Change Date‘ and specify kit expiration date if needed. Click ‗OK‘.

     8. Click on last square ‗Start Session‘, ‗Save document: using the new file…‘will appear, click
        ‗OK‘.

     9. Profile-vial locations for controls or standards and reagents‘ will appear. Right click on
        each colored position, will show where to load each diluent, controls, conjugate, substrate and
        stop solution. Click ‗continue‘, and wait for instrument initialization.

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 82
                                                                          Policy # MI/SER/09/ v04                         Page 3 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Human T-Lymphotropic Virus
                                                                                         Type 1 (HTLV-1) EIA for
                                                                                         TGLN Call Back Only

                       Position          Reagent & Controls
                        R3               Sample Diluent
                        R4               Conjugate
                        R5               Substrate
                        R6                Stop Solution
                         1               Negative Control
                         2               Positive Control


                      Prepare Fresh (same Day) Conjugate as follows (always make 1 extra
                      strip):
                      Number of Strips 40X conjugate(ul) Conjugate Diluent(ml)
                              1              30                  1.2
                              2              50                  2.0
                              3              80                  2.9
                              4             100                  3.9
                              5             130                  4.9
                              6             150                  5.9
                              7             180                  6.8

                       Prepare Fresh (Same Day) Substrate as follows (always make 1 extra
                      strip):
                       Number of Strips   TMB Reagent(ml) Peroxide Reagent(ml)
                              1              0.2                   1.0
                              2              0.4                   2.0
                              3              0.6                   3.0
                              4              0.8                   4.0
                              5              1.0                   5.0
                              6              1.2                   6.0
                              7              1.4                   7.0

     10. Click on 2 nd square from the left‘ Maintenance‘. Click on ‗Start‘ to start up 'Self Test'.
         Machine will do all checks, then asks: ‗Print Self Test report‘, click on ‗Yes‘. Staple printout
         with day list.

     11. Click on „Daily Maintenance‟: click on ‗Fill syringes‘, check for bubbles in syringes, and if
         no bubbles present, click ‘No‘.

     12. Click on ‘Fill lung‘, screen will ask you to open lid. Open lid, and click on ‗OK‘. Water level
         should fill up to just under the lower black line. Click 1-2 times to add 250 uL of water each
         time to raise the level to just above the lower black line. Close lid and click on ‗OK‖.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 83
                                                                          Policy # MI/SER/09/ v04                         Page 4 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                Subject Title: Human T-Lymphotropic Virus
                                                                       Type 1 (HTLV-1) EIA for
                                                                       TGLN Call Back Only
     13. Click on ‗Buffer 1 prime‘, listen to the noise to make sure that the vacuum is working.

     14. Click on 3 rd square ‗Tips‖ if need to replace pipette box.

     15. Click on 6th square from the left to ‗execute ‗the run, will ask to put sample rack in, click
         ‗OK‘. The P-lab will start processing. Click on ‗Time chart‖ to see what stages the testing is
         at.

     16. After assay is finished, ―Session terminated‖ will appear on screen, click ‗OK‘.

     17. Click on ‗Maintenance, and click on ―Endwork‖. Will ask to fill in buffer 2 with DH2O, open
         lid, and make sure enough DH2O in the bottle, also change buffer 1 bottle to DH2O bottle.
         Click ‗OK‘. The machine is rinsing the tubing with DH2O. Next will ask to ‗empty waste‘,
         press on ―Black Button‖ on the side of the machine, and hold it until almost all fluid is drained
         off and bubbles appear.
         Empty waste. Click ‗OK‘. ‗Empty tips‘, click ‗OK‘.‘Don‘t forget to shut off P-Lab ‗, click
         ‗OK‘.

     18. Click on left top corner ‗File‘, and ‗Exit‘, will ask ‗exit from P lab Processor?‘ Click ‘Yes‘.

     19. Click on 4th box from the top: ‗Open Results‘. Click on ‗New Result‘. Then click on you
         session and ‗OK‘.

     20. Click on 3rd box from the left, ‗report‘, click on ‖DetectHTLV‘, and also the square with the
         ‘printer‘. Report will be printed out.

     21. Close all windows by clicking on ‗X‘ at top right corner three times.

     22. Turn off hard drive, monitor, P-lab and printer.

     Validation

     23. Absorbance of the blank must be< or = 0.100.

     24. The Positive Control mean must be  or = 0.800.

             If the value is less than 0.80, the run must be repeated

     25. Cut off=NCx (negative control mean)+0.150



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 84
                                                                          Policy # MI/SER/09/ v04                         Page 5 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Human T-Lymphotropic Virus
                                                                                         Type 1 (HTLV-1) EIA for
                                                                                         TGLN Call Back Only

     Interpretation of Results:

     1. Absorbance value is  cut off: reported as Negative.
     2. Absorbance value is  or cut off: send to PHL for confirmation.


B. Using NexGen System with West Nile Virus IgM and HTLV combined panel

     1.      Click or touch Monitor on the left column.

     2.      Click or touch ―Manual Controls‖ tab on the top.

     3.      Click or touch ―1‖ Click or touch ―Manual Controls‖ tab on the top.

     4.      Click or touch ―1‖ on the ―Carousel Control‖ section. The segment will move to the front
             and the number is shown in the centre of the ―Carousel Control‖ section.

     5.      Remove the segment.

     6.      Repeat Step 4 for segments 2, 3, 5, 6 and 7.

     7.      Click or touch ―Close‖ on the bottom right corner to close down the ―Monitor‖ session.

     8.      Go to ―Session Builder—Create or modify ELISA session for GexGen‖ and click or touch
             screen. Select panel ―Focus WestNile_PlateA_Adaltis_HTLV_PlateC‖ from the ―Select
             panel or Session‖ field.

     9.      Click or touch ―Next‖ at bottom right of screen.

     10. Click or touch space beside Sample Code. Scan specimens. Repeat until all specimens have
         been scanned.

     11. Click or touch ―Test Name‖ to select all the samples for both WNV IgM and HTLV. Click
         to touch to deselect specimen that does not have to be run.

     12. Click or touch ―Next‖ at bottom right of screen.

     13. To load specimens, follow the ―WorkList‖ table on the ―Session Report‖ on screen and load
         specimen tubes.


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 85
                                                                          Policy # MI/SER/09/ v04                         Page 6 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Human T-Lymphotropic Virus
                                                                                         Type 1 (HTLV-1) EIA for
                                                                                         TGLN Call Back Only

     14. Click or touch ―Next‖ at bottom right of screen. ―I want to build the created session and run
         immediately.‖ Click or touch ―Finish‖ at bottom right of screen. ―Initializing instrument‖
         appears on screen.

     15. Click or touch the sector to load. Load the dilution tubes accordingly. The carousel will
         move to the correct position. Place the sector through the front opening onto the carousel.
         Repeat until all dilution tubes and sectors are loaded. Close door.

     16. Continue with Step 19 in the NexGen general instruction.

C. DETECT-HTLV™ Manual Method

    Procedure:

     1. Remove microplate, reagents and buffer solution from the refrigerator and allow them to come
        to room temperature ( app. 30 minutes).Remove strips from the pouch just prior to use and
        return unused strips to pouch ,seal pouch and return pouch to refrigerator.

     2. Label dilution tubes with sample #s.Add 500 ul of diluent in dilution tube for each sample; add
        10 ul of sample to the corresponding dilution tube. Mix well.

     3. Placement of Controls on microplate:
        A1-Reagent Blank
        B1-Negative Control
        C1-Negative Control
        D1-Negative Control
        E1-Positive Control
        E2-Positive Control
        E3-First sample
        E4-Second sample

     4. Add 100 ul of sample diluent to A1 for use as Blank Control

     5. Add 100 ul of negative control each to B1, C1 and D1.

     6. Add 100 ul of Positive Control each to E1 and F1.

     7. Add 100 ul of diluted sample to the assigned well. Repeat the same process for each sample to
        be tested.

     8. Incubate at room temperature for 30 minutes.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 86
                                                                          Policy # MI/SER/09/ v04                         Page 7 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Human T-Lymphotropic Virus
                                                                                         Type 1 (HTLV-1) EIA for
                                                                                         TGLN Call Back Only

     9. Prepare diluted conjugate as follows (prepare the exact # of strips needed):
        Number of strips       Volume of 40x Conjugate          Volume of Conjugate Diluent
             1                             30ul                               1.2ml
             2                             50ul                               2.0ml
             3                             80ul                              3.2 ml

     10. Discard plate content into a discard container. Fill a squeeze bottle with buffer solution and fill
         each well with buffer solution. Discard the plate content, and re-fill with buffer solution.
         Repeat this process 5 times. Try not to create lots of bubbles during washing. After last wash
         and decant content, try to get rid of all the liquid in the well by tapping the upside down plate
         sharply on clean dry paper towels.

     11. Dispense 100 ul of diluted Peroxidase Conjugate into each well. Incubate for 30 minutes at
         room temperature.

