AOAC-RI ERP Micro June 2016

OMAMAN-29/OMAMAN-30 Combined Collaborative Study Protocol OMA ERP - June 2016 ERP Use Only

3M MDA 2 Listeria species and L. monocytogenes Collaborative Study OMA-2016-MONTH-XXX

5.3.9 Dey-Engley (D/E) Neutralizing Broth 5.3.10 Demi-Fraser Broth

Additional media 5.4.1 Fraser broth 5.4.2 Oxford Agar (OX)

5.4

5.4.3 Modified oxford agar (MOX) 5.4.4 Horse blood overlay agar (HL) 5.4.5 Sheep blood agar (SBA) Apparatus 5.5.1 Pipettes . – capable of 20µL

5.5

5.5.2 Multi-channel pipette . – capable of 20µL 5.5.3 Sterile pipette tips. – capable of 20µL 5.5.4 Stomacher® . – Seward or equivalent 5.5.5 Filter Stomacher® bags . - Seward or equivalent 5.5.6 Thermometer . – calibrated range to include 100 ± 1°C range 5.5.7 Incubators . – capable of maintaining 37 ± 1°C 5.5.8 Dry double block heater unit. – capable of maintaining 100 ± 1°C; OR a water bath capable of maintaining 100 ± 1°C 5.5.9 Freezer . – capable of maintaining -10 to -20°C, for storing the 3M Molecular Detection Chill Block Tray 5.5.10 Refrigerator . – capable of maintaining 2-8°C, for storing the 3M MDA 5.5.11 Computer . – compatible with the 3M Molecular Detection System (instrument)

Determination 5.6.1 Enrichment

5.6

5.6.1.1 Deli turkey - aseptically add the 125g test portion to 1125 mL Demi Fraser broth. 5.6.1.1.1 Homogenize thoroughly in a filter bag for 2 ± 0.2 min 5.6.1.1.2 Incubate 24-30 hr at 37 ± 1°C. 5.6.1.2 Raw chicken breast - aseptically add the 25g test portion to 475 mL Demi Fraser broth. 5.6.1.2.1 Homogenize thoroughly in a filter bag for 2 minutes ± 30 seconds 5.6.1.2.2 Incubate 28-32 hr at 37 ± 1°C. 5.6.2 Prepare the 3M Molecular Detection speed loader tray, chill block insert, heat block insert and instrument. 5.6.3 Equilibrate the LS tubes to room temperature (20-25 ºC) by setting LS tube overnight 16-18 hours. Or on the laboratory bench for at least 2 hours. 5.6.4 Invert the capped LS tubes to mix, up to 4 hours before use. 5.6.5 Transfer 20 µL enriched sample into a LS tube. AOAC Research I stitute Expert Review Panel Use Only

5/4/16