MPK | STANDING OPERATION PROCEDURE FOR MICROBIOLOGICAL MONITORING OF STERILITY SUIT, SETTLE PLATE METHOD | MPK

 

1.0.      Purpose

To describe the microbiological monitoring (Air microbial count), frequency and procedure of Sterility suit in Microbiology lab through settle plate method, Air sampler and swab method.

STERILITY SUIT

 2.0.      Scope

This procedure is applicable in Microbiology lab of ……… International (Pvt.) Limited.

 

3.0.      Responsibilities

Action	Responsibility
To follow this procedure	Microbiologist
To implement this procedure	Incharge Microbiology Lab

 4.0.     Requirements

·         Tryptic Soy agar

·         Tryptic Soy agar with Lecithin and Polysorbate 80

·         Sterilized Petri plates 90mm

·         70% IPA

·         Sterilized swabs

·         Micro Air Sampler

 

5.0.      Frequency

1.    Settled plate method ------------ Daily

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2.    Through Air sampler ------------  Weekly or as per requirement

3.    Swab method ---------------------- Weekly or as per requirement

6.0.      Procedure

6.1. Settled plate method

6.1.1. Preparation of media plates

 

S. No	Action	Responsibility
6.1.1.1	Aseptically pour the sterilized Tryptic Soy agar to the sterilized Petri plates of 90mm.	Microbiologist
6.1.1.2	Leave the media plates to solidify at room temperature.
6.1.1.3	Incubate these plates at 30 to 35°c for minimum 2 days for pre incubation to eliminate any chance of contamination during preparation.
6.1.1.4	These pre validated plates are used for area monitoring of sterility suit through settled plate method.

    6.1.2. Exposure to sterility suit

S. No	Action	Responsibility
6.1.2.1	Expose media plates in different locations of Sterility suit mentioned in the Annexure-2 for 4 hours.	Microbiologist
6.1.2.2	After exposure, carefully remove the plates and incubate these plates at 30 to 35°c for 48 hrs.
6.1.2.3	After incubation take out the plates and count Microbial colonies. Note down the results in the Annexure-1.
6.1.2.4	If the results are above the acceptable limit then immediately check the HEPA of sterility room and Laminar. Also check cleaning procedures and perform extra cleaning as per SOP.
6.1.3.5	Action limit: As per ISO-14644-2

       6.2. Air sampler

S. No	Action	Responsibility
6.2.1	Prepare the media plates of 90mm and pre validate them as for settled plate method.
6.2.2	Use Air sampler Micro, as per the SOP# S00/QA/M00/P/0201.	Microbiologist
6.2.3	After air sampling, carefully remove the plates from the air sampler and incubate these plates at 30 to 35°c for 48 hrs.
6.2.4	After incubation take out the plates and count Microbial colonies. Note down the results in the Annexure-00.
6.2.5	If the results are above the acceptable limit then immediately check the HEPA of sterility room and Laminar. Also check cleaning procedures and perform extra cleaning as per SOP.
6.2.6	Action limit: As per ISO-14644-2

 

  6.3. Swab method

   6.3.1. Preparation of Swabs with diluents

 

S. No	Action	Responsibility
6.3.1.1	Prepare cotton swabs with clean cotton on the Aluminum sticks.	Microbiologist
6.3.1.2	Prepare 0.1% peptone water and distribute in 2 to 3ml volume in screw capped glass test tubes of size 150x20mm.
6.3.1.3	Immerse the cotton swab in the diluents and cover with the cape. Sterilize these swabs tubes through Autoclave as per SOP # S00/QA/M000/PP/027.

 6.3.2. Sampling from the clean surfaces through sterilize swabs

 

S. No	Action	Responsibility
6.3.2.1	Microbiologist will perform the sampling through sterilized swabs from the tested clean surface in a single move so that it should cover 25x25cm of the area.	Microbiologist
6.3.2.2	Carefully transfers the swabs in the tube containing diluents gently shake it.
6.3.2.3	Mark the tubes with the specified sampled area and carefully take it to the Microbiology lab.

    6.3.3. Microbial testing of sampled swabs through plate count method

 

S. No	Action	Responsibility
6.3.3.1	The number of colony forming units per area will be quantified using plate count method.	Microbiologist
6.3.3.2	Prepare Trypticase media as per SOP # S/Q/M/PP/000.
6.3.3.3	For plate count, aseptically transfer the sampled diluents into the sterilized glass Petri plates under laminar Flow hood and pour cool and melted Trypticase media at 45 to 50°c by gently tilting.
6.3.3.4	Mark these plates as per the sampled area and incubate at 32.5 ± 2.5°c for 48 hrs.
6.3.3.5	After incubation takeout the plates and read them for total Microbial count.
6.3.3.6	Acceptance criteria: As per USP-2009