IQ/OQ/PQ Validation Document

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					IQ/OQ/PQ Validation Document
1.0   Purpose
      1.1   The purpose of this document is to assist customers in the validation of the Pall
            eBDS. Pall Medical has validated the Pall eBDS Oxygen Analyzer and Sample Set in
            accordance with FDA regulations. Installation Qualification (IQ), Operations
            Qualification (OQ), and Performance Qualification (PQ) are recommended during
            initial implementation of the Pall eBDS. The Pall eBDS Oxygen Analyzer is calibrated
            annually. Upon installation of a newly calibrated analyzer, it is recommended that
            analyzer performance be verified by conducting the IQ and OQ portions of the
            document.
2.0   Equipment and Materials
      2.1   Leukocyte-reduced Platelet Product or Surrogate
      2.2   Pall eBDS Sample Set
      2.3   Pall eBDS Oxygen Analyzer
      2.4   Metrologic® Barcode Scanner
      2.5   Oxygen Sampling Stand
      2.6   Computer
      2.7   Printer
      2.8   Data Capture Software
      2.9   Tubing Stripper
      2.10 Sterile Connection Device (SCD) and Wafers
      2.11 Clamp or Hemostat
      2.12 Tube Sealer
      2.13 Helmer Platelet Incubator Capable of Maintaining 35 ± 1 °C
      2.14 Helmer Flatbed Platelet Agitator
      2.15 Platelet Storage Area or Incubator Capable of Maintaining 22 ± 2 °C
3.0   Scope
      3.1   This document applies to staff that have completed training on the Pall eBDS.
4.0   Data Capture and Printing
      4.1   The Pall eBDS Oxygen Analyzer is configured for data logging. The analyzer is
            compatible with hardware that has a serial port (RS232). Data capture is
            accomplished by directly interfacing the analyzer with a computer system installed
            with data capture software. Please refer to the Pall eBDS Oxygen Analyzer User’s
            Guide for the information logged by analyzer. A Pall Medical Product Application
            Specialist can assist during set up to configure the data capture system.
5.0   Incubator/Agitator Set Up
      5.1   The Pall eBDS requires incubation and agitation of the sample pouches at 35 ± 1 °C.
            Pall Medical provides Helmer Platelet Incubators and Agitators to Pall eBDS
            customers as part of a turnkey system. Installation and operation should be
            conducted as per the operation manuals supplied by the vendor. Pall Medical
            Product Application Specialists are available to assist with instrument set up if
            needed.
6.0   References
      6.1   Pall eBDS Oxygen Analyzer User’s Guide
      6.2   Pall eBDS Sample Set Package Insert
      6.3   Helmer Platelet Incubator User’s Manual

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      6.4   Helmer Flatbed Platelet Agitator User’s Manual
      6.5   Metrologic Single Line Installation and User’s Guide
      6.6 Metrologic Single Line Configuration Guide
7.0   Set Up Checklist
A Pall Medical Product Application Specialist will complete the following:
______ Unpack and confirm Pall eBDS Oxygen Analyzer inventory against packing list
______ Change fuses from 0.63AT to 1.6AT
______ Adjust voltage setting to 115VAC
______ Connect sample hose to analyzer
______ Connect power cord to the analyzer and then to power outlet
______ Connect barcode scanner to analyzer
______ Connect analyzer to computer
______ Turn on the Pall eBDS Oxygen Analyzer
______ Configure barcode scanner
______ Configure Pall eBDS Oxygen Analyzer barcode settings
______ Adjust Pall eBDS Oxygen Analyzer date and time settings
______ Install data capture software
______ Apply asset tags (if applicable)
______ Verify all serial numbers match (analyzer, box, and calibration certificate)


Pall eBDS Oxygen Analyzer Serial Number: ________________
Pall eBDS Oxygen Analyzer Hardware Version: _____________
Pall eBDS Oxygen Analyzer Software Version: _____________
Facility:_______________________________________________


I certify that the Pall eBDS Oxygen Analyzer has been set up and configured as indicated in the
Pall eBDS Oxygen Analyzer user guide (part no. ______________ issue date ___________).


