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

e) Most Probable Number (MPN) Analysis. The level of E. coli O157 will be determined by MPN 1 on the day of analysis by evaluating 5 x 650 g, 20 x 325 g (using the reference method test 2 portions) or 5 x 325 g and 5 x 160 g inoculated test samples. Each test portion will be analyzed 3 by the reference method procedure. The number of positives from the 3 test levels was used to 4 calculate the MPN using the LCF MPN calculator available at the AOAC RI website. 5 6 3) 3M™ Molecular Detection Assay 2 - E. coli O157 Method

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a) Applicability

For the detection of E. coli O157 from: raw ground beef (325 g), frozen blueberries (25 g),

sprouts (25 g), fresh baby spinach (200 g).

b) Test Kit Information

a) Kit Name.— 3M™ Molecular Detection Assay 2 – E. coli O157 (including H7)

b) Catalog Number .— MDA2ECO96

c) Test Kit Reagents - 3M MDA2 - E. coli O157 (including H7) (96 tests)

a) Lysis Solution (LS) tubes . – 96 (12 strips of 8 tubes) b) E. coli O157 Reagent tubes . – 96 (12 strips of 8 tubes)

c) Extra caps. – 96 (12 strips of 8 caps) d) Reagent Control (RC). – 16 (2 pouches of 8)

e) Quick Start Guide

d) Additional Supplies and Reagents

a) 3M Molecular Detection System (instrument), power supply, cord, USB cable and software

b) 3M Molecular Detection Speed Loader Tray

c) 3M Molecular Detection Chill Block Tray and Chill block insert d) 3M Molecular Detection Heat Block Insert e) 3M Molecular Detection Cap/Decap Tool [Reagent] f) 3M Molecular Detection Cap/Decap Tool [Lysis]

g) Empty lysis tube rack h) Empty reagent tube rack

AOAC Research Institute Expert Review Panel Use Only

i) 3M™ Enrichment Pouch with 225mL of Buffered Peptone Water (ISO formulation) and filter,

available from 3M #QEBPWI225F

j) Matrix Control, available from 3M #MDMC96NA

e) Additional Media for Confirmation a) Modified Rainbow agar (mRBA)

b) Tryptic soy agar with 5% sheep blood (SBA) c) Dynal antibody coated paramagnetic beads

d) Biochemical confirmation supplies e) Serological confirmation supplies

f)

Apparatus

a) Pipettes . – Capable of 20µL

b) Multi-channel pipette . – Capable of 20µL c) Sterile pipette tips. – Capable of 20µL

DRAFT DOCUMENT

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