#BIOLOGICAL INDICATOR SPORE TESTING PROCEDURE SERIAL#
Make appropriate 10 fold serial dilutions in sterile purified water.Transfer 1 ml aliquots in duplicate to tubes containing 9 ml of sterile purified water.For Bacillus subtilis, heat tube containing the suspension in the water bath at 80oC to 85oC for 10 minutes (heat shock), starting the timing when the temperature reaches 800 C.
For Geobacillus stearothermophillus, heat the tube containing the suspension in the water bath at 95oC to 100oC for 15 minutes (heat shock), starting the timing when the temperature reaches 950 C.Transfer a 10 ml aliquot of the suspension to a sterile, screw-capped test tube.Stir it on vortex mixer for 15 minutes to achieve a homogenous suspension.Open and add the liquid containing BI spores in a sterile 250 ml screw capped bottle containing 100 ml of chilled, sterile purified water.Take three ampoules of the relevant biological indicators.Disperse the strips into component fibers by placing the test specimens in a sterile 250 ml screw capped bottle containing 100 ml of chilled, sterile purified water and mixing.Aseptically remove three specimens of the relevant biological indicator strips from their original individual containers using a sterile forceps.Store as per the manufacturer’s instructions. On receipt of Biological indicators, check for the certificate of analysis, manufacturing date, expiry date, labeled spore count and ‘D’ value.Biological Indicator organisms: Geobacillus stearothermophillus, ATCC 7953 for steam sterilization validation, Bacillus subtilis, ATCC 9372 for dry heat sterilization validation.
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