Procedure for autoclave validation

•     OBJECTIVE

•          To lay down a procedure for autoclave validation.

• SCOPE
  • This SOP is applicable for autoclave validation in microbiology laboratory .
• RESPONSIBILITY
  • Officer / Executive-QC – responsible for Implementation of SOP.
• ACCOUNTABILITY
  • Manager – Quality Control
• PROCEDURE
  • Method-A:

•          Clean and load the autoclave for sterilization with accessories and glassware’s.
•          Prepare the required number of (10ml) tubes of Soya bean casein digest medium.
•         Take require nos. of strips of spores of Geobacillus stearothermophilus ATCC 7953.
•          Put in petriplates and    label it, arrange the petiplates at different location no.
•          Keep one strip unexposed for positive control.
•         Run the cycle at 15 lbs. pressure and 121^o C temperature for 15 min.
•         After completion of cycle remove all tubes and Petri dishes.
•         Mark location no.,date on all the SCDM  tube .
•          Transfer strips aseptically to respective labelled tubes.
•          Transfer one unexposed strip to one SCDM tube and label it as positive control.
•          Keep one SCDM tube uninoculated and label it as negative control.
•         Incubate all the tubes at 55^o C ±2^o C for 7days
•          At the end of 7 days all the the inoculated SCDM tubes should show no growth.
•          Negative control tube should show no growth.
•         Positive control tube should show growth with in 72 hrs.
•        The above observations will demonstrate satisfactory validation of the autoclave.
•        Record the observation in Format-I.

•   Method-B

•        Validation is done by the bioindicator system consists of ampoule containing spores of    Geobacillus stearothermophillus ATCC 7953.
•       These ampoules are stored under refrigerator when not in use. On the day of validation
•        Take three ampoules and allow them to come at room temperature.
•        Load the autoclave and place the ampoule in different locations in autoclave within glass beakers to avoid accidental breakage of ampoule during sterilization.
•        Run the sterilization cycle as per procedure. (validated sterilization condition 121°C for 15 min  at 15 PSI.)
•       Unload the chamber and remove the ampoule, mark them and incubate at 55°C ±2°C for     48 hour.
•       The third non sterile unexposed ampoule is incubated along with the sterilized ampoule as a Positive control.
•        At the end of 48 hour days.
•        Positive control – Original purple colour gradually changes to deep yellow with turbidity.
•        Exposed ampoule – The Original purple colour remains as it is at the end of incubation period.
•       The above observations will demonstrate satisfactory validation of the autoclave.
•       Record the result in Format-I

•    ABBREVIATION

•   SOP      –  Standard operating procedure
• IPA       –  Iso Propyl Alcohol
•   SCDM     – Soybean Casein Digest Medium

•     FORMAT

•    DISTRIBUTION LIST

                                                      Format -I

• Equipment Detail:

•  Detail of Test:

•  Details of Validation:

• Details of Observation:

•  Acceptance Criteria:

There should not be any growth in form of turbidity in any of the tubes except Positive control.

•  Corrective action if observation does not meets the requirements:

•  Conclusion:

•  Approval:

OBSERVATION SHEET

•                                  Format -II

       Log Book of Sterilization(Autoclave) Load Number  

Instrument I.D.: ______________________