Growth Promotion Test In Microbiology Laboratory
1.0 OBJECTIVE
1.1 To lay down the procedure for Growth Promotion Test to check the fertility of the procured culture media.
2.0 SCOPE
2.1 This procedure applicable for Growth Promotion Test In Microbiology Laboratory
3.0 RESPONSIBILITY
3.1 Microbiologist QC
4.0 ACCOUNTABILITY
4.1 Q. C. Manager
5.0 PROCEDURE
5.1 PRECAUTIONS
5.1.1 The receipt of microbiological media, its entry should be done in media stock records with details such as Batch Number,
Quantity received, Expiry date, Date of opening, Date of issue, Bottle No. issued, Balance quantity, Opened and checked by.
5.1.2 The microbiological media is stored in an appropriate place of storage as per instruction given on container.
5.1.3 Each new lot of microbiological media shall be qualified for Growth Promotion Test.
5.2 MATERIAL REQUIRED
5.2.1 Culture medium, Normal saline (0.9%), Petri plates, Inoculating loop, Sterile L- spreader, Media Test Tube, Micropipette (100-1000µl), Micro tips (100-1000µl),& Microbial culture.
5.3 EQUIPMENT REQUIRED:
5.3.1 HLAF, Autoclave, BOD Incubators (32.5˚C ± 2.5˚C & 22.5˚C ± 2.5˚C, 43˚C ± 1˚C).
5.4 Methodology
5.5 Preparation of culture suspension
5.5.1 Take a freshly prepared culture which should not be more than 48 hrs. old.
5.5.2 Harvest the complete microorganism growth in normal saline tube.
5.5.3 Prepared the culture suspension as per SOP
5.6 Growth Promotion Test for Agar Media
5.6.1 Prepare the required agar media as per SOP for Media preparation.
5.6.2 Spread on the media plates by sterile spreader not less than 0.1 ml of culture suspension containing not more
than 100 cfu for each organism with the help of Micropipette.
5.6.3 Use at least two Petri plates for each medium and each of the dilution.
5.6.4 Incubate all the plates at temperature and time as per required by the organism.
(Refer incubation table ).
5.6.5 Also put a blank media plate as negative control and incubate at appropriate temperature and
time.
5.6.6 Calculate the mean count and number of cfu observe the plates and note down the results in
record.
5.7 Growth Promotion Test for Media broth
5.7.1 Prepare the media as per SOP for Media preparation .
5.7.2 Add 1.0 ml of culture suspension containing not more than 100 cfu per ml in media test tube.
with the help of 1000 µl Micropipette.
5.7.3 Label a blank media tube as negative control.
5.7.4 Incubate the tubes as per time and temperature specified for the organism. The incubation
time for negative control does not less than 5 days. (Refer incubation table).
5.7.5 Observe the tubes for turbidity and note down the results in the record.
5.8 Inhibitory Properties of Liquid and Solid media
5.8.1 Prepare the media as per SOP for Media preparation .
5.8.2 Inoculate the appropriate medium with not less than 100 cfu of the appropriate microorganism
5.8.3 Incubate at the specified temperature for the time specified in the test. (Refer incubation table ).
5.8.4 There should not be any growth of the microorganism on this medium.
5.9 Indicative properties
5.9.1 Prepare the required agar media as per SOP for Media preparation .
5.9.2 Spread on the media plates by sterile spreader not less than 1.0 ml of culture suspension containing not
more than 100 fu for each organism with the help of Micropipette.
5.9.3 Incubate all the plates at temperature and time as per required by the organism.
(Refer Incubation table).
5.9.4 Also put a blank media plate as negative control and incubate at appropriate temperature and time.
5.9.5 Compare the appearance of Colony morphology and indication reaction, obtained with previously
tested and approved batch/Lot of medium.
5.10 Observation – The test media is considered satisfactory, if definite microbial growth
i.e. colonies or turbidity appears in all the containers with in the period specified.
