ERP Micro December 2019

METHOD AUTHORS ORIGINAL VALIDATION: Ron Johnson and John Mills MODIFICATION DECEMBER 2018: Ronald Johnson, John Mills, Vikrant Dutta, Nikki Taylor, Deborah Briese

SUBMITTING COMPANY bioMérieux, Inc. 595 Anglum Road Hazelwood, MO 63042

KIT NAME(S) GENE-UP® Listeria monocytogenes 2 (LMO 2) Formerly known as GENE-UP® Listeria monocytogenes Detection System

CATALOG NUMBERS 423 107

INDEPENDENT LABORATORY Q Laboratories, inc. 1400 Harrison Ave Cincinnati, OH 45214 USA

AOAC EXPERTS AND PEER REVIEWERS Yi Chen 1, 4 , Michael Brodsky 2 , Wayne Ziemer 3 1 US FDA, CFSAN, College Park, MD, USA 2 Brodsky Consultants, Thornhill, ON, Canada 3 Consultant, Loganville, GA, USA 4 Modifications: October 2017, December 2018

APPLICABILITY OF METHOD Target organism – Listeria monocytogenes

REFERENCE METHODS United States Department of Agriculture Microbiological Laboratory Guidelines 8.09: Isolation and Identification of Listeria monocytogenes from Red Meat, Poultry, Egg Products, and Environmental Sponges . May 1 st , 2013. (Accessed September 2016) http://www.fsis.usda.gov/wps/wcm/connect/1710bee8-76b9-4e6c-92fc- dc290dbfa92/MLG-8.pdf?MOD=AJPERES (2) Food and Drug Administration Bacteriological Analytical Manual Chapter 10: Detection and Enumeration of Listeria monocytogenes in Foods . February, 2013. (Accessed September 2016) http://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm071400.htm (3)

Matrices – deli ham (25g, 125 g), deli roast beef (25 g), deli turkey (25 g), turkey hot dogs (25 g), smoked salmon (25 g), cooked shrimp (25 g), fresh spinach (25 g), mixed bagged salad (25 g), liquid whole egg (100 g), whey powder (375 g), vanilla ice cream (25 g), Mexican soft cheese (25 g, 125 g) Modification December 2018: stainless steel (sponge, 4 x 4 inch) Performance claims - The results of this study demonstrate that there are no significant differences between the GENE-UP method and the reference methods for the detection of Listeria monocytogenes .

ORIGINAL CERTIFICATION DATE December 23, 2016 METHOD MODIFICATION RECORD 1. October 2017 Level 2 2. December 2018 Level 2

CERTIFICATION RENEWAL RECORD Renewed annually through December 2019

SUMMARY OF MODIFICATION 1.

GENE-UP® software version 2.0 evaluation

2. Modifications include the following: (1) Manufacturing location change from Salt Lake City, Utah to Grenoble bioMérieux facility (2) a change from multi dose format to a unit dose format (3) Software updated to version 3 (4) PTM number change from 121602 to 121804. Under this AOAC® Performance Tested SM License Number, 121804 this method is distributed as: NONE

Under this AOAC® Performance Tested SM License Number, 121804 this method is distributed by: NONE

PRINCIPLE OF THE METHOD (1) The GENE-UP ™ Listeria monocytogenes (LMO) assay is used for the rapid and specific detection of Listeria monocytogenes in enriched food and environmental samples. The GENE-UP ™ Listeria monocytogenes assay is to be used with compatible PCR strip tubes in the GENE-UP ™ Thermocycler. Each reaction vial in the GENE-UP ™ Listeria monocytogenes assay contains all of the necessary components for PCR, including sample-specific primers and probes and an internal amplification control. The GENE-UP ™ Listeria monocytogenes assay detects fluorescence at several wavelengths (channels) to allow for multi-target detection in the same reaction vessel. The software automatically interprets the results for the internal amplification control and determines the sample result based on the outcome of the control. Both the assay for the sample and the internal amplification control utilize dual Fluorescence Resonance Energy Transfer (FRET) hybridization probes. The probes consist of two different short oligonucleotides that hybridize to an internal sequence of the amplified fragment during the annealing phase of the reaction cycle. FRET only occurs after the two probes come in close proximity from hybridizing to the template DNA. The resulting fluorescent signal from the FRET interaction, which forms a real time amplification curve, is how the amplified target is detected by the GENE-UP ™ Thermocycler. After the PCR cycling program finishes, the PCR product(s) are melted to determine the presence of the target DNA. The software uses both the real- time amplification curve and the melt peak to make a positive or negative call. [6]