Product Application Specialist or designee: ___________________________________________
Date _________________________________________________________________________




Last updated 11/22/2006                                                                       2
8.0     Installation Qualification
        The installation qualification verifies that the Pall eBDS Oxygen Analyzer has been
        appropriately configured and is ready to conduct measurements after the warm up period.
        The section includes instrument start up, barcode scanning and data capture. The Pall
        eBDS Oxygen Analyzer can be configured to scan for any or all of the following: User ID,
        Lot Number, Product Code, and Donation ID. Indicate “Not Applicable” in appropriate
        sections for barcode options not utilized. For changes to barcode scanning options, refer to
        the Pall eBDS Oxygen Analyzer User’s Guide. After completion of the installation
        qualification, generate a printed report of the data.


        8.1.   Start Up
 Step          Procedure                     Expected Result                    Meets        Initials
                                                                                Expected     and
                                                                                Result Y/N   Date
 8.1.1         Turn on the Pall eBDS         Instrument conducts a self-
               Oxygen Analyzer.              test. Software version and
                                             self-test are briefly displayed.
                                             Warm up screen is displayed
                                             and includes 30-minute
                                             warm up.
 8.1.2         Verify date and time.         “Date” and “Time” are
               To adjust date and time,      displayed during warm up
               refer to section 5 “Setup     cycle.
               Menu” in the User’s Guide
 8.1.3         Ready to measure sample       Baseline reading is
               pouch is displayed after 30   automatically conducted at
               minute warm up.               the end of warm up cycle.
                                             “Ready to measure sample
                                             pouch” is displayed to
                                             indicate the system is ready.
                                             If barcode scanning options
                                             are utilized the screen will
                                             display a prompt to scan the
                                             indicated barcode.
VALIDATION AND TECHNICAL SUPPORT
        8.2    Barcode Scanning and Data Capture
 Step          Procedure                     Expected Result                    Meets        Initials
                                                                                Expected     and
                                                                                Result Y/N   Date
 8.2.1         Turn on data capture          The data capture software is
               software.                     ready to accept data.
 8.2.2         In the main menu on the       Enter passcode screen is
               Pall eBDS Oxygen              displayed.
               Analyzer, press the right
               hand soft key.
 8.2.3         Using the fixed keys, enter   Access is granted to the set
               the User Supervisor           up and diagnostic menus.
               passcode and press the        (Indicated by the appearance
               green “Enter” button.         of set up and diagnostic
                                             icons.)



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 8.2.4    Press the center soft key    Setup 1 screen is displayed.
          below the Setup Menu
          icon.
 8.2.5    Press the right hand soft    Setup 2 screen is displayed.
          key below the Next Screen
          icon.
 8.2.6    Verify PC Handshake is       Setup 2 screen displays PC
          set to "Yes".                Handshake: Yes
 8.2.7    Press the right hand soft    Setup 3 screen is displayed.
          key below the Next Screen
          icon.
 8.2.8    Scan a User ID barcode.      The scanned barcode
                                       appears on Setup 3 screen.
 8.2.9    Press the right hand soft    Setup 4 screen is displayed.
          key below the Next Screen
          icon.
 8.2.10   Scan a Lot Number            The scanned barcode
          barcode.                     number appears on Setup 4
                                       screen.
 8.2.11   Press the right hand soft    Setup 5 screen is displayed.
          key below the Next Screen
          icon.
 8.2.12   Scan a Product Code          The scanned barcode
          barcode.                     number appears on Setup 5
                                       screen.
 8.2.13   Press the right hand soft    Setup 6 screen is displayed.
          key below the Next Screen
          icon.
 8.2.14   Scan a Donation ID           The scanned barcode
          barcode.                     number appears on Setup 6
                                       screen.
 8.2.15   Go to any barcode-           The appropriate Setup
          scanning menu that has       screen is displayed.
          been previously
          programmed for ISBT128
          Codabar, or USS Codabar.
          Example-Setup 4 screen,
          which is configured to
          accept ISBT128.
 8.2.16   Scan a barcode               The Screen displays "Invalid
          incompatible with the        Input".
          indicated (configured)
          symbology.
          Example-scan a Codabar
          barcode in a field that is
          configured for ISBT128.
 8.2.17   Press the soft key below     The selected Setup menu is
          the checkmark to             displayed.
          recognize the alarm.