5.11 The negative control does not show any presence of microbial growth.
Note:- % Recovery of the test media should not differ by a factor greater than 2 from the standard inoculums used.
| Name of media
to be used |
Microorganism to be used For testing | Microorganism used For Inhibitory /Indicative | Incubation
Temp |
Incubation
Period |
|
Fluid Thioglycollate medium |
Ps. aeruginosa | – | 32.5 + 2.5 | 48 to 72hrs |
| S. aureus | – | 32.5 + 2.5 | 48 to 72hrs | |
| Cl.Sporogenes | – | 32.5 + 2.5 | 48 to 72hrs | |
| Environment isolates | – | 32.5 + 2.5 | 48 to 72hrs | |
| Soyabean Casein Digest medium
(For Sterility) |
Bacillus subtilis | – | 22.5 + 2.5 | 72to120hrs |
| C .albicans | – | 22.5 + 2.5 | 72to120hrs | |
| A. brasiliensis | – | 22.5 + 2.5 | 72to120hrs | |
| Soyabean Casein Digest medium
(For Water Testing) |
Ps. aeruginosa | – | 32.5 + 2.5 | 48 to 72hrs |
| S. aureus | – | 32.5 + 2.5 | 48 to 72hrs | |
| E coli | – | 32.5 + 2.5 | 48 to 72hrs | |
| Cl.Sporogenes | – | 32.5 + 2.5 | 48 to 72hrs | |
| Environment isolates | – | 32.5 + 2.5 | 48 to 72hrs | |
| C. albicans | – | 22.5 + 2.5 | 72to120hrs | |
| A.brasiliensis | – | 22.5 + 2.5 | 72to120hrs | |
| Sabouraud Dextrose Agar | C. albicans | – | 22.5 + 2.5 | 72to120hrs |
| A.brasiliensis | – | 22.5 + 2.5 | 72to120hrs | |
| Cetrimide agar base | Ps. aeruginosa | E.coli | 32.5 + 2.5 | 48 to 72hrs |
| R.V. S. Enrichment Broth | Salmonella abony | S. aureus | 32.5 + 2.5 | 48 to 72hrs |
| Xylose Lysine Deoxycholate agar | Salmonella abony | E.coli | 32.5 + 2.5 | 48 to 72hrs |
| MacConkey agar | E. coli | 32.5 + 2.5 | 48 to 72hrs | |
| MacConkey broth | E. coli | S. aureus | 32.5 + 2.5 | 48 to 72hrs |
| Mannitol Salt agar | S. aureus | E. coli | 32.5 + 2.5 | 48 to 72hrs |
| Enterobacteria enrichment broth mossel | E. coli | S. aureus | 32.5 + 2.5 | 48 to 72hrs |
| P. aeruginosa | S. aureus | 32.5 + 2.5 | 48 to 72hrs | |
| Violet Red Bile Glucose Agar | E. coli | P.aeruginosa | 32.5 + 2.5 | 48 to 72hrs |
|
New B.No. Received of SCDA & SCDM |
P. aeruginosa | – | 32.5 + 2.5 | 48 to 72hrs |
| S. aureus | – | 32.5 + 2.5 | 48 to 72hrs | |
| E. coli | – | 32.5 + 2.5 | 48 to 72hrs | |
| S. abony | – | 32.5 + 2.5 | 48 to 72hrs | |
| Bacillus subtilis | – | 32.5 + 2.5 | 48to72 hrs | |
| Cl.Sporogenes | – | 32.5 + 2.5 | 48 to 72hrs | |
| Environment isolates | – | 32.5 + 2.5 | 48 to 72hrs | |
| C. albicans | – | 22.5 + 2.5 | 72to120hrs | |
| A.brasiliensis | – | 22.5 + 2.5 | 72to120hrs | |
|
R2A Agar |
Ps. aeruginosa | – | 32.5 + 2.5 | 48 to 72hrs |
| S. aureus | – | 32.5 + 2.5 | 48 to 72hrs | |
| E coli | – | 32.5 + 2.5 | 48 to 72hrs | |
| S. abony | – | 32.5 + 2.5 | 48 to 72hrs | |
| B.subtilis | – | 32.5 + 2.5 | 48 to 72hrs |
5.12 FREQUENCY:
Growth promoting every autoclave lot media. Inhibitory and indicative properties of media whenever new media lot received
and end the container. In case of Soyabean Casein Digest Agar perform growth performance test at receiving time.
