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.