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 8.2.18    Press the left hand soft      A baseline is conducted and
           key twice to return to the    the main menu is is
           main menu.                    displayed.
 8.2.19    Scan in barcodes as           "Ready to measure sample
           prompted on main menu         pouch" and the scanned
           screen.                       barcodes are displayed.



 8.2.20    Leave probe in holder and     A green screen with "Pass",
           press green "Enter" button.   date/time, and Donation ID is
           Follow prompts to             displayed.
           complete sample
           measurement.
 8.2.21    Press soft key below          The measurement is
           check mark to return to       accepted and the main menu
           main menu.                    is displayed.
 8.2.22    Verify data has transferred   The results of the baseline
           to the data capture           and sample measurement
           software. Refer to User       will be displayed on
           Guide section 7 for           computer screen.
           description and example of
           data log.
                                         If barcode information was
                                         scanned, that information will
                                         be displayed. Otherwise the
                                         incremental donation ID will
                                         be displayed and the
                                         remaining fields will be blank.


Comments: ___________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________


Pall eBDS Oxygen Analyzer Serial Number: __________________________________________
Pall eBDS Oxygen Analyzer Hardware Version:_______________________________________
Pall eBDS Oxygen Analyzer Software Version: ________________________________________
Metrologic® Barcode Scanner Make: _______________________________________________
Metrologic Barcode Scanner Model Number: _________________________________________
The installation qualification testing meets expected criteria ______ Yes ______ No


Signature: ______________________________________________________ Date:__________




Last updated 11/22/2006                                                               5
9.0     Operations Qualification
The operations qualification verifies that the Pall eBDS Oxygen Analyzer functions as indicated
during sample pouch measurements. The PASS/FAIL display and alarms will be tested. The
operations qualification protocol below uses data capture software to collect the data. If manual
data capture is desired, refer to the Pall eBDS Oxygen Analyzer User’s Guide section 5.
Note: To expedite the Operations Qualification Protocol, barcode scanning may be disabled. The
steps below reflect disabled barcode scanning. If scanning barcodes, follow the prompts
displayed on the analyzer and scan appropriate barcodes until the “Ready to measure sample
pouch” is displayed. After completion of the Operations Qualification, generate a printed report of
the data.
        9.1   Measurement of Sample Pouch to Produce Pass Reading
 Step         Procedure                     Expected Result               Meets         Initials
                                                                          Expected      and
                                                                          Result Y/N    Date
 9.1.1        Verify analyzer is at main    Screen displays "Pall eBDS
              menu and "Ready to            ready to measure sample
              measure sample pouch" is      pouch".
              displayed.
 9.1.2        Leave probe in holder and     Screen displays "WAIT,
              press green "Enter" key to    measuring sample pouch".
              conduct sample
              measurement.
 9.1.3        Press any key to make an      Pressing keys has no effect
              entry while pump is           on the sample measurement.
              operating.
 9.1.4        Press the green "Enter"       Screen displays "WAIT,
              key to confirm baseline.      measuring baseline".
 9.1.5        Press any key to make an      Pressing keys has no effect
              entry while pump is           on baseline reading.
              operating.
 9.1.6        Verify that "Pass" is         A green screen with "Pass",
              displayed on screen.          date/time, and donation ID
                                            are displayed.
 9.1.7        Press soft key below          The measurement is
              checkmark to return to        accepted and the main menu
              main menu.                    is displayed.
 9.1.8        Verify data is displayed on   The sample pouch and
              data capture screen.          baseline results are
                                            displayed on the data
                                            capture software screen.
 9.1.9        Conduct an additional two     Two additional Pass
              oxygen measurements           readings were produced and
              and verify data capture.      displayed on the data
                                            capture screen.


