Microbiology Methods for ERP Review 3-2020

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GENE ‐ UP ® SLM 2 Collaborative Study

July 2019

Information about this Collaborative Study

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Shipment Schedule 3 4 Test portions will be shipped by overnight express courier to arrive the week before initiation of 5 analysis. If the test portions do not arrive by the normal delivery time (minimum Friday prior to 6 analysis), please contact a co ‐ Study Director. All test portions will be shipped refrigerated. Analysis of 7 test portions must be initiated on the day scheduled by the Study Director.

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Proposed analysis schedule

Food

Date Shipped

Date Tested

Raw Ground Beef (80% lean) 375 g

Thursday TBD

Monday TBD

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Test Portion Receipt 13 14 Note condition of package on the data sheets and notify the Study Director if any packages appear to 15 have been compromised or if the food product does not arrive in the appropriate condition. One 16 temperature probe will be included in the shipment to document the temperature upon receipt. Record 17 temperature on the sample receipt provided in the shipment. All products should be stored at 18 refrigerated temperature (2 ‐ 8 o C) until the day of analysis. 21 22 Collaborators will be provided with enough test portions to test using both the GENE ‐ UP SLM 2 method 23 and the MLG reference method. Collaborators will receive: 36 x 375g test portions (SLM 2) and 36 x 25g 24 test portions (MLG). Please review the reference methods carefully before initiating analysis. 25 26 One additional 60 g control (negative) test portion will be provided to each collaborator for each matrix 27 to determine the total plate count on the day of analysis. Record all results (candidate and reference 28 method; APC) on the data sheet provided (Appendix 8.2). 19 20 Test Portion Analysis 1.2. Aseptically add 1125 mL of pre ‐ warmed BPW to 375 g test portions. Homogenize the sample and incubate at 42 ±1°C for 10 h. After 10 h , a 20 μ L aliquot of each sample is lysed for 5 min at >2000 rpm. Lysed samples are transferred to PCR reaction tubes and assayed on the GENE ‐ UP® 1.3. Regardless of presumptive results, all test portions will be confirmed as if they were positive by the reference method procedure beginning with a transfer to secondary enrichments as outlined in step 2.2. Additionally, an alternative confirmation for the primary enrichment will be conducted by directly streaking the primary enrichment broth to selective agars, CHROMID ® Salmonella and ASAP™, and incubating at 35 ± 1°C for 24 ± 3 h. Final confirmation is performed thermocycler. 1. GENE ‐ UP Salmonella 2 (SLM 2) Method 1.1. Pre ‐ warm bioMérieux buffered peptone water (BPW) to 42 ± 1°C.

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according to step 2.4.

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