sop for receipt storage and Determining the population of Biological indicators

1.0 OBJECTIVE
1.1 To describe a procedure for receipt storage and Determining the population of Biological indicators.
2.0 SCOPE
2.1 This procedure applies receipt storage and Determining the population of Biological indicators in Microbiology laboratory
3.0 RESPONSIBILITY
3.1 Officer Microbiologist
3.2 Executive Microbiologist
4.0 ACCOUNTABILITY

4.1 Head –QC
5.0 PROCEDURE

5.1

EZ Test steam self- contained biological indicator
Indicator organism: Bacillus steriothermophillus, ATCC 7953

5.2 Receipt and storage:

5.2.1 On receipt, check for the certificate of analysis, manufacturing date, expiry date and the labeled spore count and entry in register.

5.2.2 Check for the condition of the outer packing. In case of any defect inform the supplier and return it back.

5.2.3 Give the use before date as per the manufacturer’s expiry date.

5.2.4 Store the indicators as per the manufacturer’s instructions.

5.2.5 The ampoule/Strip indicators shall be handled carefully at the time of usage to avoid damage.

5.2.6 The indicators shall be stored and maintained as per the manufacturer’s instructions after receiving until use.

5.3 Identification:

5.3.1 When incubated in Soybean Casein Digest Medium for 17 hrs and under microscopic examination it consists of gram + ve rods with oval endospores. Biochemical tests: Delayed, weak, catalyze positive doesn’t utilize citrate (citrate negative), reduces nitrate, (nitrate reduction positive).

5.4 Population Determination:

5.4.1 Perform the initial count of one strip of B. steriothermophillus spore from the lot/batch
Which are to be used for validation purpose?

5.4.2 Transfer the strip /ampoule aseptically into 100 ml normal saline, keep it for 30 minutes and give heat sock, keep the tube in water bath at 70 0C -80 0C for 15 minutes.
5.4.3 Vortex the tube it to homogenize suspension of the spores.

5.4.4 Depending upon the initial count, dilute further in sterile normal saline.

5.4.5 Take each aliquot in duplicate from different dilutions in such a way that the colony count should not come more than 300 cfu/plate.

5.4.6 Pour the SCDA medium which has been melted & cooled to 45 0C to 50 0Cinto the sterile petriplates, swirl it & let it be solidified.
5.4.7 Incubate the plates in an inverted position at 55 0C – 57 0C for 48 – 78 hrs.

5.4.8 Count the colonies per plate and calculate the average number /strip or ampoule using dilution factor.
5.4.9 Use the strip / ampoule of biological indicator for validation purpose if the ignition count is not more than 150 % of the label claim and not less than 75 % of label claim.
5.4.10 Record the result in format

5.5 Disposal:
5.5.1 Prior to destruction sterilize it by autoclave at 121oC for not less than 30 min.

5.6 Frequency: Whenever a new lot of Biological Indicators are received.

6.0 ABBREVIATIONS
9.1 QCM. : Quality Control Microbiology

9.2 SOP : Standard Operating Procedure
9.3 QA : Quality Assurance
9.4 LAF : Laminar air flow

9.5 CFU : Colony forming unit

sop for calibration and validation of micro autoclave

sop for Sterility failure investigation

cleaning and operation of discard autoclave

sop for operation of fogger machine

sop for Biological assay of lactic acid bacillus

sop for preparation of culture inoculum

STP for sterility testing of sterile gloves