Category: Microbiology SOP

SOP Title: Decontamination of Media, Cultures and Microbial Waste

1. Objective:

To lay down the procedure for Decontamination of Media, Cultures and Microbial Waste.

2. Scope:

This procedure is applicable for Decontamination of Media, Cultures and Microbial Waste in the

Microbiology Section of Quality Control Department.

3. Responsibility:

3.1   Quality Control Department: To prepare and review the SOP. To follow the procedure for Decontamination of Media, Cultures and Microbial Waste as per this SOP.

• Quality Assurance Department: To review and approve the SOP and Annexure.

4. Accountability:

Head Quality Control Department, Head Quality Assurance Department.

5. Procedure:

5.1	Safety Precautions/Instructions:
5.1.1	Ensure that the appropriate protective equipment are used during handling of Autoclave.
5.1. 2	For loading and unloading of autoclave, heat resistant gloves (asbestos free) should be used.
5.1.3	To achieve adequate sterilization, avoid overloading the Autoclave.
5.1.4	Check the water level before each cycle in the Autoclave.
5.1.5	Do not open the Autoclave before the pressure on gauze shows zero.
5.2	Procedure for Decontamination:
5.2.1	Dehydrated media, Broth culture media, Agar media, routine microbiological waste are to be loaded in the Autoclave with self adhesive indicator tape.
5.2.2	Unused dehydrated media quantity to be emptied in the containers and closed securely to prevent any spillage, once the expiry period is over. Before decontamination transfer dehydrated media to Autoclavable container or flask.
5.2.3	All unused broth culture media are collected in Autoclavable container/flask.
5.2.4	Unused Agar media contained in tubes or flasks shall be first heated over boiling water bath to melt the Agar and emptied in a Autoclavable container or flask.
5.2.5	Microbiologically contaminated  culture media e.g. petri dishes, tubes, containers used for  microanalysis for plate count, Raw Materials testing, Finished Product testing and Environmental Monitoring, etc shall be transferred in a autoclavable containers and decontaminated in   autoclave at 121° C and 15 psi for 30 minutes.
5.2.6	These petriplates, tubes, containers marking shall be removed with acetone or methanol or 70% IPA.

 

5.2.7	Transfer dehydrated media, broth and Microbiological waste in a autoclavable containers and decontaminated in   autoclave at 121° C and 15 psi for 30 minutes.
5.2.8	After decontamination pour the contents in Decontaminated Microbiological Waste container and transfer to authorised biomedical waste treatment facility for disposal.
5.2.9	Empty containers, culture tubes, petriplates, flasks are immersed in a container containing 2.5% solution of dettol or savlon for 2 – 3 hours.
5.2.10	Remove and clean the test tubes and flask with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol.
5.2.11	Wash with tap water repeatedly several times and finally rinse with distilled water or purified water and dry at 60°C for 1 -2 hours.
5.3	Assigning Autoclaving Load numbers:
5.3.1	Enter the data in Autoclave Decontamination Record . The Load No. should be given as Follows. eg. DC/001/24. i.e. DC for Decontamination, 001 is first load number for the year and 24 for the Year 2024. This sequence is carried out for one calendar year.
5.4	Cleaning:
	Frequency: Fortnightly
5.4.1	Switch OFF the autoclave.
5.4.2	Disconnect the plug from Mains supply.
5.4.3	Disinfect exterior surface of the Autoclave with 70 % IPA.
5.4.4	Scrub the chamber using 5% v/v Teepol.
5.4.5	Rinse the chamber thoroughly with water to remove all residual particles and detergent.
5.4.6	Rinse finally with purified water.
5.4.7	Wipe the outside of the autoclave with clean cloth.
5.5	Preventive Maintenance:
5.5.1	Perform the preventive maintenance as per Annexure.
5.5.2	If preventive maintenance activity is outsourced, the observations are to be transcribed in the Annexure, and external party certificate to be attached. Frequency of Preventive maintenance is Quarterly.

