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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INTRODUCTION

The goal of this collaborative study is to estimate the following performance parameters of the 3M™ Molecular Detection Assay 2 (MDA2) - E. coli O157 (including H7): probability of detection (POD) and the difference in the POD at 3 contamination levels for both the candidate and reference methods. Test portions from 3 different contamination levels, including negative controls, will be sent to collaborators where they will perform the 3M MDA2 - E. coli O157:H7 method along with the corresponding reference method. Raw fresh

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ground beef will be tested with 325 g test portion sizes.

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Description of the 3M™ MDA2 - E. coli O157 (including H7)

The 3M™ Molecular Detection Assay 2 - E. coli O157 (including H7) is used with the 3M™ Molecular Detection System for the rapid and specific detection of E. coli O157 (including H7) in enriched food, feed and food process environmental samples. The 3M Molecular Detection Assays use loop-mediated isothermal amplification to rapidly amplify nucleic acid sequences with high specificity and sensitivity, combined with bioluminescence to detect the amplification. Presumptive positive results are reported in real-time while negative results are displayed after the assay is completed. Presumptive positive results should be confirmed using your preferred method or as The 3M Molecular Detection Assay 2 - E. coli O157 (including H7) is intended for use in a laboratory environment by professionals trained in laboratory techniques. 3M has not documented the use of this product in industries other than food or beverage. For example, 3M has not documented this product for testing pharmaceutical, cosmetics, clinical or veterinary samples. The 3M Molecular Detection Assay 2 - E. coli O157 (including H7) has not been evaluated with all possible food products, food processes, AOAC Research Institute Expert Review Panel Use Only testing protocols or with all possible strains of bacteria. Summary of Pre-collaborative Study The 3M MDA2 - E. coli O157 method was validated according to AOAC Guidelines (2012) in a pre-collaborative study. The aim of this study was to demonstrate that the 3M MDA2 - E. coli O157 method could detect E. coli O157 in select foods as claimed by the manufacturer. The 3M MDA2 - E. coli O157 was evaluated following the AOAC Official Method requirements: inclusivity/exclusivity and method comparison. The 3M MDA2 - E. coli O157 method was compared to the USDA/FSIS-MLG 5.09 Detection, Isolation and Identification of Escherichia coli O157:H7 from Meat Products and Carcass and Environmental Sponges for raw ground beef (325 g) and to FDA/BAM Chapter 4A Diarrheagenic Escherichia coli for frozen blueberries (25 g), sprouts (25 g) and fresh baby spinach (200 g). Overall the assay was equivalent to the reference methods based specified by local regulations.

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on the Probability of Detection (POD) analysis.

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Study Director for Collaborative Study

DRAFT DOCUMENT

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