Comments: ___________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________




Last updated 11/22/2006                                                                            6
9.2 Measurement of Sample Pouch to Produce FAIL Reading
 Step     Procedure                     Expected Result                 Meets        Initials
                                                                        Expected     and
                                                                        Result Y/N   Date
 9.2.1    Verify analyzer is at    Screen displays "Pall eBDS
          main menu and "Ready     ready to measure sample
          to measure sample pouch" pouch".
          is displayed.
 9.2.2    Obtain a prepared low         Screen displays "WAIT,
          oxygen content sample.        measuring sample pouch".

          Insert probe of the Oxygen
          Analyzer into the sample.

          Press the green "Enter"
          key to conduct sample
          measurement.
 9.2.3    Follow prompt and remove      Screen displays "WAIT,
          probe from sample.            measuring baseline".

          Press the green "Enter"
          key to confirm baseline.
 9.2.4    Verify that a flashing        Audible alarm and a red
          "FAIL" is displayed on        flashing screen with "FAIL",
          screen and an audible         date/time, and donation ID is
          alarm is present.             displayed.
 9.2.5    Press soft key below          After a "FAIL" reading, the
          checkmark to return to        flow system is flushed out.
          main menu.                    The screen displays "Wait"
                                        and a count down is shown.
                                        The main menu is displayed
                                        after system flush.
 9.2.6    Verify data is displayed on   The sample pouch and
          data capture screen.          baseline results are
                                        displayed on the data
                                        capture software screen.
 9.2.7    Conduct two additional        Two additional "FAIL"
          "FAIL" measurements and       readings were produced and
          verify data capture.          displayed on the data
                                        capture screen.


Comments: ___________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________




Last updated 11/22/2006                                                                         7
9.3 Alarms Testing
 Step     Procedure                     Expected Result                Meets        Initials
                                                                       Expected     and
                                                                       Result Y/N   Date
 9.3.1    Verify analyzer is at main    Screen displays "Pall eBDS
          menu and "Ready to            ready to measure sample
          measure sample pouch" is      pouch".
          displayed.
 9.3.2    Loosen the finger screw       An Alarm message is
          connecting the tubing to      displayed. "Sample hose
          the analyzer.                 error. Check filter, probe,
                                        hose and connections".
          Press the green "Enter"
          key to conduct sample
          measurement.
 9.3.3    Tighten the finger screw      A baseline measurement is
          connecting the tubing to      conducted and the screen
          the analyzer.                 displays "Ready to measure
                                        a sample pouch".
 9.3.4    Remove probe from holder      An Alarm message is
          and block the end of the      displayed. "Filter blocked.
          probe.                        Change probe and filter."
 9.3.5    Release probe blockage.       A baseline measurement is
                                        conducted and screen
          Press the soft key below      displays "Ready to measure
          the checkmark on the          a sample pouch."
          screen.
 9.3.6    Obtain small plastic bag.     An Alert is displayed.
                                        "Unable to confirm baseline.
          Inhale, then exhale           Replace probe and filter and
          completely into bag OR        press "Enter" to confirm
          obtain a low oxygen           baseline."
          concentration sample.

          Insert probe into sample or
          bag and close seal of bag
          around probe.

          Press the soft key below
          the checkmark on the main
          menu screen.
 9.3.7    Leave probe in bag and        An Alert is displayed.
          press the green "Enter"       "Unable to confirm baseline.
          key.                          To reset baseline, press the
                                        (20, 9) button."
 9.3.8    Remove probe from bag.        Screen displays "Wait". The
          Press soft key below          baseline is reset and the
          20.9% O2.                     main menu is displayed.


Comments: ___________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________
Last updated 11/22/2006                                                                        8
9.4 Hand Shaking
 Step     Procedure                    Expected Result              Meets        Initials
                                                                    Expected     and
                                                                    Result Y/N   Date
 9.4.1    Verify analyzer is at main   Screen displays "Pall eBDS
          menu and "Ready to           ready to measure sample
          measure sample pouch" is     pouch".
          displayed.
 9.4.2    Detach the cable leading     An Alert is displayed. "PC
          to the computer from the     not connected."
          back of the Pall eBDS
          Oxygen Analyzer.
 9.4.3    Reconnect cable leading      Screen displays "Pall eBDS
          to the computer to the       ready to measure sample
          back of the Pall eBDS        pouch."
          Oxygen Analyzer. Press
          the soft key below
          checkmark to return to
          main menu.


