(Certified Strength 10 6)ĥ.3 Perform the initial count of one strip of B.steriothermophilus spores from the lot/batch which are to be used for validation purpose.ĥ.4.1 Transfer the strip aseptically into 100 ml sterile Normal Saline, keep it for 1/2 hr and give heat shock, keep the tube in water bath at 70☌-80☌ for 15 minutes.ĥ.4.2 Shake the tube by palming or vortex it to homogenise suspension of the sporesĥ.4.3 Depending upon the initial count, dilute further in sterile Normal Salineĥ.4.4 Take each aliquot in duplicate from different dilutions in such a way that the colony count should not come more than 300 cfu/plate.ĥ.4.5 Pour the SCD agar medium which has been melted & cooled to 45☌ to 50☌ into the allocated plates, swirl it & let it be solidified.ĥ.4.6 Incubate the plates in an inverted position at 55 ± 2☌ for 48-72 hours.ĥ.4.7 Count the colonies per plate and calculate the average number/strip using dilution factor.ĥ.4.8 Use the strip for validation purpose if the initial count is not more than 150 % of the label claim and not less than 75% of the label claim.ĥ.5.1 Load the autoclave chamber with accessories and glassware.ĥ.5.2 Take requires nos. This procedure is applicable for Autoclave installed in microbiology area of quality control laboratory.ģ.1 Doing: Tech.Assistant (Microbiologist)/Executiveĥ.2 Organism: Bacillus steriothermophilus spores. ![]() To validate autoclave with a biological indicator for sterilization efficiency.
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