INSTRUCTIONS FOR HANDLING AND ANALYSIS

A. Receipt, Handling & Storage

Precaution: It is expected that participating laboratories ensure PT items are handled by trained and authorized personnel who possess the necessary competence and awareness regarding the specific handling procedures, safety precautions related to the test parameters, and the hazards associated with the chemicals involved. Should any uncertainty arise during the handling of PT items, please contact the designated personnel at +91-9167702413 or via email at [email protected] for clarification.

Please note below points prior to initiate the analysis:

• Upon receipt of the PT item/sample, confirm if the sample is intact & immediately inform the PTP if needed.
• Record the date of receiving and store properly till the analysis is performed.
• Initiate the analysis within 48 hours of receiving the sample.
• The test materials should be treated in the same way as routine samples and should therefore be tested using your routine laboratory equipment, reagents, and methods.
• The entire content of the vials & container must be utilized for analysis.
• Read all the instructions carefully before analysis.

Note: Participants are expected to use the specified method(s) mentioned below (Reporting of Results) or any other validated methods based on these specified methods.

B. Instructions for Analysis: Specific

For: 2025MHT01-1

1. Uncap the sample vials 2025MHT01-1 aseptically and rehydrate using 4ml sterile saline peptone water (0.1% peptone with 0.85% NaCl).
2. Recap securely & allow 5 minutes at room temperature for rehydration.
3. Aseptically transfer 1 g of the PT item matrix from container 2025MHT01-1 to the sterile enrichment broth for P aeruginosa. Homogenize this.
4. Transfer 2 ml of rehydrate content from vial 2025MHT01-1 into the homogenized broth prepared in step 3, ensuring thorough homogenization of the sample.
5. proceed with your routine analysis for P.aeruginosa.
6. Aseptically transfer 1 g of the PT item matrix from container 2025MHT01-1 to the sterile enrichment broth for S.aureus. Homogenize this.
7. Transfer remaining 2 ml of the rehydrated vial content from 2025MHT01-1 to the sterile dilluent prepared in step 6, ensuring thorough homogenization.
8. proceed with your routine analysis for S.aureus.

For: 2025MHT01-2

1. Uncap the sample vials 2025MHT01-2 aseptically and rehydrate using 5ml sterile saline peptone water (0.1% peptone with 0.85% NaCl).
2. Recap securely & allow 5 minutes at room temperature for rehydration.
3. Aseptically transfer the 1 g of the sample from 2025MHT01-1 to the sterile enrichment broth for Salmonella and Homogenize this.
4. Transfer 2ml of rehydrate vial content into the diluent prepared in step 3, ensuring thorough homogenization of the sample.
5. Proceed with your routine analysis for Salmonella.
6. Aseptically transfer 1 g of the PT item matrix from container 2025MHT01-1 to the sterile dilluent for E.coli. Homogenize this.
7. Transfer remaining 2 ml of the rehydrated vial content from 2025MHT01-1 to the sterile dilluent prepared in step 6, ensuring thorough homogenization.
8. proceed with your routine analysis for E.coli.

For: 2025MHT01-3

1. Uncap the sample vials 2025MHT01-3 aseptically and rehydrate using 5ml sterile saline peptone water (0.1% peptone with 0.85% NaCl).
2. Recap securely & allow 5 minutes at room temperature for rehydration.
3. Aseptically transfer the 10 g of the PT item matrix from 2025MHT01-1 to the sterile bag/bottle/flask and add 85 ml dilluent. Homogenize this.
4. Empty the rehydrate vial content into the diluent prepared in step 3, ensuring thorough homogenization of the sample.
5. Perform serial dillution for 10-1 to 10-5 and proceed with your routin analysis for Total Plate Count.

For: 2025MHT01-4

1. Uncap the sample vials 2025MHT01-4 aseptically and rehydrate using 5ml sterile saline peptone water (0.1% peptone with 0.85% NaCl).
2. Recap securely & allow 5 minutes at room temperature for rehydration.
3. Aseptically transfer the 10 g of the PT item matrix from 2025MHT01-1 to the sterile bag/bottle/flask and add 85 ml dilluent. Homogenize this.
4. Empty the rehydrate vial content into the diluent prepared in step 3, ensuring thorough homogenization of the sample.
5. Perform serial dillution for 10-1 to 10-5 and proceed with your routin analysis for Yeast and Mould.