     12. Prepare diluted substrate as follows(prepare the exact # of strips needed):
         Number of strips       Volume of TMB reagent          Volume of Peroxide Reagent
               1                           0.2 ml                             1.0ml
               2                           0.4 ml                             2.0ml
               3                           0.6 ml                             3.0ml

     13. Repeat step 9.

     14. Dispense 100 ul of freshly prepared Substrate into each well. Incubate for 30 minutes at room
         temperature.

     15. Add 100 ul of Stop Solution to each well.

     16. Read the plate in Syva Reader at 450 nm.Turn Syva Reader on (ON/OFF button at the back of
         reader, just above the cord).

             a. Press ―Enter‖ x3 until‖ Plate Read‖ is displayed.
             b. Press‖ Enter‖- will show ―Single wavelength‘. If ―Dual Wavelength‖ is displayed, press
                 ―Option‖ button on top right corner, then will display ―Single Wavelength‖.
             c. Press ‗Enter‘-will show ―Reading WL 450nm‖.
             d. Press ‗Enter‘-will show ―Insert Plate , Press Start,‖
             e. Insert plate, press start. Plate will be read and will print out results from printer sitting
                 on top of the Reader.
           Turn off Syva Reader.


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 87
                                                                          Policy # MI/SER/09/ v04                         Page 8 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Human T-Lymphotropic Virus
                                                                                         Type 1 (HTLV-1) EIA for
                                                                                         TGLN Call Back Only

     Validation:

      Check OD readings on print out sheet:
     1. Absorbance of Blank must be <0.100.

     2. Subtract Blank OD reading from each of the NC OD reading:

          NCx= ∑(NC1 -Blank )+(NC2-Blank) + (NC3-Blank)/3
          Absorbance of the individual Negative Control must be <or =0.150.If one value is outside this
          range, discard this value and recalculate the mean. If two values are outside this range, the run
          must be repeated.

     3. Subtract Blank OD reading from each of the PC OD reading:
        PCx=∑(PC1-Blank)=(PC2-Blank)/2

          The positive mean must be = or >1.400.If the value is less than 1.400, the run must be
          repeated.

     4. Cut-Off=NCx + 0.150


     Interpretation:

     Subtract Blank OD reading from each sample OD reading:
     Negative: < Cut off value
     Positive: >or = Cut Off value, reported as ―send to PHL for further testing‖. Report as
     ―Indeterminate‖ in TGLN web site.

IV        Reporting

          Positive Report:               "POSITIVE"

          Negative Report:               "Negative"




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 88
                                                                          Policy # MI/SER/09/ v04                         Page 9 of 9
                     Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Human T-Lymphotropic Virus
                                                                                         Type 1 (HTLV-1) EIA for
                                                                                         TGLN Call Back Only

V.        Quality Control

          Two positive and three negative controls must be included with each run.

          Run Virotrol I as external control (Do not dilute) with each new lot.

          All positive specimens are sent to PHL for confirmation.

          If any control fails, the test is invalid. Withhold patients‘ results and inform Senior/Charge
          technologists. Repeat testing if warrented.

          Alternative to proficiency testing:
          Proficiency testing is currently not available, therefore, two specimens are sent to PHL every 6
          months (January and July) for verification. The PHL reports are filed in the Alternative
          Proficiency Testing binder.

VI        Reference

          Package insert from DETECT-HTLV, EIA Test Kit for the Detection of
          Antibodies to Human T-Lymphotropic Viruses (HTLV) Types I and II.
          Adaltis, rev.06/02.

          Package insert from DETECT -HTLV™,RTD-902B/900B,Rev 11/03.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 89
                                                                          Policy # MI/SER/10/ v04                        Page 1 of 2
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Infectious Mononucleosis
                                                                                         Heterophile Antibodies
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: July 23, 2006
                                                                          Annual Review Date: January 10, 2011

                                            Infectious Mononucleosis Heterophile Antibodies

I.        Introduction

          The MONOSPOT LATEX slide test is a latex particle agglutination test for in vitro qualitative
          detection of infectious mononucleosis heterophile antibodies (IgM) in serum or plasma. These
          antibodies appear in the sera of 85 to 90% of patients with infectious mononucleosis within 2
          to 3 weeks after onset of illness.

II.       Specimen Collection and Processing

          5 mL of blood is collected in a serum separator tube and separated by centrifugation. The tube
          is refrigerated until testing. Specimens are stored at -200C after testing and discarded after 3
          months.

III.      Procedure

          i) Reagents:

                    MONOSPOT LATEX Kit:

                    Store refrigerated. Allow the reagent to warm up to RT. Mix well before use.

          ii) Other Materials:

                    Supplied with kit:
                           Test slides
                           Paddle pipettes

          iii) Method:

                1. Dispense 1 drop of the latex reagent onto a labelled oval ring of the test card.

                2. Add 1 drop (50 L) of patients's serum or control to the same ring.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 90
                                                                          Policy # MI/SER/10/ v04                        Page 2 of 2
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Infectious Mononucleosis
                                                                                         Heterophile Antibodies

                3. Mix the latex reagent and serum together and spread to cover the entire area of the ring
                   with the blade end of the paddle pipette.


                4. Immediately rotate the card on the serologic rotator at 100 rpm for 3 minutes.

                5. Observe for agglutination using a light source to aid in visualization.

          iv) Interpretation of Results:

                    Negative:            No agglutination

                    Positive:            Any degree of agglutination

IV.       Reporting

          Positive Report:               "Infectious mononucleosis heterophile antibody: POSITIVE"

          Negative Report:               "Infectious mononucleosis heterophile antibody: NEGATIVE"

V.        Quality Control

          Negative and positive controls must be included with each run and results and kit lot number
          recorded on the tasklist. When opening a new kit, record the lot number in the reagent lot
          number binder. Refer to a senior technologist if control results are outside of limits or for any
          other problems with running or reporting the assay.

          Run external control ( Accurrun 31) with each new lot. Result filed in
          Reagent Lot Binder. . If result is negative, inform Charge/senior technologist for review.

          CAP provides external proficiency testing.

VI.       References

          Manufacturer's package insert: Meridian Diagnostics, Inc., 3471 River Hills Dr., Cincinnati,
          Ohio 45244 U.S.A. 1-513-271-3700.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 91
                                                                          Policy # MI/SER/11/ v04                         Page 1 of 3
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Rapid Plasma Reagin Test
                                                                                 for Syphilis Screening
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: January 31, 2011
                                                                          Annual Review Date: January 10, 2011

                                                                 Syphilis Screening

I.        Introduction

          The NCS Rapid Plasma Reagin (RPR) Card test is a macroscopic non-treponemal flocculation
          test used to detect reagin antibodies.

II.       Specimen Collection and Processing

          Blood is collected (5 mL for adult and 1 mL for neonate) in a serum separator tube and
          separated by centrifugation. The serum is removed to a tube and refrigerated until testing.
          Specimens are stored in the refrigerator for 3 months after testing. A request for VDRL on
          spinal fluid (CSF) or neonate blood will be sent to PHL for testing.
          Note: RPR must be used for cadaveric donor syphilis testing.

III.      Procedure

          i)        Reagents:

                    RPR reagent kit (NCS Diagnostics Inc.)

          ii)       Other Materials:

                    3 mL dropper bottle
                    Dispensing needle (17 L/drop)
                    RPR test card
                    0.05 mL disposable stirrer pipettes
                    Serological rotator at 100 rpm
                    dH2O

          iii)      Precaution:

                    Refrigerate reagents until required. Warm to RT and mix well before use. To ensure
                    stability, return the antigen suspension to the original glass bottle after testing. The
                    dispenser and needle assembly must be thoroughly washed in dH2O and air dried after
                    use.



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 92
                                                                          Policy # MI/SER/11/ v0                          Page 2 of 3
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Rapid Plasma Reagin Test
                                                                                         for Syphilis Screening


          iv)       Needle Accuracy Check - When New Kit Opened:

                    This procedure is performed to check the needle delivering antigen each time before
                    testing. Using a pipette, deliver 0.5 mL antigen to the dropper bottle. Attach the needle
                    and, holding in a vertical position, count the number of drops delivered in 0.5 mL. The
                    needle is considered satisfactory if 30  1 drops are obtained. If the needle is
                    unsatisfactory, repeat the check. Record the lot number of the newly opened kit in the
                    reagent lot number binder.

          v)        Method:

          1.        Using the stirrer pipette held vertically, dispense one drop (50 L) of serum onto a
                    circle on the test card. Use a fresh stirrer pipette for each sample. Repeat with the
                    control sera.

          2.        Using the flat end of the stirrer pipette spread the sample over the entire area of the test
                    circle.

          3.        Attach the needle to the dropper bottle. Mix the carbon antigen reagent well. Squeeze
                    the dropper bottle and withdraw sufficient reagent into the bottle. Discard the first few
                    drops into the reagent stock bottle and then dispense 1 drop into each circle in a vertical
                    position. Do not mix the sample and the antigen. Rotate the card at 100 rpm for 8
                    minutes.

          4.        Observe for agglutination by two technologists independently.

          vi)       Interpretation of Results:

                    Positive Result:          Any agglutination. Repeat test ---- see senior technologist for any
                                              discrepant results. Send all positive sera to PHL for VDRL and
                                              confirmatory tests. Write on PHL requisition: "RPR positive, please
                                              do confirmatory test"; DO NOT mark prenatal box for serum from
                                              prenatal patients.

