Microsoft Word - Draft OMB Meeting Agenda-April 11 2019

OMAMAN-29 D/ PTM Validation Report 111501 OMA ERP - June 2016 ERP Use Only

OMB Meeting 4-11-2019 Pre-read Materials

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Environmental samples

Sample collection devices can be a sponge hydrated with a neutralizing solution to inactivate the effects of the sanitizers. 3M recommends the use of a biocide-free cellulose sponge. Neutralizing solution can be Dey-Engley (D/E) Neutralizing Broth or Letheen Broth. It is recommended to

sanitize the area after sampling.

The recommended size of the sampling area for verifying the presence or absence of the pathogen on the surface is at least 100 cm 2 (10 cm x 10 cm or 4”x4”). When sampling with a sponge, cover the entire area going in two directions (left to right then up and down) or collect environmental samples following your current sampling protocol or according to the FDA BAM,

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USDA FSIS MLG or ISO 18593 [7] guidelines.

1. Allow the Demi-Fraser Broth enrichment medium (includes FAC) to equilibrate to ambient

laboratory temperature.

2. Aseptically combine the enrichment medium and sample according to Table 1. 3. Homogenize thoroughly by blending, stomaching, or hand mixing for 2 ±0.2 minutes. 4. Invert room temperature (20-25 o C) lysis tubes to mix. Proceed to next step within 4 hours. 20 µL aliquot of each enriched sample are is transferred to separate lysis tubes using a new pipette tip after each sample transfer. Place uncovered samples on a dry double block heater r for 15 ± 1 minutes at 100 ± 1 o C. Following the heat lysis transfer samples into the chill block insert and placed onto a sterilized lab bench and allow to cool at 18-28 °C for 5-10 Incubate at 37 ±1°C for 24-30 hours. 5. Add 20 µL of each lysed sample and control to separate reagent tubes, and mix by pipetting up and down five times. Analyze a matrix control tube with the samples for each matrix to verify that no interference with the assay was caused by the matrix. Use sterile Demi Fraser 6. Transfer a 20 µL aliquot to the NC and the Reagent Control (RC) tubes. Add a 20 µL aliquot of a randomly picked sample to the matrix control tube, mixed, and recapped. Using the 3M ™ software, follow prompts to identify samples and controls. Load all samples into the Speed Loader Tray (SLT), place into the Molecular Detection System, and initiate the 3M ™ MDA 2 AOAC Research In titute Expert Review Panel Use Only - Listeria assay. Results are obtained within 75 minutes. Interpretation and Test Result Report 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 Broth with FAC for the Negative Control (NC). minutes.

results will be displayed after the run is completed.

NOTE: Even a negative sample will not give a zero reading as the system and 3M Molecular Detection Assay 2 - Listeria amplification reagents have a “background” relative light unit

(RLU) reading.

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

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