Comments: ___________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________




Last updated 11/22/2006                                                                     9
9.5 Leak Test
 Step     Procedure                      Expected Result                Meets        Initials
                                                                        Expected     and
                                                                        Result Y/N   Date
 9.5.1    Verify analyzer is at main     Screen displays "Pall eBDS
          menu and "Ready to             ready to measure sample
          measure sample pouch" is       pouch".
          displayed.
 9.5.2    Press the soft key below       Screen displays "BLOCK
          the center (leak test) icon.   PROBE. Press enter to start
                                         leak test of probe assy."
 9.5.3    Press the end of the probe     Screen displays "Wait,
          securely against the           testing probe assy."
          rubber disc on the Oxygen
          Analyzer and press the         "No leak detected."
          green enter key.
 9.5.4    Press the soft key below       The main menu is displayed.
          the checkmark to return to
          the main menu.
 9.5.5    Press the soft key below       Screen displays "BLOCK
          the center (leak test) icon.   PROBE. Press enter to start
                                         leak test of probe assy."
 9.5.6    Leave the probe in the         An alert is displayed.
          probe holder and press the     "Unable to confirm probe
          green enter key.               assy test. Replace probe and
                                         filter. BLOCK PROBE. Press
                                         enter to test probe assy."
 9.5.7    Leave the probe in the         An alert is displayed.
          probe holder and press the     "Unable to confirm probe
          green enter key.               assy test. Replace complete
                                         probe assy. BLOCK PROBE.
                                         Press enter to test probe
                                         assy."
 9.5.8    Place the end of the probe     Screen displays "No leak
          securely against the           detected."
          rubber disc on the Oxygen
          Analyzer and press the
          green enter key.
 9.5.9    Press the soft key below       A baseline measurement is
          the checkmark to return to     conducted then the main
          the main menu.                 menu is displayed.


Comments: ___________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________


The operation qualification testing meets expected criteria ______Yes ______No
Signature:________________________________________________ Date: ________________




Last updated 11/22/2006                                                                    10
10.0 Performance Qualification
The performance qualification verifies that the Pall eBDS will function as intended. The process
includes sample collection using the Pall eBDS Sample Set and oxygen analysis using the Pall
eBDS Oxygen Analyzer. Please refer to the Pall eBDS Sample Set package insert for sample
collection and the Pall eBDS Oxygen Analyzer User’s Guide for oxygen measurement
instructions. Customers who wish to conduct the performance qualification using product that has
been inoculated with bacteria to produce a positive control should refer to Addendum A through C
for suggested guidelines.

 Step      Procedure                       Expected Result                  Meets        Initials
                                                                            Expected     and
                                                                            Result Y/N   Date
 10.1      Collect samples from            Sample pouches are
           leukocyte reduced               prepared for the verification
           platelets using the Pall        process.
           eBDS Sample Set as per
           instructions for use.

           Sample size for
           performance qualification
           is at the discretion of the
           Blood Establishment.

           Note: Thirty samples
           provide a minimum
           confidence level of 95%
           and reliability level of 90%.
 10.2      Measure the oxygen level        All samples measured will
           in the headspace of the         provide "Pass" or "Fail"
           pourch as per instructions      values.
           for use.
                                           Three sample pouches had
           Include within the product      two measurements per
           qualification conducting        sample pouch conducted.
           two measurements per
           sample pouch on a total of
           three sample pouches.
 10.3      Access data from data           A printed report is generated.
           capture software and
           generate a printed report
           of the performance
           qualification data.


Comments:
_____________________________________________________________________________
_____________________________________________________________________________
_____________________________________________________________________________


The performance qualification testing meets expected criteria ______Yes ______No
Signature:________________________________________________ Date: ________________



Last updated 11/22/2006                                                                        11
Addendum A

Suggested Guidelines For Conducting Positive Control Testing for the eBDS-SLS
Inoculation:

Purpose
To provide Pall Medical customers guidelines to assist with product qualification of the Pall eBDS
using inoculated leukoreduced platelet product used as a positive control.

