AOAC RI ERP Final Action Recommendations eBook

Collaborative Study Approved Protocol Expert Review Panel Use Only

For the analysis of orange juice, collaborators will receive 72 test portions, 36 test portions to be analyzed by the BAX® System Real-Time PCR Assay for Salmonella and 36 test portions to be analyzed by the FDA/BAM reference method. Each set of 36 test portions will contain: 12 un-inoculated controls, 12 low level (target 0.2 – 2.0 cfu/test portion), and 12 high level (target 2 – 5 cfu/test portion). For the analysis of hot dogs, collaborators will receive 36 test portions. The test portions will be analyzed using the BAX® System Real-Time PCR Assay for Salmonella and the USDA/FSIS MLG reference method. See flow diagrams 8.4 and 8.5. Test portions of each matrix will be inoculated with a different strain of Salmonella spp . , randomized and blind-coded for analysis. See Table 1 below for summary of the evaluation.

Table 1. Overview of Study Design

Test Week (Tentative Date)

# Test Portions (per method)

Target Salmonella Levels 0 cfu/test portion 0.2-2 cfu/test portion 2-5 cfu/test portion 0.2-2 cfu/test portion 2-5 cfu/test portion 0 cfu/test portion

Total Test Portions/Week

Reference Method

Matrix

12

24

Orange Juice 25g –BAX Real- Time 25g – BAM Hot Dogs 325g –BAX Real-Time and MLG

Week 1 (January 14, 2013)

FDA/BAM Chapter 5

12

24

12

24

12

12

Week 2 (January 22, 2013)

USDA/FSIS MLG Ch. 4.05

12

12

12

12

• Each laboratory will prepare and analyze all test portions. All test portions will be randomized and blind-coded with a 3 digit reference number.

• Collaborators will also receive one portion of each food matrix labeled “Temperature Control”. Temperatures will be taken and recorded on the data sheet provided upon receipt of samples. One additional 60 g APC 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 (Friday) by the pour plate method. Please note that the Temperature Control and the APC Control are not test samples and will not be artificially contaminated.

• One matrix will be tested each week of the study. See below for shipping schedule.

• Q Laboratories will prepare the test portions and ship them refrigerated via Federal Express Overnight Priority to each collaborator. The first set of sample test portions,

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