 

6. Definitions/Abbreviations:

• Definitions :

 

• Abbreviations :

Abbreviation	Expansion
psi	per square inch
%	Percentage
IPA	Iso Propyl Alcohol
e.g.	example
v/v	Volume by volume
ml	milliliter
°C	Degree centigrade
No.	Number

 

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.

 

SOP Title: Procedure for Media Preparation and Sterilisation for various Microbiological Testing

1. Objective:

To lay down a procedure for the Media Preparation and Sterilisation for various Microbiological

Testing.

2. Scope:

The scope of this SOP is applicable for the Preparation of Media, Sterilisation, Dispensing of sterile Culture Media, Storage and Reuse of Sterilized Culture Media, Assigning Batch Number to the Media and the related documentation in Microbiology Section of Quality Control Department.

3. Responsibility:
   • Quality Control Department: To prepare and review the SOP. To follow the procedures laid down for the Procedure for Media Preparation and Sterilisation as per this SOP.
   • Quality Assurance Department: To review and approve the SOP and Annexure.
4. Accountability:

Head Quality Control Department, Head Quality Assurance Department.

 

5. Procedure:

5.1		Procedure for Media  Preparation:
5.1.1		Refer to MSDS of the respective media before handling it for the first time.
5.1.2		Verify that the physical characteristics of the media are as per the Certificate of  Analysis of individual culture media.
5.1.3		Always wear gloves and nose mask while weighing the dehydrated media.
5.1.4		Use a dry spatula for withdrawing media from the container and close the bottle lid tightly after use.
5.1.5		When using a new bottle of dehydrated media enter the date of opening on Media Receipt Label as per SOP ‘Procedure for Receipt and Storage of Dehydrated Culture Media used for Microbiological Analysis’.
5.1.6		Weigh accurately the required quantity of dehydrated culture medium as per Annexure on a butter paper.
5.1.7		Transfer the weighed quantity of media to a suitable flask and reconstitute it with required volume of purified water.
5.1.8		Enter all the details of preparation on “Media Preparation Record” as per Annexure.
5.1.9		Enter the consumption record of Dehydrated Media in usage record as per Annexure.
5.1.10		Update the “Instrument/Equipment Usage Log Book” for balance as per SOP “Calibration of Instruments and Equipments”.
5.1.11		The pH values of the reconstituted media as prepared above should be in compliance with the values indicated on the container label of the dehydrated culture medium or Certificate of analysis.
5.1.12		Check the pH and if necessary, adjust with 0.1 N Hydrochloric acid to decrease the pH value or 0.1 M Sodium Hydroxide solution to increase pH.
5.1.13		Record the pH of reconstituted media in Annexure and update the “Instrument/Equipment Usage Log Book” for pH as per SOP “Calibration of Instruments and Equipments”.
5.1.14		Agitate briskly for a few minutes to dissolve or suspend the adherent powder to form a uniform slurry or solution. Avoid spillage.
5.1.15		Place the flask on boiling water bath and occasionally shake up to complete dissolution of media or a suspension.
5.1.16		Transfer the media in flasks or tubes as per the requirement with appropriate plugging either with non absorbent cotton or other suitable material such as plastic caps.
5.1.17		Broth media should be dispensed before sterilisation directly in the test tubes to be used for analysis.
5.2		Procedure for Sterilisation of Media:
5.2.1		Check the water level of autoclave. Ensure that the electric supply is off during media loading and unloading.
5.2.2		For operation procedure of autoclave, refer SOP ‘Procedure for Operation and Cleaning of Autoclave’.
5.2.3		Place the sterilisation indicator strip with media container and affix it as a reference after sterilisation on Annexure, ‘Autoclave Sterilization Record’ of SOP, ‘Procedure for Operation and Cleaning of Autoclave’.
5.2.4		Transfer the flasks/tubes containing reconstituted media to the autoclave with proper plugging. Avoid overloading.
