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The document outlines the protocol for performing growth promotion tests on Trypcase Soy Agar plates to certify new lots of the media. It describes testing two plates of the new lot against two plates from a previously certified lot and calculating the percentage recovery of bacteria/yeast colonies. It also details receiving, storing, sampling, inoculating and incubating the plates, observing and documenting results to determine if the lot passes certification.
The document outlines the protocol for performing growth promotion tests on Trypcase Soy Agar plates to certify new lots of the media. It describes testing two plates of the new lot against two plates from a previously certified lot and calculating the percentage recovery of bacteria/yeast colonies. It also details receiving, storing, sampling, inoculating and incubating the plates, observing and documenting results to determine if the lot passes certification.
The document outlines the protocol for performing growth promotion tests on Trypcase Soy Agar plates to certify new lots of the media. It describes testing two plates of the new lot against two plates from a previously certified lot and calculating the percentage recovery of bacteria/yeast colonies. It also details receiving, storing, sampling, inoculating and incubating the plates, observing and documenting results to determine if the lot passes certification.
Document No. Effective date Version No. Page No. Page 1 of 8 Prepared by Name:
Sign & Date Approved by: Name: Sign & Date
I. Objective: Growth promotion test is performed on microbiological culture media to assess the ability of the media to sustain growth. This protocol outlines the procedure for Growth promotion testing at SPT. The micro-organisms used are recommended by the European Pharmacopoeia & USP.
II. Scope: The scope of this protocol is applicable to the Growth Promotion test, certification, usage and reconciliation of Trypcase Soy Agar (TSA) plate received at MICRO lab. III. Procedure: Once a new lot of media from an approved vendor is received, it is necessary to certify the lot. The lot may be certified by performing growth promotion tests. Documentation of growth promotion tests will be done in this document including corresponding certificate of analysis and usage and reconciliation. Each lot will have separate protocol / report. The report will be completed and archived on exhaustion of specific lot of the media. Growth promotion test is performed by comparing one plate of new lot of media to two plates of previously certified lot. A certified lot is a lot of agar of known fertility. In-case of initial lot, this shall not be applicable. The colony forming unit (CFU) count in the new lot of media is then compared to the mean count of the certified plates and the percentage recovery is calculated. A. Determination of microorganisms that are needed to test the medium: Medium Organism Incubation temperature Incubation time Trypcase soy Agar (Tryptone Soya Agar) Staphylococcus aureus ATCC 6538 - represent as gram positive Escherichia coli ATCC 8739 - represent as gram negative Candida Albicans ATCC 10231 - represent as yeast 30 o C to 35 o C
30 o C to 35 o C
30 o C to 35 o C 3 days
Protocol for Growth Promotion Test Trypcase Soy Agar Plates (Tryptone Soya Agar) Document No. Effective date Version No. Page No. Page 2 of 8
IV. Receipt, verification and storage of TSA Plates: Name of the product (as mentioned on Certificate, PO and product label)
Supplier Catalogue No. Batch No.
Mfg. Date
Exp. Date Quantity received Purchase Order reference number & date Supplier Storage condition CoA availability Pack integrity check Satisfactory / Not satisfactory
Details of damage / remarks (if any):
Received and verified by: Name:
Signature and date:
Final remarks of consignment Accepted / Rejected On satisfactory verification, apply quarantine label on the pack and store at respective storage conditions / location.