                    Negative Result: No agglutination.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 93
                                                                          Policy # MI/SER/11/ v04                         Page 3 of 3
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Rapid Plasma Reagin Test
                                                                                          for Syphilis Screening


IV.       Reporting

          Positive Result:               Enter in LIS as "TO PHL". VDRL send-out test is ordered reflexively.
                                         Send to PHL next day.

          Negative Result:               Negative

V.        Quality Control

          Strongly reactive, weakly reactive and non-reactive control sera are included in each run. If
          controls are not working or the antigen is not falling cleanly from the needle, perform a needle
          check as outlined in the method.

          Record control results and kit lot number on the task list. Run external control (Accurun 156)
          with each new lot When opening a new kit, record the lot number, needle check and external
          control results in the reagent lot binder.
          Refer to a senior technologist if control results are outside of limits or for another problems
          with running or reporting the assay.

          CAP provides external proficiency testing.

VI.       Reference

          Manufacturer's package inserts (RPR Card Test, NCS Diagnostics Inc., 130 Matheson Blvd. E.
          Mississauga, Ontario L4X 1Y6).




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 94
                                                                          Policy # MI/SER/12/ v04                        Page 1 of 6
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Varicella-Zoster Virus IgG
                                                                                         VIDAS System
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: Feb 18, 2008
                                                                          Annual Review Date: January 10, 2011

                                                           Varicella-Zoster Virus IgG
I.        Introduction

          Varicella-Zoster IgG assays is used for evaluating patient's immune status to Varicella-Zoster
          viruses infection. The VIDAS is an enzyme-linked fluorescent immunoassay (ELFA) utilizing a
          virus coated Solid-Phase-Receptacle (SPR) to which antibody in serum binds. Anti-Human IgG
          conjugated with Alkaline Phosphatase reacts with substrate, 4-Methylumbelliferyl Phosphate to
          form a fluorescent product. All necessary reagents and test serum are contained in the VZG
          reagent strips.

II.       Specimen Collection and Processing

          Blood (5 mL) is collected in a serum separator tube and separated by centrifugation. The serum is
          removed to a vial and refrigerated until testing. Specimens are stored at -20oC after testing.

III.      Procedure

          Reagents

          VIDAS Varicella-Zoster IgG test kit:
               VZG Reagent Strips
               VZG SPRs
               Standard
               Positive Control
               Negative Control

          Other materials:
                 Pipettor 100 uL

          Method

          1.        Bring test kit and serum samples to room temperature. Check that lot number of kit
                    matches lot currently in use (as posted on VIDAS instrument). If lot number does not
                    match, ensure that all kits/test strips for the posted lot number have been used.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 95
                                                                          Policy # MI/SER/12/ v04                        Page 2 of 6
                      Microbiology Department
Policy & Procedure Manual
Serology Manual                                                           Subject Title: Varicella-Zoster Virus IgG
                                                                                         VIDAS System

                    Enter new lot data by inserting bar code card found in kit into tray, load tray into Section
                    ‗A‘. Press ‗Master Lot Menu‘,‘ Read Master Lot‘ and section ‗A‘. The machine will move
                    the tray and read the card. At the end, press ‗Master Lot Menu‘, use ↓ to ‗List Master Lot‘.
                    Press that button, and also ‗VZG‘, will show three lots #. Check that one of these includes
                    the new lot.

                    Post card of new lot currently in use on VIDAS along with the date.

          2.        Label MSG/VZG strips as follows:
                                s (Standard)
                                s (Standard)
                                C1 (Positive Control)
                                C2 (Negative Control)
                                5
                                6 etc. up to 30

          3.        Pipette 100 uL of Standard, Control or serum into
                    the specimen well of the corresponding VZG strips.
                    Check for adequate sample level and remove any bubbles.

          4.        At VIDAS instrument, start at Main Menu.

                    To program a run with Standards( Alarm will alert the expiration of
                    standards):

                     a. Load strips(containing 100 ul of each standards, controls, or samples) and tips into
                        Mini Vidas.

                     b. Press ‗Status Screen‖- will show ‗A‘ and ‗B‘ available.

                     c. Press ‗A‘.

                     d. Section ‗A‘ appears with 1,2,3,4,5,6.

                     e. Press‘1‘(Position 1), then press ‗S‘ for Standard.

                     f. Standard number (1-4) appears, press ‗1‘,will show ‗S1‘,‘Enter‘.

                     g. Press ‗2‘(position 2), then press ‗S‘ for Standard.



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 96
                                                                          Policy # MI/SER/12/ v04                        Page 3 of 6
                      Microbiology Department
Policy & Procedure Manual
Serology Manual                                                           Subject Title: Varicella-Zoster Virus IgG
                                                                                         VIDAS System

                     h. Standard number (1-4) appears, press ‗1‘ (not 2) ,will show ‗S1‘,‘Enter‘.
                     i. .Press ‗3‘ (position 3), then press ‗C‘ for Control.

                     j. Control number (1-4) will appear, press ‗1‘, will show ‗C1‘,‘Enter‘.

                     k. Press ‗4‘ (position 4), then press ‗C‘ for Control.

                     l. Control number (1-4) will appear, press ‗2‘, will show ‗C2‘,‘Enter‘.

                     m. Press ‗5‘ (position 5), then press ‗Sample ID‘.

                     n. Use arrow to move cursor to ‗D‘, press button connected to that line (Last
                        button).Then press # on the keyboard to enter all the numbers.‘ Enter‘.

                     o. Press ‗6 (position 6), then press ‗Sample ID‘.

                     p. Use arrow to move cursor to ‗D‘, press button connected to that line (Last
                        button).Then press # on the keyboard to enter all the numbers.‘ Enter‘.

                     q. Press ‗Start‘. Once check has been completed and ‗OK‘, green light will stay on,
                        and screen will show the time the testing will be finished.

                     r. Press ‘B‘ for next module.

                     s. Press ‗1 (position 1), then press ‗Sample ID‘.

                     t. Use arrow to move cursor to ‗D‘, press button connected to that line (Last
                        button).Then press # on the keyboard to enter all the numbers.‘ Enter‘.

                     u. Do the same for the rest of the positions.

                     v. Press ‗Start‘. Once check has been completed and ‗OK‘, green light will stay on,
                        and screen will show the time the testing will be finished.

                     w. Once the testing is finished,‘ Unload‘ will appear. Unload strips and tips. Check
                        strips that show ‗NEGATIVE‘ for inoculums.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 97
                                                                          Policy # MI/SER/12/ v04                        Page 4 of 6
                      Microbiology Department
Policy & Procedure Manual
Serology Manual                                                           Subject Title: Varicella-Zoster Virus IgG
                                                                                         VIDAS System

               To program a run with without Standard:

                 a. Load strips (containing 100 ul of each controls, or samples) and tips into Mini Vidas.

                 b. Press ‗Status Screen‖- will show ‗A‘ and ‗B‘ available.

                 c. Press ‗A‘.

                 d. Section ‗A‘ appears with 1, 2, 3, 4, 5, 6.

                 e. Press ‗1‘(position 1), then press ‗C‘ for Control.

                 f. Control number (1-4) will appear, press ‗1‘, will show ‗C1‘,‘Enter‘.

                 g. Press ‗2‘ (position 2), then press ‗C‘ for Control.

                 h. Control number (1-4) will appear, press ‗2‘, will show ‗C2‘,‘Enter‘.

                 i. Press ‗3‘ (position 3), then press ‗Sample ID‘.

                 j. Use arrow to move cursor to ‗D‘, press button connected to that line (Last button).
                    Then press # on the keyboard to enter all the numbers.‘ Enter‘.

                 k. Do the same for the rest of the positions.

                 l. Press ‗Start‘. Once check has been completed and ‗OK‘, green light will stay on, and
                    screen will show the time the testing will be finished.

                 m. Press ‘B‘ for next module.

                 n. Press ‗1 (position 1), then press ‗Sample ID‘.

                 o. Use arrow to move cursor to ‗D‘, press button connected to that line (Last
                    button).Then press # on the keyboard to enter all the numbers.‘ Enter‘.

                 p. Do the same for the rest of the positions.

                 q. Press ‗Start‘. Once check has been completed and ‗OK‘, green light will stay on, and
                    screen will show the time the testing will be finished.

                 r. Once the testing is finished,‘ Unload‘ will appear. Unload strips and tips. Check strips
                    that show ‗NEGATIVE‘ for inoculums.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 98
                                                                          Policy # MI/SER/12/ v04                        Page 5 of 6
                      Microbiology Department
Policy & Procedure Manual
Serology Manual                                                           Subject Title: Varicella-Zoster Virus IgG
                                                                                         VIDAS System

          Interpretation of Results

          Specimens with test values greater than 0.9 (MEA/VZ) are considered positive. Test values
          ranging from 0.6-0.9 are equivocal.
          Samples with test values less than 0.6 (MEA/VZ) are considered negative.