5.2.5		Tightly close the lid of autoclave and sterilise the media at 121°C +2°C at 15 psi for 15 minutes or as specified in Annexure.
5.2.6		Record the autoclave cycle number on Annexure as well as on Annexure of ‘Autoclave Sterilization Record’ of SOP, ‘Procedure for Operation and Cleaning of Autoclave’ as a reference after sterilisation.
5.2.7		After sterilisation unload the media using the heat resistant gloves (asbestos free) to prevent burns.
5.2.8		Transfer the reconstituted media to the inoculation room through the pass box for dispensing and solidification.
5.2.9		Check the pH of sterile media by taking a small quantity of media in a test tube or in suitable glassware under aseptic conditions and make the entry in Annexure.
5.2.10	Record the pH of reconstituted media in Annexure and update the “Instrument/Equipment Usage Log Book” for pH as per SOP “Calibration of Instruments and Equipments”.
5.3	Procedure for Dispensing of Sterile Culture Media:
5.3.1	Molten agar media should be poured around 15-20 ml in sterile Petri plates and 5-10 ml if preparing the slants under the Laminar Air Flow, when cooled to about 45 °C.
5.3.2	Allow the media to solidify at room temperature.
5.3.3	Record the balance quantity of prepared media along with its validity period in Annexure.
5.3.4	Label the flasks and test tubes of 100 ml volume or above as per Annexure ‘Reconstituted Media Label’.
5.3.5	In case of petri plates and test tubes of 30 ml volume, mark the Batch No. and the Date of preparation manually with a marker pen.
5.4	Procedure for Storage and Reuse of Sterilized Culture Media:
5.4.1	1.   The petri plates/slants shall be stored in the refrigerator or at room temperature.
5.4.2	2.   Prepared media shall be used for a maximum period of 15 days from the date of preparation or as per the SOP ‘Procedure to Determine Shelf Life of Reconstituted Media.’
5.4.3	3.   Remove the stored agar media from refrigerator before use and allow it to stand to attain the room temperature.
5.4.4	4.   Re-melt the solid agar media on the day of use in the original container on the water bath at 100° C.
5.4.5	5.   The media usage record shall be entered in Annexure.
5.4.5	6.   Stored culture media showing any change in appearance or any signs of visible microbial contamination or cracks or dehydration shall be discarded as per SOP ‘Procedure for destruction of media, cultures and microbial waste’.
5.4.6	7.   All used glassware’s shall then be cleaned as per SOP ‘Procedure for Cleaning of Used Glassware in Microbiology Section’.
5.5	Assigning Batch Number to the Media
5.5.1	Allocate each reconstituted media with an individual batch number.
5.5.2	Batch numbering shall be done as Media code/Serial number/Year; wherein
5.5.3	Media code specifies about the unique code defined for each dehydrated media on its receipt.  Refer Annexure, In-house Code of Dehydrated Media, Bases and Reagents of SOP, ‘Procedure for Receipt and Storage of Dehydrated Culture Media used for Microbiological Analysis’
5.5.4	Serial number is the batch number allotted to each reconstituted media.
5.5.5	Year specifies the last two digits of calendar year.
5.5.6	For example; M1/001/24 stands for the batch number of reconstituted media of MacConkey agar, where; M1 : Media code of MacConkey Agar. 001: First batch of reconstituted MacConkey Agar media. 24  : Last two digits of calendar year.
5.5.7	Record the batch number of reconstituted media in Annexure of Media Preparation and Usage Record of Agar/Broth Medium.
5.5.8	Unused solid media to be discarded by melting in water bath after its expiry.
5.5.9	Decontaminate all the used Media by referring procedure given in SOP, Decontamination of culture media and microbial waste.

 

6. Definitions / Abbreviations:

• Definitions :

 

• Abbreviations :

Abbreviation	Expansion
°C	Degree Centigrade
0.1 N	0.1 Normal
0.1 M	0.1 Molar
ml	millilitre
psi	Per square inch
+	Plus
MSDS	Material Safety Data Sheet

  

*Note – Ready to use SOP available in “DOWNLOAD” Section.

