Working Guideline in Microbiology Laboratory & its SOP

Working Guideline in Microbiology Laboratory; Purpose

To confirm that microbiological best laboratory practice and special precautions are maintained in Microbiology laboratory in order to prevent laboratory & personal contamination.

Working Guideline in Microbiology Laboratory; Scope

This SOP applies for maintaining microbiological best laboratory practices and special precautions in Microbiology Laboratory at XX Pharmaceuticals Limited.

Definitions/ Abbreviation

None

Responsibilities:

The roles and responsibility is as follows

 Executive/ Senior Executive, Microbiology

Maintain microbiological best laboratory practices & special precautions.

Manager, Microbiology

Ensure microbiological best laboratory practices, special precautions and application of sound technical information.

Head of Quality Assurance

Take initiative to Approval of SOP

Procedure:

Media Preparation and Quality Control

Working Guideline in Microbiology Laboratory; Media Preparation

• Choose correct media or components in making media based on the use of accepted sources or references for formula.
• Check Certificate of Analysis describing expiry date, storage conditions etc.
• Read instructions on the label carefully before media selection and preparation.
• Follow if any special instruction such as heating, additives & pH adjustment etc.
• Use always purified water for media preparation.
• Record accurate weight of media & make up volume with purified water.
• Dissolve media thoroughly into water prior to dispensing & sterilization.
• Avoid overheating or less heating media.
• Sterilize the media as per label of the container.

Media Storage

• Label media properly with batch or lot numbers, preparation & expiry dates.
• Store media according to manufacturer’s instructions.
• Store prepared media under validated conditions.
• Do not store agar at or below 0⁰ Because freezing could damage the gel structure.
• Protect stored media from exposure to light & excessive temperature.
• Before prolonged storage, place into a sealed package or container to retard moisture loss.
• Re-melt agar media in a hot water bath or by using free flowing steam only once.

Quality Control Testing of Media:

• Check pH & growth promotion to confirm media efficacy.
• Perform limited growth promotion test for each lot, if media is sterilized using a validated method.
• Do not use media if any parameters do not comply.
• Pre-incubate & 100% inspection prior to use the media.
• Maintain double-wrapped condition for those media used in critical environmental monitoring.

Maintenance of Microbiological Cultures:

• Handle with care microbiological cultures due to their pathogenicity and toxicity.
• Use as standard culture from international recognized organization such as American Type Culture Collection (ATCC), National Collection of Type Culture (NCTC) or any other manufacturer.
• Select Standard culture as freeze dried lyophilized condition in vial or ampoules form or ready to use condition with Certificate of Analysis.
• Ensure identification of culture prior to its use.
• Use only permitted method for culture maintenance.
• Maintain storage and sub-culture as per standard operating procedure for standard culture maintenance.
• Be care to prevent extreme sub-culturing working control cultures that increase the risk of contamination.
• Preserve all typical culture at 2 to 8⁰ 
• Inoculate frozen stock monthly or weekly.
• Discard any unused portion to minimize risk of loss of viability & contamination of the stock.

Maintenance of Laboratory Equipment

• Perform standard validation practices such as Installation Qualification, Operational Qualification and Performance Qualification for equipment.
• Calibrate equipment periodically.
• Check performance of all equipment’s on a routine basis.
• Maintain always approved protocol for IQ, OQ & PQ of equipment’s.
• Clean & disinfect all equipment’s as per standard operating procedure for that equipment.

Laboratory Operation

• Isolate items between sterility testing room & microbial limit test and bio-assay room.
• Do not interchange item between those area without sterilization.
• Sanitize carefully both hands with appropriate sanitizer.
• Before entrance into the test room, wear garments, mask and gloves as appropriate.
• Always use sterile garments, gloves and mask into sterility testing area.
• Aseptically handle the items into microbial limit test room.
• Disinfect all items with 70% IPA or any others suitable sanitizer, before transfer the items into testing room. Open growth plate only into bio-assay room or microbial testing room.
• Aseptically collect the sample.
• Identify contaminants at least genus level.
• Use specific sample area to preserve all samples.
• Isolate all contaminated samples to reduce the false-positive results.
• Follow SOP for disposal of used media to discard all contaminants.
• Clean & disinfect all area according to SOP of cleaning and sanitizing Quality Assurance Department.
• Lessen the movement into microbial testing room and bio-assay room.
• After completion of sterility, exit from sterility testing room.
• Wear another set of sterile dress, gloves, mask If require to re-enter into sterility test room.
• Wash both hands with liquid soap or any skin cleanser solution, after completion of test and before exit from Laboratory, Finally disinfect both hands.

Documentation

• Prepare documents in approved format.
• Preserve documents into specific file up to defined period.
• Review & update all standard operating procedure, specification & analytical Method within defined period.
• Verify the data & calculation.

Maintenance of Laboratory Records

• After completion of each work, prepare record in approved format
• Follow approved SOP to reflect how the test is actually performed.
• Keep calibration record after Perform calibration of any equipment’s
• Preserve all records after approval.
• Do not discard any approved record without permission and proper justification.
• Forward test result of raw materials, in-process & finished products to the respected department.
• Achieve and protect all laboratory record against catastrophic loss.

Spillage Management

Spills:

• Report immediately to the reporting authority after spillages of culture.
• Do not touch any spilled cultures & surrounding debris( e. g. glass , cotton or wool plugs) with exposed hands.
• Wear disposable gloves & disinfect area by covering spill with several layers of paper towel/ cloth soaked in a suitable disinfectant.
• Leave for it for 15-30 minutes.
• Using paper towels, sweep spill debris into dustpan.
• Transfer all disposable materials to a suitable container e.g. an roasting bag/autoclave for autoclaving and disposal.
• Decontaminate dustpan either by autoclaving or by soaking (at least 24 hours) in hypochlorite solution.

Broken glass

• Sweep carefully into a suitable container.
• Dispose in a puncture proof container.

Splashes on clothing and skin

• Soak in disinfectant contaminated cloth.
• Treat to splash on skin as soon as possible.
• Wash thoroughly with soap & finally with hot water.
• Disinfect the skin if necessary.