Summary
Pall Medical has developed several options for customers to conduct positive control testing. See
Addendum A through C. This document outlines the steps for a customer to send a leukoreduced
platelet concentrate (PC) to Pall’s Scientific and Laboratory Services (SLS). SLS will inoculate the
PC with a low level inoculum (target 1-15 CFU/mL) and return the product to the customer. Upon
receipt, the customer will collect samples into the Pall eBDS Sample Set and measure the
oxygen concentration using the Pall eBDS Oxygen Analyzer as per instructions for use.

Scope
The document applies to staff that have completed training on the Pall eBDS.

Equipment and Materials
 Leukoreduced platelet product
 Controlled environment shipping containers
 eBDS equipment (see section 2 of IQ/OQ/PQ document)
 Serratia marcescens ATCC# 43862
 Staphlyococcus aureus ATCC #27217

Procedure
1.   Contact SLS to schedule a date for positive control testing, provide at least one week
     advance notice. Provide the following information: Facility Name, Address, Contact Name,
     Phone Number, and Email Address.
     Contact Information:
     Person: Lois Hayon
     Telephone: 516.801.9243
     Email: lois_hayon@pall.com
     Hours: 8:00 am - 5:00 pm (ET)
     Days: Monday - Friday
2.   Ship the leukocyte reduced platelet concentrates (in appropriate controlled environment
     shipping containers) to:
     Pall Corporation
     Attn. Lois Hayon
     25 Harbor Park Dr.
     Port Washington, NY 11050
3.   Notify SLS with day of shipment and tracking number.
4.   SLS will inoculate the PC with 1-15 CFU/mL and collect a sample to verify colony count.
5.   The platelets will be labeled with the bacteria type, date and time of inoculation.
6.   The inoculated PC will be shipped overnight to the customer.
7.   Upon receipt, place inoculated PC on platelet agitator at 22 ± 2 °C for approximately one
     hour.
8.   At least twenty-four hours after the indicated inoculation time, gently mix the unit and strip
     the line on the platelet bag. Ensure the tubing is filled with a well-mixed representative
     sample.
9.   Collect samples into the Pall eBDS Sample Set as per instructions for use.
10. Incubate pouches at 35 ± 1 °C as indicated in the Pall eBDS Sample Set instructions for
     use.
11. Measure oxygen concentration in the headspace of the pouch as per Pall eBDS Oxygen
     Analyzer User’s Guide.

Last updated 11/22/2006                                                                         12
Addendum B

Suggested Guidelines For Conducting Positive Control Testing for the eBDS-Customer
Inoculation:

Purpose
To provide Pall Medical customers guidelines to assist with product qualification of the Pall eBDS
using inoculated leukoreduced platelet product used as a positive control.

Summary
Pall Medical has developed several options for customers to conduct positive control testing. See
Addendum A through C. This document outlines the steps where Pall’s Scientific and Laboratory
Services (SLS) provide customers with vials of bacteria in a phosphate buffered suspension. The
customer determines the volume to inject into the leukoreduced platelet concentrate. Inoculates
the PC and then incubates at 22 ± 2 °C for twenty-four hours. The customer then collects
samples into the Pall eBDS Sample Set and measures the oxygen concentration using the eBDS
Oxygen Analyzer as per instructions for use.

Scope
The document applies to staff that have completed training on the Pall eBDS.

