AOAC RI ERP Final Action Recommendations eBook

Collaborative Study Approved Protocol Expert Review Panel Use Only

Additional Supplies and Reagents required but not provided 18. Pipettes – For transferring reagents; two adjustable mechanical pipettes covering 20-200 µ L and 5-50 µ L; one repeating pipette; and one multi-channel pipette covering 8 channels and 5-50 µ L. Pipettes should be calibrated to deliver required volumes within 10%. 19. Pipette tips with barriers – 0.5-250 µ L, 0.5-100 µ L extended barrier; 5 mL repeater pipette tips. 20. PCR tube holders – For transferring a rack of tubes from the cooling block to the cycler/detector. 21. Incubators – For maintaining enrichment media at 35 ± 1 ° C and 39-42 ° C. 22. Cluster tubes and racks – For lysis. 23. Capping/de-capping tools – For removing and sealing cluster tube caps and PCR tube caps without jarring the contents. 24. Thermal block(s) with inserts – For maintaining lysis tubes at 37 ± 2°C, 95 ± 3°C, and 4 ± 3°C. 25. Water bath - Capable of maintaining 42 ± 1°C for secondary selective enrichments. 26. Refrigerator – capable of maintaining 2-8°C, for storing the BAX® System Real-Time PCR Assay for Salmonella test kits 27. Stomacher® – Seward model 400 or equivalent. 28. Stomacher® type bags or equivalent. 29. Blender – High speed (16,000 – 18,000 rpm) with sterile receptacle.

* Note: Please ensure that ample incubator space is available to accommodate the larger test portions. We estimate that each lab will need approximately 54 cubic feet (3 large incubators or an incubator room) to facilitate the testing of the 325 gram test portions. Overfilling the incubator with too many test samples may compromise the integrity of the results.

Test Portion Analysis

Collaborators will be provided with enough test material to test each matrix using the BAX ® System Real- Time PCR Assay for Salmonella. For orange juice, they will receive two sets of test portions: 36 x 25g (BAX ® System) and 36 x 25g (FDA/BAM). For hot dogs, collaborators will receive one set of test portions: 36 x 325g (BAX ® System and USDA/FSIS MLG). One temperature control sample will also be included in the shipment to document the temperature of the sample upon receipt. One additional 60 g control (negative) test portion will be provided to each collaborator for each matrix to determine the total plate count on the day of sample receipt. Record all results on the data sheets provided. See Appendix 10.3 for data sheets. Please review the reference method carefully before initiating analysis.

Reference Pre-enrichment

BAX ® System Enrichment

Food Type

Reference

BPW (Paired Enrichment)

BPW (Paired Enrichment)

MLG 1

Hot Dogs

31