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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d) Serological Pipette Bulbs (Automatic Pipette) – For sampling and delivering of 1 mL - 10 mL

e) Serological pipettes . – Aerosol resistant Stomacher® . – Seward or equivalent g) Filter Stomacher® bags . – Seward or equivalent f)

h) Sterile collection sponge and swab . – Environmental surface sampling i) Thermometer . – Calibrated range to include 100 ± 1°C range j) Incubators . – Capable of maintaining 35±2°C, 42±1°C and 41.5±1°C k) Dry Bath Incubator . – Capable of maintaining a temperature of 100 ± 1 ºC

l) Dry double block heater unit. – Capable of maintaining 100 ± 1°C; or a water bath capable of

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maintaining 100 ± 1°C

m) Refrigerator . – capable of maintaining 2-8°C, for storing the 3M MDA2 n) Computer . – compatible with the 3M Molecular Detection System (instrument)

g) Safety Precautions

The 3M™ Molecular Detection Instrument is intended for use with samples that have undergone heat treatment during the assay lysis step, which is designed to destroy organisms present in the sample. Samples that have not been properly heat treated during the assay lysis step may be considered a potential biohazard and should NOT be inserted into the 3M Molecular

Detection Instrument.

The user should read, understand and follow all safety information in the instructions for the 3M Molecular Detection System and the 3M MDA2 – E. coli O157 (including H7). Retain the safety

instructions for future reference.

h) Sample Preparation

a) Foods - 3M recommends the use of 3M BPW ISO for the enrichment of food samples. It is the user’s responsibility to validate alternate sampling protocols or dilution ratios to ensure

this test method meets the user’s criteria.

(1) Pre-warm BPW ISO enrichment medium to 41.5 ± 1°C.

(2) Aseptically combine the enrichment medium and sample according to information below. For all meat and highly particulate samples, the use of filter bags is AOAC Research Institute Expert Review Panel Use Only recommended. (a) Ground beef – 325g + 975 mL BPW ISO (pre-warmed to 41.5 ± 1°C). Homogenize the sample for 2 ± 0.2 min. Incubate at 41.5 ±1°C for 10-18 hr. (b) Sprouts – 25g + 225 mL 3M BPW ISO (pre-warmed to 41.5 ± 1°C ). Homogenize the sample for 2 ± 0.2 min. Incubate at 41.5 ±1°C for 18-24 hr. (c) Frozen whole blueberries – 25g + 225 mL 3M BPW ISO (pre-warmed to 41.5 ± 1°C ). Gently agitate the sample for 2 ± 0.2 minutes (DO NOT STOMACH). Incubate at 41.5 (d) Spinach – Add 200g + 450 mL BPW ISO (pre-warmed to 41.5 ± 1°C ) in a sterile re- sealable plastic bag and gently agitate by hand. Incubate at 41.5 ±1°C for 18-24 hr. (e) Run the 3M Molecular Detection Instrument following the 3M Molecular Detection ±1°C for 18-24 hr.

System User Manual – Reserving enriched test portion for confirmation.

i)

Post-enrichment

DRAFT DOCUMENT

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