V. Sampling Instruction: Two plates to be tested for growth promotion tests. Two plate to be used a negative control.
Protocol for Growth Promotion Test Trypcase Soy Agar Plates (Tryptone Soya Agar) Document No. Effective date Version No. Page No. Page 3 of 8 Prepared by Name:
Description Escherichia coli ATCC 8739 Lot No.: Candida Albicans ATCC 10231 Staphylococcus aureus ATCC 6538 Date & time of preparation Performed by (Sign & date) Date & time of preparation Performed by (Sign & date) Date & time of preparation Performed by (Sign & date) Remove the hydrating fluid vial and vial of lyophilized strain preparation from refrigerated storage. Allow unopened vial of lyophilized pellets to equilibrate to room temperature
While pellets are equilibrating, pre-warm the hydrating fluid to 35 o C (at least 30 minutes)
With a sterile forceps, transfer two (2) pellets into the 2 ml vial of hydrating
Protocol for Growth Promotion Test Trypcase Soy Agar Plates (Tryptone Soya Agar) Document No. Effective date Version No. Page No. Page 4 of 8
Description Escherichia coli ATCC 8739 Lot No.: Candida Albicans ATCC 10231 Staphylococcus aureus ATCC 6538 Date & time of preparation Performed by (Sign & date) Date & time of preparation Performed by (Sign & date) Date & time of preparation Performed by (Sign & date) fluid. (do not remove the desiccant from vial). Two pellets must be used to obtain the challenge concentration of <100 CFU per 0.1 ml. Immediately stopper and recap the pellet vial and return the remaining lyophilized material to refrigerated storage of 2 o C to 8 o C Immediately re-cap the vial with the hydrated material and place into a 34 o C to 38 o C incubator for thirty (30) minutes to assure complete hydration
Immediately following incubation vortex the hydrated material to achieve a homogeneous suspension
VI. Testing Procedure:
Protocol for Growth Promotion Test Trypcase Soy Agar Plates (Tryptone Soya Agar) Document No. Effective date Version No. Page No. Page 5 of 8
Description Specification / procedure Observation Done by (Sign & date)
Date of testing (start date): Date completed (after observation of results):
Label plates with the organism name, duplicate number, date of inoculation and media type To comply
Using sterile pipette, pipette-out 0.1 ml of each microorganism solution to the media plates To comply Volume transferred:
Lot No. of sterile Pipette:
Volume transferred:
Lot No. of sterile Pipette:
Volume transferred:
Lot No. of sterile Pipette:
Using sterile spreader spread the solution evenly across the surface of the agar Allow plates to dry before inverting and incubation. To comply Lot no. of sterile spreader:
Lot no. of sterile spreader:
Lot no. of sterile spreader:
Two agar plate (of new lot) incubated along with the test plates without any inoculation. This will act as negative control
To comply Record the time and date of incubation period in the Growth Promotion Test Report of individual medium.
To comply Incubator No.:
Date Incubated:
Time Incubated:
Incubator No.:
Date Incubated:
Time Incubated:
Incubator No.:
Date Incubated:
Time Incubated:
After incubation, remove all plates from the incubator, record the time and date that the To comply Date removed:
Date removed:
Date removed:
Protocol for Growth Promotion Test Trypcase Soy Agar Plates (Tryptone Soya Agar) Document No. Effective date Version No. Page No. Page 6 of 8
Description Specification / procedure Observation Done by (Sign & date)
Inspect each plate for growth, counting all CFU present on each plate, The percentage recovery is calculated using the formula given below:
For New Lot
Average Test count (new lot) (CFU) % recovery = ----------------------------------------- X 100 Average count of approved plates
The average count must be between 10 to 100 CFU.
% recovery must not differ by a factor greater than 2 (i.e., 50% to 200%)
No growth must be observed in negative control New lot: Plate 1:
Plate 2
Average:
Previously approved lot: Plate 1:
Plate 2:
Average:
% recovery:
Negative control observation:
New lot: Plate 1:
Plate 2
Average:
Previously approved lot: Plate 1:
Plate 2:
Average:
% recovery:
Negative control observation:
New lot: Plate 1:
Plate 2
Average:
Previously approved lot: Plate 1:
Plate 2:
Average:
% recovery:
Negative control observation:
Protocol for Growth Promotion Test Trypcase Soy Agar Plates (Tryptone Soya Agar) Document No. Effective date Version No. Page No. Page 7 of 8 Prepared by Name:
Sign & Date Approved by: Name: Sign & Date
VII. Certificate of Analysis Name of the Media Delivery Note No. Batch No. / Lot No. Quantity received Mfg Date Quantity approved Expiry date Quantity rejected Date of receipt Date of Certification Organism Specification / results Remarks (Pass / Fail) Average test count % recovery Escherichia coli ATCC 8739
Candida Albicans ATCC 10231
Staphylococcus aureus ATCC 6538
Review remarks of supplier CoA To confirm the purity and authenticity of supplied culture
Acceptance Criteria: Average test count: Not More Than 100 CFU; test plate count must not differ from control plate count by factor greater than 2 Remarks: The received consignment Complies / Does not complies to the specifications. The material is approved / rejected for further intended use Prepared by (Signature and date) Approved by (Signature and date)
Protocol for Growth Promotion Test Trypcase Soy Agar Plates (Tryptone Soya Agar) Document No. Effective date Version No. Page No. Page 8 of 8
VIII. Usage log: Stock control and reconciliation (use additional sheets where needed) Quantity Released Product Name / Item B. No. Test Name Date used Quantity used