 

 

 

 

SOP Title: Cleaning of Used Glassware in Microbiology Section

1. Objective:

To lay down the written procedure for Cleaning of Used Glassware in the Microbiology Section of

Quality Control Department.

2. Scope:

This procedure is applicable for Cleaning of Glassware and other utensils used for microbiological

analysis in the Microbiology Section of Quality Control Department.

3. Responsibility:

• Quality Control Department: To prepare and review the SOP. To follow the procedures laid down for Cleaning of Used Glassware in the Microbiology Section in Quality Control Department as per this SOP.
• Quality Assurance Department: To review and approve the SOP and Annexures.

4. Accountability:

Head Quality Control Department, Head Quality Assurance Department.

5. Procedure:

5.1	Precautions:
5.1.1	All contaminated glassware’s containing culture media and other biohazards must be sterilised in the autoclave at specified conditions, before cleaning to kill all the hazardous microorganisms.
5.1.2	Always wear personal protective equipments such as safety goggles, gloves, nose masks during washing of glassware’s.
5.1.3	Do not use brushes that are so worn that the spine hits the glass as a result serious scratches may result and scratched glass is more prone to break during analysis.
5.1.4	Do not drop glassware’s from height or bang to the sink or tap.
5.1.5	Visually check glassware’s for any cracks before washing. Broken glassware’s may cause injury.
5.1.6	Do not shake glassware’s too much vigorously as it may get cracks by pressure.
5.1.7	Do not collect or touch broken glass pieces with bare hands. It can cause injury.
5.1.8	Ensure that the soap has been thoroughly removed from the glassware because soap when comes in contact with the acid can form a film of grease.
5.1.9	Do not mix the glassware’s from the microbiology section with the chemical section since it may cause cross contamination and impact the media component as well as the growth of microorganisms.
5.1.10	Protect cleaned glassware’s from dust by placing them in a dust free cabinets.

 