Equipment and Materials
 Leukocyte reduced platelet concentrate
 Electronic scale
 Site couplers
 Alcohol swabs
 1-3 cc syringe
 Large bore needle (16-18 gauge)
 eBDS equipment (see section 2 of IQ/OQ/PQ document)
 Serratia marcescens ATCC# 43862 in PBS
 Staphlyococcus aureus ATCC #27217 in PBS
 Biohazardous waste containers
 Sharps disposal containers
 Personal protective equipment

Procedure
1.   Contact SLS to schedule a date for positive control testing, provide at least one week
     advance notice. Provide the following information: Facility Name, Address, Contact Name,
     Phone Number, Email Address, Number of Tests, Type of Product, and Type of Bacteria.
     Contact Information:
     Person: Lois Hayon
     Telephone: 516.801.9243
     Email: lois_hayon@pall.com
     Hours: 8:00 am - 5:00 pm (ET)
     Days: Monday - Friday
2.   SLS will ship, on ice, two vials of bacteria in Phosphate Buffered Solution (PBS). The vials
     will be labeled with the following information: date of preparation, type of bacteria, and
     concentration.
     Note: The vials must be used within 24 hours.
3.   Upon receipt, allow the vials to come to room temperature (approximately 15 minutes ).
4.   Gently vortex to create a homogenous suspension of inoculum solution.
5.   Weigh leukocyte reduced platelet concentrate (PC).
6.   Determine volume of PC.
7.   Use the following calculation to determine the volume of inoculum to add to the platelet
     product.



Last updated 11/22/2006                                                                       13
      Target inoculum is 1-15 CFU/mL

      Inoculum Volume (mL) = Platelet Volume (mL) x Target Inoculum (CFU/mL)
                                               CFU/mL in Inoculation Tube
      Example:
      Inoculum Volume = 300 mL x 10 CFU/mL = 2 mL
                                        3
                              1.50 x 10 CFU/mL
8.    Using aseptic technique, place the site coupler into port of platelet bag.
9.    Clean injection site with alcohol and allow the site to air dry.
10.   Gently mix the designated vial and, using a syringe and needle, withdraw the calculated
      volume.
11.   Inject the volume into the platelet bag via site coupler.
12.   Flush the syringe with platelet concentrate several times to ensure all of the volume is
      dispensed.
13.   Place the inoculated platelet concentrate on a platelet agitator at 22 ± 2 °C for twenty-four
      hours.
14.   After 24 hours incubation, remove the inoculated platelet concentrate from the agitator.
15.   Gently mix the unit and strip the line on the platelet bag. Ensure the tubing is filled with a
      well mixed representative sample.
16.   Collect samples into the Pall eBDS Sample Set as per instructions for use.
17.   Incubate pouches at 35 ± 1 °C as indicated in the Pall eBDS Sample Set instructions for
      use.
18.   Measure oxygen concentration in the headspace of the pouch as per Pall eBDS Oxygen
      Analyzer User’s Guide.




Last updated 11/22/2006                                                                          14
Addendum C

Suggested Guidelines For Inoculating Leukoreduced Platelet Product-Unassisted Testing:

Purpose
To provide Pall Medical customers guidelines to assist with product qualification of the Pall eBDS
using inoculated leukoreduced platelet product used as a positive control.

Summary
Pall Medical has developed several options for customers to conduct positive control testing. See
Addendum A through C. This document outlines steps for the Blood Establishment to conduct
testing without the assistance of Pall Medical.

Scope
The steps in this document are to be conducted by a microbiologist and applies to staff that have
completed training on the Pall eBDS.

Equipment and Materials
 Trypticase soy broth (TSB)
 Sterile 0.9% saline
 Sterile transfer pipette
 Sterile tubes (10 mL)
 0.5 McFarland Standard
 Turbidity meter
 Trypticase soy agar with or without blood
 Leukoreduced platelet product
 Site couplers
 Alcohol swabs
 1-3 cc syringe
 Large bore needle (16-18 gauge)
 Agitator
 Serratia marcescens ATCC# 43862
 Biohazardous waste containers
 Sharps disposal containers
 Personal protective equipment