IV.       Reporting

          Positive:                      Varicella-Zoster antibody:                Positive

          Equivocal:                     Varicella-Zoster antibody:                Equivocal

          Negative:                      Varicella-Zoster antibody:                Negative


V.        Quality Control

          Standard: RFV must be greater than or equal to RFV range posted on VIDAS instrument.

          Positive and Negative Control: Test Values must be within ranges posted on VIDAS.

          Quality Control ranges posted on VIDAS may change from lot to lot, therefore it is essential that
          the lot number of the test kit in use corresponds to that of the posted values.

          Refer to a senior technologist if control results are outside of limits or for any other problems with
          running or reporting the assay.

          Run external control (pooled positive sera) with each new lot. Results filed in Reagent Lot
          Binder. If result is negative, the run is invalid. Inform Charge/senior technologist, and repeat
          testing.

         CAP provides external proficiency testing.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 99
                                                                          Policy # MI/SER/12/ v04                        Page 6 of 6
                      Microbiology Department
Policy & Procedure Manual
Serology Manual                                                           Subject Title: Varicella-Zoster Virus IgG
                                                                                         VIDAS System

Monthly:

      Run QCV :

      1. Press ‗Status Screen‘.
      2. Press section‗A‘.
      3. Press position ‗1‘.
      4. Press ‗Assay‘ and ‗Select Assay‘.
      5. Press ‗QCV‘.
      6. Press ‗Enter‘.
      7. Press position ‗2‘, will show‘QCV‘ on screen, ‗Enter‘.
      8. Repeat step # 7 for positon 3-6.
      9. Load 6 strips and tips from ‗QCV kit‘.
      10. Press ‗Start‘.
      11. Repeat step 1 to 9 for Section ‗B‘.
      12. Check values: TV1 >= value stated on box
                         R3 >= 4100 RFU
      13. Staple print out sheets and file in MiniVidas binder, initial ‗ Monthly QC‘.

      Clean Lens:

      1. Open the left side of section A where tips are loaded all the way down.
      2. Use device provided from Vidas to get rid of any dust on the lens by pumping air onto the
         lens.

VI.       Reference

          Manufacturer's Package Insert.
          MINI-VIDAS System Operational Manual.
          bioMerieux-Vitek, Inc.
          Missouri, USA 64042-2395




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 100
                                                                          Policy # MI/SER/13/v04                          Page 1 of 6
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: West Nile Virus IgM EIA Test
Issued by: LABORATORY MANAGER                                             Original Date: Feb 19, 2007
Approved by: Laboratory Director                                          Revision Date: Feb 19, 2008
                                                                          Annual Review Date: January 10, 2011

                                                        West Nile Virus IgM EIA Test

I.        Introduction

       The Focus Technologies Flavivirus (West Nile) IgM capture ELISA is intended for
       qualitatively detecting IgM antibodies to flaviviruses(West Nile) in human serum.
       In conjunction with the Focus Technologies Flavivirus (West Nile) ELISA IgG,
       the test is indicated for testing persons having symptoms of arbovirus infection,
        as an aid in the presumptive diagnosis of flavivirus infection.

II.       Specimen Collection and Processing

       a. Approximately 10 ml of blood is collected in a red-topped tube.
       b. Centrifuge the tube at 2000 rpm for 10 minutes.
       c. Transfer serum into 2 ml storage tube and store in designated rack in 4 C fridge.

III.      Procedure

          P-Lab
          i.    Reagents:

               a. Wash Buffer (100 mL) 10X: Prepare working buffer (1X) by adding 100 ml of 10X
                  Wash Buffer to 900 mL of Distilled H2O. Mix well. Labeled and dated.
               b. West Nile Antigen (2 vials): Add exactly 8 mL of the Sample Diluent to 1 vial of
                  Antigen. ( Do not use Distilled H2O). Allow the antigen to rehydrate at room
                  temperature for 1 hour prior to use- the antigen must be completely dissolved before
                  use. Store the remaining Antigen at 2 to 8oC for up to 60 days following reconstitution.
                  Avoid storing the reconstituted Antigen at room temperature: remove from 2 to 8oC,
                  withdraw the amount needed immediately, and return the rest immediately to 2 to
                  8oC.Do not freeze.
               c. 1 vial of Positive Control,0.3 mL
                  1 vial of Negative Control, 0.3 mL
                  1 vial of Cut-off Calibrator (Correction factor-CF-stated on vial) 0.3 mL
               d. Anti-flavivirus Conjugate, 16 mL
               e. Substrate Reagent: 2 vials of Tetramethylbenzidine (TMB) and Hydrogen Peroxide in
                  buffer
               f. Stop Reagent, 16mL
               g. IgM Capture Wells,96 wells
               h. Sample Diluent, 112 mL

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 101
                                                                          Policy # MI/SER/13/v04                          Page 2 of 6
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: WNV IgM EIA Test

          ii.       Procedure:

           1.       Before beginning EIA assay:
                     Allow all components to reach room temperature. Remove 6 strips and refrigerate
                       the rest of the strips right away. This run will include 5 controls and 43 specimens.
                     If newly reconstituted, allow re-constituted Antigen to dissolve at room temperature
                       for 1 hour before use.
                     Fill up P-Lab wash bottle with working Wash buffer. Place bottle in ‗Buffer #1‘
                       position in P-Lab.

           2.       Turn hard drive and monitor. Wait for software to be fully loaded before turning the P-
                    Lab and printer ‗ON‘. Put pencil in the hole.

           3.       Type in ‗qw‘, tab, ‗qw‘, enter.

           4.        Click on ‗New‖, type in assay name & date performed, e.g. WNV IgM 040530 (tested
                    2004 May 30). Note session # e.g.‘31‘.Click on writer Name, enter initial. Click ‗OK.

           5.       The screen will show e.g. WNV IgM4#31.ssn highlighted, click ‗OK‘.

           6.       Click on 5th square from the left: ‗ Profile Include‘, click on
                    ‗Focus WNV IgM #0.prf‘, click ‗OK‘.

           7.       Click on 7th square from the left : ‗Sample programming‘.
                    First click on ‗Clear sample rack‘, then click on‘ ID Range‘, and scan barcode of the
                    lab # or enter in manually and load specimens onto the specimen rack at the same time.
                    Also load dilution 43 dilution tubes into the second half of the rack.
                    Once finished, double click on ‗Small Rack‘ icon in top left corner, ―X‘s should appear
                    in WNV IgM column, click ‘done‘.

           8.       The screen will show plate format (# of wells needed). Load plate onto the left side.
                    Click ‗done‘

           9.       Click on 8th square from the left: ‗Setup Entry‟. Click on‘ Lot # ↓‘, highlight correct
                    lot #. Click ‗OK‘. (Or Enter new lot #. Click on ‗Change Date‘ and change kit
                    expiration date if needed-always change year first.). Click ‗OK‘.

           10.      Click on last square ‗Start Session‘, click ‗OK‘. ‗Save document…‘, click ‗OK‖.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 102
                                                                          Policy # MI/SER/13/v04                          Page 3 of 6
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: WNV IgM EIA Test

           11.      Profile-vial locations for controls or standards and reagents‘ will appear. Right
                    click on each colored position, will show where to load each diluent, controls,
                    conjugate, substrate, and stop solution including amount needed. Click ‗continue‘, and
                    wait for initialization.
                                     Position        Reagent & Controls
                     R3              Sample Diluent
                     R4              Re-constituted Antigen
                     R5              Conjugate
                     R6              Substrate
                     R7              Stop solution
                     1                Negative Control
                     3                Cut off Calibrator
                     5                Positive control
                     2                Dilution cup for Negative Control
                     4                Dilution cup for Calibrator
                     6                Dilution cup for Positive Control

           12.      Click on 2 nd square from the left‘ Maintenance‘ Click on ‗Self Test‘, then ‗Start‘.
                    Machine will do all checks, then asks: ‗Print Self Test report‘, click on ‗Yes‘. Staple
                    printout with day list.

           13.      Click on 2 nd square from the left‘ Maintenance‘ Click on ‗Self Test‘, then ‗Start‘.
                    Machine will do all checks, then asks: ‗Print Self Test report‘, click on ‗Yes‘. Staple
                    printout with day list.

           14.      Click on „Daily Maintenance‟: ‗Open front door‘, click ‗OK‘. Click on ‗ Fill
                    syringes‘, check for bubbles in syringes, and if no bubbles present, click ‘No‘.

           15.      Click on „Daily Maintenance‟: ‗Open front door‘, click ‗OK‘. Click on ‗Fill
                    syringes‘, check for bubbles in syringes, and if no bubbles present, click ‘No‘.

           16.      C lick on ‘Fill lung‘, screen will ask you to open lid. Click on ‗OK‘.Water level should
                    fill up to just under the lower black line. Click 1-2 times to Add 250ul of water each
                    time to raise the level to just above the black line (DO NOT OVERFILL). Close lid
                    and click on ‗OK‖.

           17.       Click on ‗Buffer 1 prime‘, listen to the noise to make sure that the vacuum is working
                    ,check to make sure that the level of fluid is actually going down. ‗Close front door‘,
                    Click ‗OK‘.

