Standard Procedure For AUTOCLAVE QUALITY CONTROL by uif20236

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									                         Saint Louis University – Biological Safety Committee
                                        Standard Procedure
                                                  For
                               AUTOCLAVE QUALITY CONTROL
                                     (adapted December 1997; updated May 13, 2005)

{Investigators submitting applications to the Biological Safety Committee must commit to following this protocol or
                                          submit an equivalent procedure}

PRINCIPLE
  The ability of the autoclave to sterilize is checked with a live-spore preparation (ATTEST). If
  adequate sterilization is achieved, then the spores fail to germinate.

SPECIMEN
  Not applicable.

REAGENTS/EQUIPMENT
  1. ATTEST vials containing spores of the thermophilic organism Geobacillus
     stearothermophilus are used (3M Corporation).
  2. Autoclave.
  3. 56°C water bath (i.e., ATTEST incubator or equivalent)

STORAGE
  1. Store ATTEST biological indicators under normal room conditions: 59 - 68°F (15 - 30°C,
     35-60% relative humidity.
  2. Do not store indicators near sterilants or other chemicals.
  3. ATTEST indicators have a two-year shelf life from the date of manufacture. The
     manufacturing date is printed on the ATTEST biological indicator label, box and shipping
     carton.

DISPOSAL
  Dispose of used ATTEST biological indicators in accordance with the healthcare facility’s
  policy. Any positive indicators may be autoclaved at 250°F (121°C) for at least 15 minutes or
  at 270°F (132°C) for ten minutes in a gravity steam sterilizer, or for four minutes in a pre-
  vacuum or pulsing steam sterilizer.

PROCEDURE
  1. The minimum frequency of using test vials is weekly.
  2. The vial should be placed in the center of the autoclave in the middle of other material
     being autoclaved.
  3. Run autoclave.
     CAUTION: After sterilization, the contents of ATTEST biological indicators are hot
                    under pressure. Always allow to cool before handling. Failure to cool
                    for at least ten minutes may cause the glass ampule to burst which may
                    result in injury from flying debris.
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  4. After the vial has cooled, crush the glass ampule by applying pressure with a pencil or
     other hard object. Wear gloves when doing this. As a control, crush the glass ampule of an
     unautoclaved vial.
  5. Incubate an unautoclaved vial and an autoclaved vial in a 56°C water bath for two days.
     The vials must be incubated within two hours after the glass ampule is crushed.
  6. Examine the vials at 24 and 48 hours for a color change. Appearance of a yellow color
     indicates bacterial growth. Growth should occur in the unautoclaved vial while no growth
     should be obtained in the vial that was autoclaved.
  7. Record results on a worksheet, such as that included as an attachment to this procedure,
     that you will post near the autoclave.
  8. Record the temperature of the ATTEST incubator when vials are placed in the ATTEST
     incubator and at 24 and 48 hours when examining the vials
  9. If the autoclaved vial is culture positive, perform the following:
     a. Repeat the test with new vials. Do not use autoclave until QC testing indicates no
          growth of spore solution.
     b. If the autoclaved vial is again positive, check temperature and pressure gauge.
     c. If no malfunction is discernible, call the biomedical department.
     d. Do not use the autoclave until repair is completed and a repeat sterility test results in no
          growth of the spore solution.

LIMITATIONS OF PROCEDURE
  1. Culture media must still be tested for sterility since media do not require the length of
     exposure that is required for sterilization of spore solutions.

  2. Maintaining the temperature of the water bath at 56°C (133°F) is critical to the
     accuracy of the procedure.
  3. Failure to maintain a proper water level in the ATTEST incubator can result in a
     temperature in excess of 56°C (133°F).

REFERENCES

  Rutala, W.A. 1995. In: Murray, et al. Manual of Clinical Microbiology, 6th Edition. ASM,
  Washington, D.C., p. 240




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     BIOLOGICAL MONITORING FOR STEAM STERILIZATION USING ATTEST


Autoclave Identification: ___________________________ Location: _____________________

Culture   Date of Run   Time     Length of             Test Vial       Control Vial
 No.                           Autoclave Cycle      24          48     24       48
                                                   Hours      Hours   Hours    Hours
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