Procedure
1.   Obtain lyophilized bacteria from American Type Culture Collection (ATCC).
2.   Reconstitute lyophilized pellet as per instructions for use provided by ATCC.
3.   Transfer reconstituted pellet to a labeled 5 mL TSB tube.
4.   Incubate the TSB (bacterial culture) tube at 35 °C overnight (approximately 16-20 hours).
5.   Remove the bacterial culture from incubator and leave at room temperature for at least 30
     minutes to allow it to cool.
6.   Mix the bacterial culture well.
7.   Create a suspension of the bacterial culture in 0.9% sterile saline equivalent to a 0.5
     McFarland standard. This can be accomplished by adding one drop at a time to the 0.9%
     sterile saline and monitoring turbidity of the 0.9% sterile saline after each drop is added.
8.   Vortex the suspension before taking a reading from the turbidity meter. The 0.5 McFarland
     standard will provide approximately 1.50E + 08 CFU/mL bacteria (Tube #1).
9.   Create 1:10 serial dilutions from tube # 1 into 0.9% sterile saline. Example: 1 mL of
     suspension added to 9 mL of sterile 0.9% saline (see Table 1).




Last updated 11/22/2006                                                                       15
      Table 1
      Inoculation Tube #              Approximate Bacteria              Dilution
                                      Concentration
                                               8
      1                               1.50 x 10                         Equivalent to 0.5
                                                                        McFarland Std.
                                                7
      2                               1.50 x 10                         1:10
                                               6
      3                               1.50 x 10                         1:10
                                               5
      4                               1.50 x 10                         1:10
                                               4
      5                               1.50 x 10                         1:10
                                               3
      6                               1.50 x 10                         1:10

10.   Determine volume of leukoreduced platelet product.
11.   Use the following calculation to determine the volume of inoculum to add to the platelet
      product.
      Target inoculum is 1-15 CFU/mL

      Inoculum Volume (mL) = Platelet Volume (mL) x Target Inoculum (CFU/mL)
                                               CFU/mL in Inoculation Tube
      Example:
      Inoculum Volume = 300 mL x 10 CFU/mL = 2 mL
                                   3
                         1.50 x 10 CFU/mL (tube #6)
12.   Using aseptic technique, place the site coupler into port of platelet bag.
13.   Clean injection site with alcohol and allow the site to air dry.
14.   Mix designated tube and, using a syringe and needle, withdraw the calculated volume.
15.   Inject the volume into the platelet bag via site coupler.
16.   Flush the syringe with platelet concentrate several times to ensure all of the volume is
      dispensed.
17.   Place the inoculated platelet concentrate on a platelet agitator at 22 ± 2 °C for twenty four
      hours.
18.   After 24 hours incubation, remove the inoculated platelet concentrate from the agitator.
19.   Gently mix the unit and strip the line on the platelet bag. Ensure the tubing is filled with a
      well-mixed representative sample.
20.   Collect samples into the Pall eBDS Sample Set as per instructions for use.
21.   Incubate pouches at 35 ± 1°C as indicated in the Pall eBDS Sample Set instructions for
      use.
22.   Measure oxygen concentration in the headspace of the pouch as per Pall eBDS Oxygen
      Analyzer user guide.

Controls: to be run in triplicate
 • Aseptically remove 0.1 mL from tube #6 (containing approximately 1.50 x 103 CFU/mL) and
   spread evenly onto a Trypticase Soy Agar (TSA) with or without blood plate. Incubate at 35
   °C overnight. Conduct colony count on plate to determine starting inoculum. If the McFarland
   standard and subsequent serial dilutions were accurate, there should be approximately 150
   colonies on the TSA plate.
 • After inoculation aseptically remove via syringe and needle 1.0 mL from the inoculated
   platelet product and spread evenly onto a TSA plate. Incubate at 35 °C overnight. Conduct
   colony count on plate to determine bacteria concentration in platelet product. There should be
   approximately 1-15 colonies on the TSA plate.




Last updated 11/22/2006                                                                          16
Vendor/Supplies Information
American Type Culture Collection – Stock Bacteria
PO Box 1549
Manassas, VA 20108
800.638.6597
ATCC.org

Remel – Agar Plates and Broth
12076 Santa Fe Drive
Lenexa, KS 66215
800.255.6730
Remel.org

Dade-Behring – Turbidity Meter
1717 Deerfield Rd.
Deerfield, IL 60015
800.241.0420
Dadebehring.com




Last updated 11/22/2006                             17

				
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