           18.      Click on 3 rd square ‗Tips‖ if need to replace pipette box. Click on 6th square from the
                    left to ‗execute ‗the run, will ask to put sample rack in, click ‗OK‘. The P-lab will start
                    processing.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 103
                                                                          Policy # MI/SER/13/v04                          Page 4 of 6
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: WNV IgM EIA Test

           19.      Click on ‗Time chart‖ to see what stages the testing is at.

           20.      After assay is finished, ―Session terminated‖ will appear on screen, click
                    ‗OK‘.
           21.      Click on 2nd square‘ Maintenance and ‗Daily Maintenance‘. Click on ―Endwork‖. Will
                    ask to fill in buffer 2 with DH2O, open lid, and make sure enough DH2O in the bottle.
                    Change buffer 1 bottle to DH2O bottle. Click ‗OK‘. The machine is rinsing the tubing
                    with DH2O. Next will ask to ‗empty waste‘, press on ―WASTE‖ button on the side of
                    the machine, and hold it until almost all fluid is drained off and bubbles appear. Empty
                    waste. Click ‗OK‘. ‗Empty tips‘, click ‗OK‘. ‗Don‘t forget to turn off machine‘, click
                    ‗OK‘.
           22.      Click on left top corner ‗File‘, and ‗Exit‘, will ask ‗ exit from WB Processor?‘, click‘
                    Yes‘.
           23.      Click on 4th box from the left : ‗Open Results‘,‖ new result‘. Click on your session
                    and ‗OK‘.

           24.      Click on 3rd box from the left, ‗ report‘, Choose ‘Focus WNV IgM‘ and click on 5th
                    square with the ‘printer‘, Report will be printed out.

           25.      Close all windows by clicking on ‗X‘ at top right corner.

           26.      Click on ‗Start‘ on lower left corner,‘ Shut Down‘ and ‗OK‘.Turn off
                    P-Lab, hard drive, monitor, and printer.

          I. Manual Method:

               1. Label a set of 12x75 mm glass tubes as follows: Blank, Pos,Neg, Cal,7,8,9,10….,and
                  External Control (when required, e.g. a new lot #).
               2. Add 1 ml of Sample Diluent to all tubes.
               3. Leave 1st tube blank; add 10 ul of Controls, Calibrator and patient samples to
                  corresponding tubes.
               4. Take out the required # of strips (Open bag containing strips when strips reaches room
                  temperature, ~10-15 minutes). Return the unused strips to the bag, and refrigerate
                  immediately.
               5. Change bottle from D H2O to Wash Buffer. Fill all wells with wash buffer, and
                  soak for 5 minutes, do this 3 times.
               6. After washing, remove all wash buffer by inverting the plate onto clean paper towel.
               7. Add 100 ul of diluent to well # 1, 100 ul of Pos to well # 2, 100 ul of Neg to well # 3,
                  100 ul each of Cal to well # 4, 5, and 6. Add 100 ul of specimen to each well, starting
                  from well # 7. Use last well as external
                   Control well when needed.
               8. Cover plate with sealing tape, and incubate for 60 minutes at Room Temperature.
               9. Repeat steps # 5 and #6.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 104
                                                     Policy # MI/SER/13/v04            Page 5 of 6
                        Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                             Subject Title: WNV IgM EIA Test
          10. Add 100 ul of Antigen into each well using an 8-channel pipette, cover plate with
              sealing tape and incubate for 2 hours (120 minutes) at room temperature.
          11. Repeat wash steps # 5 and # 6.
          12. Add 100 ul of Conjugate into each well using an 8-channel pipette, cover plate with
              sealing tape and incubate for 30 minutes at room temperature.
          13. Repeat wash steps # 5 and # 6.
          14. Add 100 ul of Substrate into each well using an 8-channel pipette, cover and incubate
              for 10 minutes at room temperature.
          15. Add 100 ul of Stop Reagent into each well using an 8-channel pipette.
              In antibody-positive wells, color should change from blue to yellow.
          16. Read the plate in spectrophotometer at singlel wavelength 450 nm within 1 hour of
              stopping the assay.

          II. Background Subtraction Assay:
                 1. Set up test manually.
                 2. Set up as follows:
                                a. Blank well (Buffer)
                                b. NC
                                c. PC
                                d. Cal 1
                                e. Cal 2
                                f. Cal3
                                g. Specimen with Antigen
                                h. Specimen with Specimen diluent
                 3. Dilute samples and control: 1:100 in sample diluent (10 ul+1000ul)
                 4. Soak wells for 5 minutes with 1X Wash.
                 5. Add 100 uL of contols,calibrators and specimen into assigned well.
                 6. Incubate for 60 minutes at room temperature.
                 7. Wash 3 times.
                 8. Add 100 uL of Conjugate.
                 9. Incubate for 30 minutes at room temperature.
                 10. Wash 3 times.
                 11. Add 100 uL of Substrate.
                 12. Incubate for 10 minutes at room temperature.
                 13. Add 100 uL of Stop Reagent.
                 14. Read at Syva Reader at 450 nm.

                   Calculation:
                   1. Index for Controls: Control OD/cut off OD.
                   2. Index for Patient samples: (Ag OD-SD OD)/cut off OD.

                   Interpretation:
                   Positive : Index >1.10, inform John/Lily/Linda,send to PHL
                   Inderminate due to high background: Index <0.9
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 105
                                                                          Policy # MI/SER/13/v04                          Page 6 of 6
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: WNV IgM EIA Test

     III.      Validation

               The Positive Control index values must be between 1.5 and 3.5.

               The Negative Control index values must be less than 0.8.

               The mean value for the Cut-Off Calibrator must be within 0.100 and 0.700 units. All
               replicate Cut-Off calibrator ODs should be within 0.10 absorbance units from the mean
               value.

               If any of these do not meet specifications, the test is invalid. The assay has to be repeated.

     IV.       Interpretation

               Index value = Optical Density of Sample/the mean Optical Density of the Cut-Off
                             Calibrators

               Positive: >1.1 Index value
               Equivocal: 0.9-1.1 index value
               Negative : < 0.9 Index value

     V.        Reporting

               Negative for WNV IgM
               Do Background Substraction Assay if Positive or Equovical for WNV IgM

     VI.       Quality Control

               One positive and one negative control must be included with each run.
               Run Accurun 165 (WNV IgG/IgM) as external control (Do not dilute) with each new lot.
               Do background Subtraction Assay for all positive or equivocal specimens.
               If result remains the same, send to PHL for confirmation.

               Alternative to proficiency testing:
               Proficiency testing is currently not available, therefore, two specimens are sent to PHL
               every 6 months (January and July) for verification. The PHL reports are filed in Alternate
               Proficiency Testing binder.

     VII.      Reference

               Package insert of West Nile Virus IgM Capture DxSelect™ by Focus Diagnostics,
               Cypress, California 90630, USA.
               Date written: 27/07/2005.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 106
                                                                                   Policy # MI/SER/15/01/v04                               Page 1 of 1
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                    Subject Title: Appendix I
                                                   Serology Test Schedule
Issued by: LABORATORY MANAGER               Original Date: March 14, 2001
Approved by: Laboratory Director            Revision Date: May 20, 2005
                                            Annual Review Date: November 18, 2009
                        APPENDIX I - SEROLOGY TEST SCHEDULE

EBV Serology......................................................................................................................... Thursday
HBsAg ........................................................................................................................................... daily
HBsAg confirmation ........................................................................................................ when needed
HBsAb ........................................................................................................................................... daily
HBcAb-Total ................................................................................................................................. daily
HBcAb-IgM ...................................................................................................................... when needed
HAV-IgM ...................................................................................................................................... daily
HBeAg / HBeAb .......................................................................................................................... .daily
Monospot IM heterophile Ab ................................................................................................. same day
Rubella Ab ..................................................................................................................................... daily
Varicella-Zoster Ab .................................................................................................STAT or Thursday
RPR for Syphilis .................................................................................................. STAT or Wednesday
HCV Ab ......................................................................................................................................... daily
CMV IgG Ab ................................................................................................................................. daily
HIV Ab .......................................................................................................................................... daily
HTLV Ab.................................................................................................................STAT or Thursday
*          Stat testing may be required where clinically justified.
#
           All routine pre-Bone Marrow Transplant serology screening will be completed by Friday
           afternoon weekly (except RPR for Syphilis for samples received Thurs. pm or Fri.) Friday
           afternoon print an LIS report for 8TSC template. For all BMT patients/donors appearing on
           the 8TSC printout, check that they were faxed successfully. To maintain confidentiality, results
           for HIV testing will be reported using "Transplant Screen". These results will be printed in-lab
           and sent to the PMH BMT office daily by hospital mail in an envelope marked
           "CONFIDENTIAL".(FAX number 926-6585).