6.0 ABBREVIATIONS
| Sr. No. | Abbreviation used | Full form of abbreviation used |
| 1.0 | SOP | Standard Operating Procedure |
| 2.0 | IPA | Iso Propyl Alcohol |
| 3.0 | QA | Quality assurance |
| 4.0 | SCM | Soyabean Casein Digest Medium |
| 5.0 | IP | Indian Pharmacopeia |
| 6.0 | HLAF | Horizontal Laminar Air Flow |
| 7.0 | 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
sop for Operation and calibration of active air sampler
sop for transfer of material for testing and sampling in sterile area
entry & exit procedure in microbiology laboratory
Growth Promotion Test In Microbiology Laboratory
Operation of B.O.D in Microbiology Laboratory
Operation of Horizontal Laminar Air Flow in the microbiology laboratory
Operation and cleaning of Pass Box.
Operation and cleaning of air sampler
Cleaning and Sterilization of Glassware
Analysis of water for microbial load in microbiology lab
Operation and temperature monitoring of Refrigerator
Fumigation of Microbiology Laboratory.
Entry & Exit procedure In Sterility Area
SOP for Microbial analysis of Raw Material Finished Products
SOP for Operation & Calibration of pH meter in Micro Department
SOP for Operation & Calibration of pH meter in Micro Department
SOP Operation and calibration of Hot Air Oven In Microbiology
SOP for operation cleaning & calibration of Digital colony counter
SOP for Operation And Cleaning of Microscope
sop for Media Preparation and Consumption
sop for Receipt Storage and Usage of Culture Media
sop for Cleaning Sanitization And Disinfection In Microbiology
sop for Environmental monitoring of all the Classified area
sop for Handling and Sub culturing of Microbial cultures
sop for Media Growth Promotion Test and various Microbiological test
sop for BOD incubator operation and cleaning
sop sampling of water for microbiological analysis
sop for Disinfectant Efficacy Test
sop for for cleaning and operation of vortex mixture
sop for Temperature & Relative Humidity Monitoring
sop for Operation and Calibration of Heating Block
sop for Sterility Testing of Microbiology
sop for Disposal of Culture Media
sop for Drain point of Microbiology
sop for entry & exit procedure In Microbial limit test and Biosafety
sop for Gram Staining of Bacteria in Microbiology Laboratory
sop for Monitoring of Compressed Air/gases for microbiological analysis
sop for BET (Bacterial Endotoxin) test in Microbiology
sop for receipt storage and Determining the population of Biological indicators
sop for qualification of analyst microbiologist
sop for Bioburden test of Packing materials in Microbiology Laboratory
sop for microbiological assay of erythromycin antibiotic
sop for liquid particle counter
sop for operation and calibration of digital zone reader
sop for monitoring of ultraviolet efficiency LAF and pass box
microbiological assay of cyanocobalamin or vitamin B12
gowning procedure for microbiological testing area
swab testing of various surfaces for bioburden determination
sop for endotoxin challenge test
Hold time study protocol for sterilized media
sop for personnel Qualification protocol for aseptic area
sop for sampling and testing of drain water
Sop for Operation of Airborne Particle Counter
sop for Validation protocol of steam sterilizer autoclave
sop for pathogen detection from drain point
Sop for Analysis of Raw water Purified water water for injection and pure steam water
sop for preservatives efficacy test
sop for collection and preservation of in house isolated microorganisms
sop for Operation Calibration and Maintenance of Micropipette
sop for microbiological testing of water
sop for depyrogenation of apparatus
sop for fertility test growth promotion test of media
sop for Operation and cleaning of moist heat sterilizer
sop for monitoring by active air sampler
sop for swab sampling and testing for clean rooms in production area
sop for monitoring in microbiology laboratory
sop for Fumigation of aseptic area and microbiology lab
sop for monitoring of personnel in aseptic area
sop for maintenance of cultures
sop for Operation and cleaning of laminar bench
sop for monitoring of pure steam
sop for entry and exit procedure to m.l.t and b.e.t room
sop for storage of and use of media
sop for disposal of microbiological media and cleaning of microbiological glassware