5.2	General Procedure:
5.2.1	Laboratory technician should wear safety goggles, gloves, masks before starting the washing of glassware’s.
5.2.2	Microbiologist must decontaminate the media and other hazardous microbial waste prior to cleaning, as per the SOP, Procedure for destruction of cultures and microbial waste (used culture media).
5.2.3	Drain off any non hazardous contents present in the flasks, beakers, test tubes etc. in the sink and the hazardous contents that have been decontaminated in the autoclave, in the waste containers labelled as ‘Decontaminated Microbiological Waste’.
5.2.4	Rinse the glassware’s with tap water before transferring them into soap solution.
5.2.5	It is the Microbiologist’s responsibility to drain off hazardous contents from the flask, petriplates or any other glassware before giving it for washing.
5.2.6	Immerse the glassware’s in tub containing 2.5% solution of dettol or savlon for 3-4 hours.
5.2.7	Remove the markings of pen marker on glassware’s with the methanol.
5.2.8	Remove the silicon grease on glassware’s with Acetone or use fuming sulphuric acid (Oleum) for about 30 minutes and then rinse with purified water.
5.2.9	Scrub inner and outer portions of glassware’s with scrubber or nylon brushes soaked in 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran.
5.2.10	If thorough cleaning is not possible immediately, soak the glass wares in 1% v/v Teepol solution or any other soap solution till adhesive material is removed or dissolved.
5.2.11	During washing all parts of glassware’s should be thoroughly scrubbed with nylon brush.
5.2.12	Rinse the glassware’s with running tap water; allow the water to run into and over the glassware’s for short time until no foam of soap is produced on shaking.
5.2.13	All the glassware’s should be finally rinsed with purified water twice and kept in tilted position to drain off water completely.
5.2.14	Dry the glass wares in the glassware drying oven at 60°C ±5°C.
5.2.15	New glassware’s should be soaked in 1% v/v solution of hydrochloric acid or nitric acid for about 5-6 hours before washing as new glassware’s are slightly alkaline in reaction.
5.2.16	If glassware becomes unduly cloudy  or dirty or contains coagulated organic matter then glass wares should be given chromic acid treatment as follow:-
5.2.17	Chromic acid solution: Dissolving 84g of chromium trioxide in 700 ml water and add slowly, with stirring 400 ml of sulphuric acid. (While preparing solution use personal protective equipments such as goggles, gloves, a mask as chromic acid is classified as a carcinogen).
5.2.18	Pour the few ml of chromic acid solution on dirty glassware’s and leave them for minimum 15 minutes. Add small amount of potable water with shaking.
5.2.19	Rinse several times with water or allow to keep in water for about one hour as the chromic acid is highly acidic in nature and finally rinse with purified water twice.
5.2.20	Dry the glass wares in the glassware drying oven at 60°C ±5°C.
5.2.21	Use the washed glassware within one week of the washing. After one week re-wash the same glassware’s before use even if they are not used.
5.2.22	Store the washed and dried glassware in a designated and clean cupboard with proper labelling.
5.3	Cleaning of Test tubes, Durham’s tubes and Centrifuge tubes:
5.3.1	Empty the contents of the test tubes in a container labelled as ‘Decontaminated Microbiological Waste’.
5.3.2	Durham’s tube can be retrieved while transferring the contents of test tube in the container labelled as ‘Decontaminated Microbiological Waste’.
5.3.3	Wash the tubes under the tap and immerse the tubes in tub labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3-4 hours.
5.3.4	Remove and clean the tubes with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran.
5.3.5	Wash with tap water until all traces of soap solution are removed.
5.3.6	Finally rinse with distilled water or purified water and keep in a tilted position to drain off water completely.
5.3.7	Dry the tubes in drying oven at 60°C ± 5°C to remove the moisture.
5.3.8	Sterilise the tubes before use, as per the SOP, Procedure for operation of sterilisation autoclave.
5.3.9	Test tubes for culture maintenance should be cleaned and stored separately.
5.4	Cleaning of Petri plates:
5.4.1	Remove the melted media subsequent to sterilisation from the petriplates and empty in a container labelled as ‘Decontaminated Microbiological Waste’ and wash under the tap water.
5.4.2	Immerse the petri plates in tub labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3-4 hours.
5.4.3	Remove and clean the petri plates with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. Use a scrubber for cleaning.
5.4.4	Wash with tap water until all the traces of soap solution are removed.
5.4.5	Finally rinse with distilled water or purified water and keep in an inverted position to drain off water completely.
5.4.6	Dry the petri plates in drying oven at 60°C ± 5°C to remove the moisture.
5.4.7	Sterilise the petri plates before use, as per the SOP, Procedure for operation of sterilisation autoclave.
5.5	Cleaning of Pipette:
5.5.1	Immerse the pipette in a container labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3 – 4 hours.
5.5.2	Ensure that the level of disinfectant in the container is high enough to immerse the maximum portion of the pipettes.
5.5.3	Do not drop them into the container. This may break the tips and render the pipettes useless for accurate measurements.
5.5.4	Blocked Pipette should be immersed in a cylinder filled with dichromate cleaning solution and left for 24 hours.
5.5.5	Remove and clean the pipettes with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran.
5.5.6	Wash with tap water until all the traces of soap solution are removed.
5.5.7	Finally rinse with distilled water or purified water.
5.5.8	Dry the pipettes in drying oven at 60°C ± 5°C to remove the moisture.
5.5.9	Sterilise the pipettes before use, as per the SOP, Procedure for operation of sterilisation autoclave.
5.6	Cleaning of Flask for media preparation:
5.6.1	Empty the contents of the Flasks in a container labelled as ‘Decontaminated Microbiological Waste’.
5.6.2	Wash under the tap and immerse the flasks in tub labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3-4 hours.
5.6.3	Remove and clean the flasks with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran.
5.6.4	Wash with tap water until all the traces of soap solution are removed.
5.6.5	Finally rinse with distilled water or purified water and keep in a tilted position to drain off water completely.
5.6.6	Dry the flasks in drying oven at 60°C ± 5°C to remove the moisture.
5.6.7	Sterilise the pipettes before use, as per the SOP, Procedure for operation of sterilisation autoclave.
5.7	Cleaning of Dropper pipette, tips for micropipette:
5.7.1	Wash under the tap and immerse the dropper pipettes and tips in a container labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3-4 hours.
5.7.2	Remove and clean the dropper pipettes and tips with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran.
5.7.3	Wash with tap water to remove all traces of soap solution.
5.7.4	Finally rinse with purified water.
5.7.5	Dry the dropper pipettes and tips in drying oven at 60°C ± 5°C to remove the moisture.
5.7.6	Tips for micropipette can be used for analysis, after sterilisation as per the SOP, Procedure for operation of sterilisation autoclave.