           Occasionally a pre-BMT serology screen will be STAT. The microbiologist will inform the
           laboratory in addition to the requisition being clearly marked STAT. Such specimens will be
           processed within 24 hours of receipt and results will be FAXed back to the BMT office as soon
           as they are available.
                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                          Page 107
                                                                             Policy # MI/SER/15/02/v04                    Page 1 of 2
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                     Subject Title: Appendix II - Send-Outs
Issued by: LABORATORY MANAGER                                                Original Date: March 14, 2001
Approved by: Laboratory Director                                             Revision Date: July 23, 2006
                                                                             Annual Review Date: January 10, 2011

                        APPENDIX II - List of Tests Referred Out to Other Laboratories

          TEST                                                REQUISITION                               DESTINATION

                    1TEST                                               REQUISITION                               DESTINATION

1.       Amoebic serology                                    PHL General                               Serology PHL

2.       Aspergillus precipitins                              PHL General                               Serology PHL

3.       Avian precipitins                                    HICL label/requisition                    Hospital In-Comm Lab
                                                              Specify bird in question                  1 William Morgan Dr.
                                                                                                        Toronto, Ontario
                                                                                                                 M4H 1N6
                                                                                                        Tel. (416) 422-3000

4.        Chlamydial culture*                                 PHL General                               Virology PHL

5.        Diphtheria antitoxin                                Reference Bacteriology                    Special
          toxin                                                                                         Bacteriology
                                                                                                        PHL
6.        Fungal antibodies                                   PHL General                               Serology PHL

7.        Malaria                                             PHL General                               Parasitology PHL

8.        Mycoplasma PCR                                      PHL General                               Bacteriology PHL

9.        Genital Mycoplasma*/                                PHL General                               Mycoplasma PHL
          Ureaplasma culture*

10.       Legionella (tissue)                                 PHL General                               Bacteriology PHL
          Legionella urine antigen                            PHL General                               Serology

11.       Lyme disease Ab                                     PHL General                               Serology PHL

12.      PARASITIC SEROLOGY                                    PHL General                               Serology PHL




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 108
                                                                          Policy # MI/SER/15/02/v04                     Page 2 of 2
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Appendix II - Send-Outs

          TEST                                           REQUISITION                              DESTINATION

13.      PARASITIC SEROLOGY cont‘d
          Only by special request from Topical Diseases' physicians:
         Cystercosis                        CDC 50.34 Rev.09/2002                                      Center for Disease
         Echinococcus                       (must be completed by                                      Control and prevention
         Leishmania                          ref .physician)                                           1600 Clifton Road,N.E.
         Schistosoma                                                                                   Atlanta, Georgia
         Strongyloides                                                                                  U.S.A. 30333
         Toxocara                                                                                      Control and prevention
         Trichinella                                                                                   1600 Clifton Road,N.E
         Trypanosoma                                                                                    Atlanta, Georgia
         Miscellaneous                                                                                 U.S.A. 30333

         Filaria                                       NIH Filaria form                            Laboratory of ParasiticDiseases
                                                      (must be completed by                        National Institute of Allergy &
                                                       ref.physician)                              Infectious Diseases
                                                                                                   National Institutes of Health
                                                                                                   Building 4,Rm 126
                                                                                                   Bethesda,Maryland
                                                                                                   U.S.A. 20892
14.       Tetanus antitoxin titre                             Reference Bacteriology                  Special Bact. PHL

15.      Toxoplasma IgG Total                                 PHL form 97-44(08/99)                     Serology PHL
         Toxoplasma IgM

16.       VDRL (CSF)                                          PHL General                               Serology PHL

17.       Electron microscopy                                 PHL General                               Virology PHL
                                                                                                        Tel. (416) 422-3000

18.       Parvovirus Ab IgG, IgM                              PHL General                               Serology PHL

19.       Histoplasma Antigen                                                                       MiraVista Diagnostics
          in Urine                                                                                  4444 Decatur Blvd., Suite 300
                                                                                                    Indianapolis, IN 46241
                                                                                                    1-866-Mira Vista (647-2847)
                                                                                                    Phone: 317-856-2681
20.       Multi-Resistant Organism for                                                       Special Research Microbiology
          Multiple Combination                                                               Room 3043B
          Bactericidal Test                                                                  Children‘s Hospital of
                                                                                             Eastern Ontario

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 109
                                                                          Policy # MI/SER/15/03/v04                        Page 1 of 1
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Appendix III - Shipment of Samples to
                                                                                             ST. JOSEPH'S HOSPITAL
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: May 20, 2004
                                                                          Annual Review Date: January 10, 2011

                APPENDIX III - SHIPMENT OF SAMPLES to ST. JOSEPH'S HOSPITAL

PHL requires that these samples be packaged in a standardized way before they will forward them to
Hamilton for us:

1.        Put sample tube inside plastic bag and place together with requisition into a plastic cylinder.

2.        Place plastic cylinder into a cardboard box, padded with paper towels.

3.        Seal the box with packing tape and label the box as follows:



                                       Public Health Laboratory, 81 Resources Rd.


                                                           DO NOT OPEN


                               Please forward to:


                               Dr. James Mahoney,
                               REGIONAL VIROLOGY LABORATORY,
                               ST. JOSEPH'S HOSPITAL,
                               50 CHARLTON AVE., E.,
                               HAMILTON, ONTARIO
                               L8N2X6




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 110
                                                                          Policy # MI/SER/15/04/v01                        Page 1 of 3
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Appendix IV - Entering Serology
                                                                                         Refer-Out Results
Issued by: LABORATORY MANAGER                                             Original Date: March 14, 2001
Approved by: Laboratory Director                                          Revision Date: January 15, 2004


                     APPENDIX IV - ENTERING SEROLOGY REFER-OUT RESULTS

Refer-out test results for blood specimens can only be recorded in the LIS using the SCC lab module.
When the reports come back from a reference lab, the reporting date, reference lab number and result are
entered. A senior/charge technologist verifies the results in the LIS and the reference lab reports are
photocopied and then forwarded to the appropriate clinic or doctor.

a.        Log on: ‗lab‘

b.        Press 1-Order/Entry

c.        Press F3, scan barcode on reference lab requisition, Press F12

d.        Check that it is the correct patient

e.        Press F8 for result field

f.        Check that referred out tests have been ordered for all results coming back from the reference lab.
          If not, follow steps (i. to vi) for free text referred-out tests or proceed to step p for specific (non
          free text) refer-out tests:

                    i.         F1
                    ii.        Move cursor to next blank test field
                    iii.       On the Microbiology Order/Entry screen press W for Serology tests
                    iv.        Press S for Referred-out Test (8REF for MSH and 9MIS for UHN patients)
                    v.         Press F8 for result field
                    vi.        Press 1 To PHL

          Skip steps (vii – xi) if send-out tests results are complete (go to step g):

                    vii.       Press 4 @MOHR on the 8REF or 9MIS result field
                    viii.      Press \ to see the canned message ―This sample has been referred to the Public
                               Health Lab (PHL) 81 Resources Rd., Etobicoke, Ont., M9P 3T1 for the tests listed
                               below…‖ will appear
                    ix.        Type in the referred-out test(s)
                    x.         Press F12, F12
                    xi.        Yes to confirm



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 111
                                                                          Policy # MI/SER/15/04/v01                        Page 2 of 3
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Appendix IV - Entering Serology
                                                                                         Refer-Out Results

Continue from steps (g-o) below if referred-out tests are already ordered and results are ready to be
entered:

g.        Press F8 to enter refer-out results

h.        Select the appropriate test and enter the result from the keypad. If no keypad appears, type in "TO
          PHL" or ‗TO HSC‖ as appropriate in the result field (only 9 characters allowed)

i.        Delete the canned message @MOHR ―This sample has been referred to…‖ but leave the referred
          out tests names

j.        Free text the results by opening the text window by pressing \ in the result field and type in the
          results after each referred out test name

k.        After the last result, enter the reporting date, reference lab number either by free text or by
          pressing F5-Canned message and select 31 "@9rep".

l.        Space, enter date that PHL reported the result. Arrow up, wand PHL bar code or enter PHL lab
          number. F12, F12, Confirm modification? "Y".

m.        Record the Clinic, Dr., MRN, and order number on the reference lab report if not already
          recorded.

n. Give reports to senior/charge technologist to verify.

o.        Forward reports to Report Controllers for photocopying and mailing.


Alternate Method for non free-text specific refer-out tests, continued from steps (a-f) above:

p.        Specific (non free text) refer-out tests as can be ordered as follows:
                  -F1
                  -Move cursor to next blank test area
                  -^A-turn off keypad
                  -enter 9, F2, F12 to get a list of all send-out tests
                  -highlight the test to be ordered, ENTER
                  -repeat until all tests have been ordered
                  -F12, Confirm modification: Yes

q.        F8 to enter refer-out results


                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 112
                                                                          Policy # MI/SER/15/04/v01                        Page 3 of 3
                      Microbiology Department
Policy & Procedure Manual
Section: Serology Manual                                                  Subject Title: Appendix IV - Entering Serology
                                                                                         Refer-Out Results

r.        Highlight the appropriate test and enter the result from the keypad. If no keypad appears, type in
          "SEE BELOW".

s.        After the last result is entered, highlight the last result: \ - free text, F5-Canned message: select 7.
          "@9rep".

t.        Space, enter date that PHL reported the result. Arrow up, wand PHL bar code or enter PHL lab
          number. F12, F12, Confirm modification? "Y".

u.        Record the Clinic, Dr., MRN, and order number on the reference lab report if not already
          recorded.

v.        Give reports to senior/charge technologist to verify.

w.        Forward reports to Report Controllers for photocopying and mailing.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 113
                                                                               Policy # MI\SER\15\05\v04                          Page 1 of 2
                       Microbiology Department
 Policy & Procedure Manual
 Section: Serology Manual                                                      Subject Title: Appendix V - Autoverification
                                                                                              Process
 Issued by: LABORATORY MANAGER                                                 Original Date: June 25, 2002
 Approved by: Laboratory Director                                              Revision Date:
                                                                               Annual Review Date: January 10, 2011

                                 APPENDIX V - AUTOVERIFICATION PROCESS

I.        Introduction

          To document the autoverification process.

