Microsoft Word - Draft OMB Meeting Agenda-April 11 2019

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OMAMAN-31 E: AOAC RI PTM Report 091501 ERP Use Only - March 2016

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leading to the release of contaminated product. Another option is to transfer 10 uL of the

NB enrichment in the LS tubes.

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,

USDA FSIS MLG or ISO 18593 guidelines.

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1. Pre-warm BPW ISO enrichment medium to 41.5 ±1°C depending upon matrices tested.

See Table 1.

2. Aseptically combine the enrichment medium and sample.

3. Homogenize thoroughly by blending, stomaching, or hand mixing for 2 ±0.2 minutes.

Incubate at 41.5 ±1°C for 18-24 hours .

Post-enrichment

1. Invert room temperature (20-25 o C) capped lysis tubes to mix. Proceed to next step within 4 hours. A 20 µL aliquot of each enriched sample is transferred to a separate lysis tubes using a new pipette tip after each sample transfer. Place uncovered samples in the 3M Molecular Detection Heat Block for 15 ± 1 minutes at 100 ± 1 o C. Following the heat lysis, place samples in the 3M Molecular Detection Chill Block Insert for at least 5 minutes and a maximum of 10 minutes. Alternatively, the samples can be placed at room temperature for 5 2. 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 a sample of each matrix to verify that no interference with the assay was caused by the matrix. Use sterile 3M BPW ISO for the Negative Control (NC). Transfer a 20 µL aliquot to the NC and the Reagent Control (RC) 3. Add a 20 µL aliquot of a randomly picked sample to the matrix control tube, mix, and recap. 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 - Salmonella assay. Results are obtained within 75 minutes. to 10 minutes. tubes. 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 NOTE: Even a negative sample will not give a zero reading as the system and 3M MDA2 – Salmonella 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 results will be displayed after the run is completed. AOAC Research Institute Expert Review Panel Use Only Interpretation and Test Result Report

04/05/2019