 

5.8	Cleaning of Microscopes Slides and Cover slips:
	1.   New Slides:
5.8.1	Place slides overnight in a beaker of 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran.
5.8.2	Rinse each slide under tap water until all traces of soap solution are removed.
5.8.3	Wipe the slides, one at a time with a soft cloth. Place on blotting paper or filter paper that doesn’t shed any fibres and allow to dry.
5.8.4	Pack the slides in stacks of 10 or 20 groups in paper.
	2.   Dirty slides:
5.8.5	Oil is removed by rubbing with blotting paper. One or two drops of alcohol can be used to remove residual oil.
5.8.6	Cover slips are removed using a needle or forceps.
5.8.7	Now soak in beaker of 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran.
5.8.8	If slide have been used for infected specimen, they should be placed in disinfectant solution of 2.5% solution of dettol or savlon for 3-4 hours.
	3.   Cover slips:
5.8.9	Immerse the cover slips in a beaker of 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran.
5.8.10	Soak for 2-3 hours, shaking gently from time to time.
5.8.11	Rinse the beaker with cover slips under tap water.
5.8.12	Finally rinse with distilled water or purified water.
5.8.13	Drain the cover slips out on a gauze pad.
5.8.14	Dry in hot air oven at 60°C till drying and keep in a clean petri dish.

 

6. Definitions/Abbreviations:

• Definitions :
• Hazardous waste: A hazardous waste is a waste that poses substantial or potential threats to health or the environment.
• Non-hazardous waste: Non-hazardous waste means the waste that is not harmful to health and the environment.
• Carcinogens: A substance or agent that can cause cells to become cancerous by altering their genetic structure so that they multiply continuously and become malignant.
• Sterilization: Sterilizationis a term referring to a process that eliminates or kills all forms of life, including transmissible agents (such as fungi, bacteria, viruses, spore forms, etc.)

 

• Abbreviations :

Abbreviation	Expansion
SOP	Standard Operating Procedure
QC	Quality Control
QA	Quality Assurance
%	Percent
v/v	Volume by volume
°C	Degree centigrade
g	grams
ml	Milliliter

 

 *Note – Ready to use SOP available in “DOWNLOAD” Section.

 

SOP Title: Entry Exit Procedure in Microbiology Section

1. Objective:

To lay down the written procedure for Entry Exit in Microbiology Section of Quality Control

Department.

2. Scope:

This procedure is applicable for Entry Exit of working personals and visitors in the Microbiology

Section of Quality Control Department.

3. Responsibility:

• Quality Control Department: To prepare and review the SOP. To follow the procedures laid down for Entry Exit Procedure in Microbiology Section of Quality Control Department as per this SOP.
  • Quality Assurance Department: To review and approve the SOP and Annexures.
• Accountability:

Head Quality Control Department, Head Quality Assurance Department.