          Autoverification is the process by which the computer performs the initial verification of test
          results. Any value, that falls outside of the defined criteria, must be assessed by a technologist
          before release of results.

II.       Procedure

      1. The laboratory has a policy signed by the laboratory director approving the autoverification
         procedure.

      2. The results of autoverification is thoroughly tested, appropriately documented and signed by
         the section head/designee before implementation.

      3. If changes are made to the autoverification rules initially chosen and documented, the process
         is reverified as to its accuracy.

      4. The autoverification process is checked yearly.

AUTOVERIFICATION FOR THE AXSYM INSTRUMENT
POLICY:

The Axsym instrument performs the following tests for the Serology department:
1. Hepatitis B Surface Antigen
2. Hepatitis B Surface Antibody
3. Hepatitis B Core Antibody
4. Hepatitis A IgM Antibody
5. Hepatitis B Core IgM Antibody
6. Hepatitis Be Antigen
7. Hepatitis Be Antibody
8. Hepatitis C Antibody
9. Rubella Antibody
10. Cytomegalovirus IgG Antibody
11. Transplant Screen

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 114
                                                                               Policy # MI\SER\15\05\v04                   Page 2 of 2
                       Microbiology Department
 Policy & Procedure Manual
 Section: Serology Manual                                                      Subject Title: Appendix V -
                                                                                              Autoverification Process

Autoverification will occur as follows:

TEST NAME                     SOFT           RESULT                      TRANSLATION ACTION                                       REFLEX
                              TEST                                                                                                TESTS
                              CODE
Hep B Surface Ag              8HAG           NEGATIVE                    Negative                     Autoposted
                                                                                                      Autoverified

Hep B Surface Ag              8HAG           REACTIVE                    Review?                      NOT Autoposted              8HBC
                                                                                                      NOT                         Hep B
                                                                                                      Autoverified                Core Ab

Hep B Surface Ab              8HAB           REACTIVE                    POSITIVE                     Autoposted
                                                                                                      Autoverified
Hep B Surface Ab              8HAB           NEGATIVE                    Negative                     Autoposted
                                                                                                      Autoverified
Hep B Surface Ab              8HAB           GZ-NEGATIVE                 Neg                          Not Autoposted
                                                                                                      Not Autoverified

Hep B Core Ab                 8HBC           REACTIVE                    REACTIVE                     Not Autoposted              8HBC2
                                                                                                      Not Autoverified
Hep B Core Ab                 8HBC           NEGATIVE                    Negative                     Autoposted
                                                                                                      Autoverified
Hep A IgM Ab                  8HAV           REACTIVE                    REACTIVE                     Not Autoposted
                                                                                                      Not Autoverified
Hep B Core IgM                8HBCM          REACTIVE                    POSITIVE                     Not Autoposted              8HBC2
Ab                                                                                                    Not Autoverified
Hep Be Antibody               8HBEB          NONREACTIVE NONREACTIVE                                  Autoposted
                                                                                                      Autoverified
Hep Be Antibody               8HBEB          REACTIVE                    REACTIVE                     Autoposted
                                                                                                      Autoverified
Hep Be Antigen                8HBEG          NONREACTIVE NONREACTIVE                                  Autoposted
                                                                                                      Autoverified
Hep Be Antigen                8HBEG          REACTIVE                    REACTIVE                     Autoposted
                                                                                                      Autoverified
Rubella                       8RUB           NEGATIVE                    Negative                     Autoposted
                                                                                                      Autoverified
Rubella                       8RUB           GRAYZONE                    Negative                     Autoposted
                                                                                                      Autoverified



                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 115
                                                                               Policy # MI\SER\15\06\v04                   Page 1 of 1
                      Microbiology Department
 Policy & Procedure Manual
 Section: Serology Manual                                                      Subject Title: Appendix VI - Shipment of
                                                                                              Samples to HSC
 Issued by: LABORATORY MANAGER                                                 Original Date: Dec 20,2003
 Approved by: Laboratory Director                                              Revision Date: May 20, 2005
                                                                               Annual Review Date: January 10, 2011

                                APPENDIX VI - SHIPMENT OF SAMPLES TO HSC


          1. Fill out a HSC requisition. (Molecular tests require a colour coded UHN/MSH refer out
             Requisition to HSC Molecular Microbiology, Lab no. and tech initials)
          2. Put specimen and requisition in biohazard bag, and then in a brown paper bag.
          3. Put ‗Hospital for Sick Children, Microbiology Receiving,3rd Floor Atrium, Rm3676‘
             sticker on brown bag.
          4. Put labeled brown bag into blue specimen container, and send to ‗TG specimen
             management‘.
          5. Enter in LIS as ‗specimen send to HSC‘ and finalize.
          6. Also enter in serology send out binder.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 116
                                                                             Policy # MI\SER\15\07\v04                   Page 1 of 1
                       Microbiology Department
 Policy & Procedure Manual
 Section: Serology Manual                                                    Subject Title: Appendix VII - Posting of
                                                                                            AxSYM results
 Issued by: LABORATORY MANAGER                                               Original Date: Dec 20,2003
 Approved by: Laboratory Director                                            Revision Date:
                                                                             Annual Review Date: January 10, 2011

                                 APPENDIX VII - POSTING OF AxSYM RESULTS

1. Go to ‗lab‘
1.  ‗8‘-‗Inerface‘.
2.  ‗I‘-‗Interface Menu‘.
3.  ‗9‘-‗AXSYM‘.
4.  ‗Open‘ will be highlighted, ‗Enter‘.
5.  ‗Choose today‟s day‘ and ‗Enter‘.
6.  ‗P‘-Posting:
    ‗How to Post‘: change to‘ By Order‟ using space bar.
    ‗starting From: enter LIS # to be posted‘.
    ‗F12‘.
7. Do you want to verify results with Posting (Y/N)?. Press ‗N‘.
8. Reflex window will appear, ‗F12‘.
9. Press ‗[‘ to post result.
    Verify result with posting? Press ‗N‘.
    Under ST column ‗O‘ will change to ‗P‘.
10. ‗F12‘ to go to the next reflex, repeat steps 10.
11. At the end , press ‗F1‟ to exit.

                                   Test to be posted:                   Report as:
HBsAg+                             HBC                                  HBsAg: Positive
HBC +                              HBC 2: .x HBsAg +

HBC+                               HBC2                                 HBC: Positive
HBC2 +

HCV +, S/CO >10.00   HCV 2                                              HCV : Positive
HCV 2 +, S/CO >10.00

HCV +,S/CO <10.00                  HCV2                                 Send to PHL
HCV 2                                                                   Not tested




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 117
                                               Policy # MI\SER\15\08\v04       Page 1 of 1
                       Microbiology Department
 Policy & Procedure Manual
 Section: Serology Manual                      Subject Title: Appendix VIII - Looking Up
                                               Previous Hepatitis Results in EPR
 Issued by: LABORATORY MANAGER                 Original Date: Dec 20,2003
 Approved by: Laboratory Director              Revision Date:
                                               Annual Review Date: January 10, 2011

            APPENDIX VIII - LOOKING UP PREVIOUS HEPATITIS RESULTS IN EPR

1. Log on ‗EPR‘.
2. Enter‘ user ID‘ & ‗Password‘.
     1.Click on ‗All UHN Patients‘.
     2.Enter ‗MRN # in ‗Patient ID‘, press ‗Enter‘.
     3.Click on ‗any visit‘, then click on ‗Goto Selected visit‘.
     4.Click on ‗Chart Review‘, ‗Continue?‘,Click on‘ (y) yes‘.
     5.Click on ‗Hepatitis Profile‘, click on ‗OK‘.
     6.All previous Hepatitis test results will be displayed, e.g. HBsAg, HBsAb, HBcAb IgM, HBeAg,
       HBeAb, HC Ab, and HAAb IgM.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 118
                                                                             Policy # MI\SER\15\09\ v04                    Page 1 of 1
                       Microbiology Department
 Policy & Procedure Manual
 Section: Serology Manual                                                    Subject Title: Appendix IX
                                                                                    Printing of Pending List
 Issued by: LABORATORY MANAGER                                               Original Date: Dec 20,2003
 Approved by: Laboratory Director                                            Revision Date:
                                                                             Annual Review Date: January 10, 2011

                                   APPENDIX IX - PRINTING OF PENDING LIST


                    1. Log on ‗Lab‘.
                    1. Enter ‗ID‘ &‘Password‘.
                    2. ‗3‘-Results
                    3. ‗View/Enter Results by Sel Tests‘.
                    4. Select tests by ‗Template‘,‘Enter‘.
                    5. Enter ‗8SERO‘ under ‗Template‘, ‗Status- pend +nonver‘.
                    6. From order( enter last month‘s lab #) to‘ leave it blank‘.‘F12‘.
                    7. ‗F9‘ to print. Choose either ‗TC2RVIR‘ or ‗TC3R MIC‘ printer to print.
                    8. Look up each record, and find out why the results are still pending.




