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

5.6.6 Transfer 20 µL Negative Control [NC], sterile enrichment medium (e.g. Demi-Fraser Broth) into a Lysis Solution [LS] tube after all enriched samples have been completed. Do not use water as a NC. 5.6.7 Place uncovered LS tubes in 3M Molecular detection Heat block, heat 15±1 min at 100±1°C. LS Solution will change from pink (cool) to yellow (hot). 5.6.8 Place LS tubes (without rack lid) in chill block insert (at ambient temperature) for 5-10 min. Lysis solution in LS tube will revert to pink color. 5.6.9 Transfer 20 µL sample lysate from the upper portion of fluid in the LS tube into regent tube. Mix gently by pipetting up and down 5 times. 5.6.10 Transfer 20 µL of NC lysate into a reagent tube. 5.6.11 Transfer 20 µL of NC lysate into a Reagent Control (RC) tube. 5.6.12 When all samples have been transferred, load capped tubes into speed loader tray and close lid. 5.6.13 Start assay. 5.6.14 Results and Interpretation - An algorithm interprets the light output curve resulting from the detection of the nucleic acid amplification. Results are analyzed automatically by the software and are color-coded based on the result. A Positive or Negative result is determined by analysis of a number of unique curve parameters. Presumptive positive results are reported in real-time while Negative and Inspect results will be displayed after the run is completed. Presumptive positive results should be confirmed using your preferred method or as specified by local regulations. NOTE: Even a negative sample will not give a zero reading as the 3M MDS and 3M MDA amplification reagents have a “background” relative light unit (RLU). In the rare event of any unusual light output, the algorithm labels this as “Inspect.” 3M recommends the user to repeat the assay for any Inspect samples. If the result continues to be Inspect, proceed to confirmation test using your preferred method or as specified by local regulations. AOAC Research Institute Expert Review Panel Use Only 5.6.16 Confirmation All positive 3M MDA 2 - Listeria monocytogenes results must be confirmed. Confirmation should be performed using the primary enrichment, carried through as described in the reference method starting with the secondary transfer and selective isolation for deli turkey and selective isolation for raw chicken breast. Confirmation should be initiated within 72 hrs. Isolate on a selective agar and identify 1 to 5 typical colonies according to the conventional tests described in the AOAC or USDA/FSIS MLG methods. 5.6.15

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