AOAC-RI ERP Book Micro Jan 19 2017

OMAMAN-35 B: Collaborative Study Protocol ERP USE ONLY January 2017

3M MDA2 E. coli O157:H7 Collaborative Study

May 2016

OMA-2016-May-XXXX

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Shipment Schedule 2 3 Test portions will be shipped by overnight express courier to arrive the day before initiation of analysis. 4 If the test portions do not arrive by the normal delivery time one day prior to analysis, please contact a 5 co-Study Director. All test portions will be shipped refrigerated. Analysis of test portions must be 6 initiated on the day scheduled by the Study Director. Test kits will be sent directly from 3M.

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Proposed analysis schedule

Food

Date Shipped

Date Tested

Raw ground beef 325 g

Thursday

Monday

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Test Portion Receipt 12 13 Note condition of package on the data sheets and notify the Study Director if any packages appear to 14 have been compromised or if the food product does not arrive in the appropriate condition. All products 15 should arrive cold to the touch and should be stored at 4  C (refrigerated) until the day of analysis. 18 19 Collaborators will be provided with enough test portion to test using the 3M MDA2 - E. coli O157 20 (including H7) method and the reference method. For raw ground beef testing, collaborators will 21 receive: 36 x 325g (3M) and 36 x 325g (MLG). See Appendix 8.5 for detailed instructions on analysis. 22 Please review the reference methods carefully before initiating analysis. 23 24 One additional 60 g control (negative) test portion will be provided to each collaborator for each matrix 25 to determine the total plate count on the day of analysis. One temperature control sample will also be 26 included in the shipment to document the temperature of the sample upon receipt. Record all results on 27 the data sheet provided. 28 29 1. 3M Molecular Detection Assay 2 - E. coli O157 (including H7) Method 30 AOAC Research Institute Expert Review Panel Use Only 16 17 Test Portion Analysis 1.1. Pre-warm 3M buffered peptone water BPW ISO enrichment medium to the incubation temperature: 41.5 ±1°C. For meat and highly particulate samples, filter bags are recommended. 1.2. Aseptically add 325 g test portion to 975 mL BPW ISO. Homogenize the sample for 2 ± 0.2 min. Incubate at 41.5 ±1°C. After 10 hr , pull a sample from ground beef enrichment to run the 3M Molecular Detection Instrument following the 3M Molecular Detection System User Manual –

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Reserving enriched test portion for confirmation.

1.3. Confirmation of alternative method - All samples, regardless of presumptive result, must be

confirmed starting with step 2.3 below. 38 2. USDA/FSIS Reference Method, MLG online Chapter 5.09 – E. coli O157:H7 39

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2.1. Add 325g into 975 mL mTSB. Stomach each test portion for approximately two minutes.

Incubate at 42  1  C for 15-24 h.

2.2. Perform the screening test for E. coli O157:H7/non-motile following the manufacturer

instructions for the screening method listed in USDA FSIS MLG 5A.

DRAFT DOCUMENT

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