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 119
                                                                             Policy # MI\SER\15\10\v01                   Page 1 of 1
                       Microbiology Department
 Policy & Procedure Manual
 Section: Serology Manual                                                    Subject Title: Appendix X - Entering
                                                                                            Referred Test Results
 Issued by: LABORATORY MANAGER                                               Original Date: March 09, 2009
 Approved by: Laboratory Director                                            Revision Date:
                                                                             Annual Review Date:

                           APPENDIX X - ENTERING REFERRED TEST RESULTS


See Serology Referred Out Test Results Entry MI_SER_15_10.xls




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 120
Record of Edited Revisions

Manual Section Name: Serology

                      Page Number / Item                                             Date of Revision                   Signature of
                                                                                                                         Approval
Annual Review                                                                      May 12, 2003                       Dr. T. Mazzulli
Molecular Testing - HBV DNA-Reporting Added Para.                                  October 10, 2003                   Dr. T. Mazzulli
AxSym System-Reporting Results HBsAg                                               October 28, 2003                   Dr. T. Mazzulli
AxSym - CMV IgG Antibody (x)                                                       October 28, 2003                   Dr. T. Mazzulli
AxSym – Quality Control External Controls                                          October 28, 2003                   Dr. T. Mazzulli
AxSym - Added Failed QC                                                            October 28, 2003                   Dr. T. Mazzulli
AxSym HBsAg Surface Antigen - Pt. Samples                                          January 22, 2004                   Dr. T. Mazzulli
Appendix V (f) Added Steps                                                         January 15, 2004                   Dr. T. Mazzulli
HCV-RNA PCR Reporting - Positives                                                  April 02, 2004                     Dr. T. Mazzulli
Annual Review                                                                      May 26, 2004                       Dr. T. Mazzulli
Page 6 – Interpretation of Results – OD changed for                                May 20, 2005                       Dr. T. Mazzulli
Positive and Interdeterminate.
Page 6, 15 – Run external control – include QC and                                 May 20, 2005                       Dr. T. Mazzulli
instrument problems
Page 8 – re-centrifuge cloudy, previously frozen,                                  May 20, 2005                       Dr. T. Mazzulli
reconstituted samples.
Page 8 - Refer to Abbott Operation Manual vol. 1&2 for                             May 20, 2005                       Dr. T. Mazzulli
specific maintenance procedures
Page 13, 14 – report to MOH added                                                  May 20, 2005                       Dr. T. Mazzulli
Page 13, 14 – centrifuge before repeating                                          May 20, 2005                       Dr. T. Mazzulli
Page 16, file log and external control out of range                                May 20, 2005                       Dr. T. Mazzulli
Page 48 – store VDRL samples for 3 months after testing                            May 20, 2005                       Dr. T. Mazzulli
Page 51 – 56 VZV Vidas revised                                                     May 20, 2005                       Dr. T. Mazzulli
Page 57 – WNV IgG not currently in use                                             May 20, 2005                       Dr. T. Mazzulli
Page 61 – WNV IgM not currently in use                                             May 20, 2005                       Dr. T. Mazzulli
Page 67 - Molecular Testing - Chlamydia Trachomatis &                              May 20, 2005                       Dr. T. Mazzulli
Neisseria gonorrhoeae
Page 67 – urine but not more than 60 mL added                                      May 20, 2005                       Dr. T. Mazzulli
Page 68 – specimen not lysed…added                                                 May 20, 2005                       Dr. T. Mazzulli
Page 68 – tubes may be blotted                                                     May 20, 2005                       Dr. T. Mazzulli
Page 70 – handling samples after lysing                                            May 20, 2005                       Dr. T. Mazzulli
Page 71,72 Procedure change                                                        May 20, 2005                       Dr. T. Mazzulli
Page 72, interpretation of result - change MOTA score                              May 20, 2005                       Dr. T. Mazzulli
Page 73, reporting – indeterminate                                                 May 20, 2005                       Dr. T. Mazzulli
Page 76 – do not vortex working master mix                                         May 20, 2005                       Dr. T. Mazzulli
Page 76 – specimen dilution protocol                                               May 20, 2005                       Dr. T. Mazzulli
Page 77 – 80 procedure and reporting changed                                       May 20, 2005                       Dr. T. Mazzulli
Page 81 – QC procedure changed                                                     May 20, 2005                       Dr. T. Mazzulli
Page 93 – procedure changed                                                        May 20, 2005                       Dr. T. Mazzulli
Page 86 – 88, instrument instructions                                              May 20, 2005                       Dr. T. Mazzulli

                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 121
                      Page Number / Item                                             Date of Revision                   Signature of
                                                                                                                         Approval
Page 90 – WNV RT-PCR                                                               May 20, 2005                       Dr. T. Mazzulli
Page 92 – lysis buffer                                                             May 20, 2005                       Dr. T. Mazzulli
Page 93 – General Precaution section revised                                       May 20, 2005                       Dr. T. Mazzulli
Page 94-100 – procedure revised                                                    May 20, 2005                       Dr. T. Mazzulli
Page 101 – schedule revised                                                        May 20, 2005                       Dr. T. Mazzulli
Page 105 – shipping to St. Joseph‘s                                                May 20, 2005                       Dr. T. Mazzulli
Page 121 – shipping to HSC                                                         May 20, 2005                       Dr. T. Mazzulli
Annual Review                                                                      May 20, 2005                       Dr. T. Mazzulli
Page 128 – Updated decontamination procedure                                       June 2, 2005                       Dr. T. Mazzulli
Page 6 – C. difficile toxin indeterminate range changed                            December 29, 2005                  Dr. T. Mazzulli
Page 13 – HBc IgM reporting guide changed                                          December 29, 2005                  Dr. T. Mazzulli
Page 15 – MV, Rubella reporting                                                    May 29, 2006                       Dr. T. Mazzulli
Annual Review                                                                      May 29, 2006                       Dr. T. Mazzulli
Added links to TGLN Procedures                                                     March 21, 2007                     Dr. T. Mazzulli
Annual Review                                                                      June 8, 2007                       Dr. T. Mazzulli
Added Labour and Delivery STAT HBsAg and HIV                                       March 19, 2008                     Dr. T. Mazzulli
Instruction
Added Immuncor Capture-CMV Assay                                                   July 14, 2008                      Dr. T. Mazzulli
Revised Document numbers                                                           July 14, 2008                      Dr. T. Mazzulli
Added Ortho ELISA Assays for HBcore, HCV,                                          July 14, 2008                      Dr. T. Mazzulli
HTLVI/II for Donor Testing
Annual Review                                                                      July 14, 2008                      Dr. T. Mazzulli
Report positive C. difficile toxin as an ISOLATE in the                            September 16, 2008                 Dr. T. Mazzulli
LIS
Serology Referred Out Test Results Entry Phrases added                             March 09, 2009                     Dr. T. Mazzulli
Replaced HBsAg, HIV 1/2 HBcAb, HCV Ab, HTLV I/II                                   November 18, 2009                  Dr. T. Mazzulli
Ab donor serology sections
Added Aspergillus Galactomannan Antigen Detection                                  November 18, 2009                  Dr. T. Mazzulli
Assay
Annual Review                                                                      November 18, 2009                  Dr. T. Mazzulli
Architect System added                                                             May 17, 2010                       Dr. T. Mazzulli
C. difficile Toxin EIA edited with Premier kit                                     May 17, 2010                       Dr. T. Mazzulli
Annual Review                                                                      January 10, 2011                   Dr. T. Mazzulli
Remove C. difficile toxin EIA                                                      January 10, 2011                   Dr. T. Mazzulli
Syphillis screen added to Architect System                                         January 31, 2011                   Dr. T. Mazzulli
Modified RPR for Cadaver donors                                                    January 31, 2011                   Dr. T. Mazzulli
Updated Axsym procedure                                                            January 31, 2011                   Dr. T. Mazzulli
Updated VZV procedure                                                              January 31, 2011                   Dr. T. Mazzulli
Updated Appendix II                                                                January 31, 2011                   Dr. T. Mazzulli




                                                          PROCEDURE MANUAL
                     UNIVERSITY HEALTH NETWORK / MOUNT SINAI HOSPITAL MICROBIOLOGY DEPARTMENT
   NOTE: This is a CONTROLLED document for internal use only. Any documents appearing in paper form are not controlled and should be checked
                                        against the document (titled as above) on the server prior to use.
C:\Docstoc\Working\pdf\3b93f20a-f286-4299-86d0-6f9f11e50df1.doc
                                                                  Page 122

				
DOCUMENT INFO