5. Procedure:

5.1	Precautions:
5.1.1	The Good Laboratory Practices must be read prior to entering the microbiology section. By being knowledgeable about the procedures, analyst will assure their own safety.
5.1.2	Person must sanitize their hands before entering the microbiology section and after working with potentially hazardous materials, before leaving the laboratory.
5.1.3	Never wear open toed shoes because of the constant danger of cuts and infections from broken glass found on the lab floors and the possibility of chemical or microbial culture spills.
5.1.4	Decontaminate work surfaces after completion of work and after any spill or splash of potentially infectious material with appropriate disinfectant.
5.1.5	Do not take microbial cultures out of microbiological area since it may cause fatal infections if not handled properly.
5.1.6	Ensure that single door is opened at a time. Do not open the next one until the previous one has been closed, to prevent the cross contamination.
5.2	Entry Procedure for the Microbiology Section:
5.2.1	Entry should be restricted in Microbiology Laboratory for other than concern personnel.
5.2.2	Switch ‘ON’ the department lights.
5.2.3	Check the differential pressure on magnehelic gauge is minimum 1mm before entering in Microbiology Laboratory.
5.2.4	Before starting the work, ensure that the cleaning activities are completed in the presence of the house keeping personnel/head.
5.2.5	A person entering in the microbiology section of the Quality Control Department should enter in first cubical by pushing the door and remove the company footwear and disinfect hands with 70% IPA and should enter in second cubical by pushing the door and wear the sterilised over gown available in the box provided at second cubicle along with the sterile hand gloves, mask and headgear. Ensure that minimum part of the body gets exposed within the microbiology section. One over gown can be used for the day by the same person.
5.2.6	Enter the third cubical. Disinfect the hands with disinfectant available in third cubical before entering the inoculation room. Push the door with the reverse side of the palm to prevent contamination.
5.2.7	Verify the HVAC working condition by monitoring the temperature and Humidity of the section. If not, intimate the engineering department.
5.2.8	Incubation room entry is provided through first cubical of change room. Push the door of incubation room and enter inside.
5.2.9	There is no need of over gowning while entering in the media preparation room and incubation room.
5.2.10	Protective nose mask, safety goggle, hand gloves and heat resistant gloves are required for autoclave handling and weighing of dehydrated media.
5.3	Exit Procedure for the Microbiology Section:
5.3.1	Disinfect the hands before leaving the inoculation room with 70 % IPA available in inoculation room, enter in third cubicle and disinfect hands with disinfectant available in the third cubicle, then enter into the second cubicle, remove the over gown and other accessories and keep it in the rack provided. Used hand gloves should be discarded in Dustbin after every use. Then enter First cubicle, disinfect hands with 70 % IPA and then exit.
5.3.2	After the Analysis is completed for the day, remove the used over gown and keep it in a container labelled as “Linen for washing” available in first cubicle to send it for washing at the end of shift.
5.3.3	At the end of the analysis ensure that all the instruments/equipments used are switched ‘OFF’, except the ones which are to be kept ‘ON’ for 24 hours such as incubators. Also clean the work benches before leaving the department.
5.3.4	Used glassware’s shall be disinfected and cleaned as per SOP ‘Procedure for Cleaning of Used Glassware in Microbiology Section’. Broken glassware to be discarded into the box marked “Broken Glassware” and not into waste bins.
5.3.5	Ensure the valve of gas cylinder used in inoculation room and the regulator valve of gas cylinder in microbiological section are closed, as mentioned in SOP ‘Procedure for operation of laminar flow work table’.
5.3.6	Verify that all the chemicals, media, reagents, liquid solvents are properly closed and kept in their respective places after use and the water supply taps are also closed.
5.3.7	During exit all doors should be opened and closed one by one, same as that of entry procedure.
5.3.8	Ensure that lights are switched ‘OFF’ before leaving the section.
5.3.9	All visitors should be accompanied by authorized company employee in Microbiology Section of Quality Control Department.
5.3.10	All visitors should follow the same entry and exit procedure as mentioned above.

 

6. Definitions/Abbreviations:

• Definitions :

 

• Abbreviations :

Abbreviation	Expansion
SOP	Standard Operating Procedure
QC	Quality Control
QA	Quality Assurance
HVAC	Heating, Ventilating and Air Conditioning
LAF	Laminar Air Flow
LPG	Liquid Petroleum Gas

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.