ERP Micro December 2019

bioMérieux, Inc. GENEUP ® Listeria monocytogenes 2 (LMO 2) AOAC ® Certification Number 121804

DISCUSSION OF THE VALIDATION STUDY (1) The GENE-UP Listeria monocytogenes Assay successfully detected Listeria monocytogenes from all of the food matrices analyzed. Using POD analysis, no statistically significant differences were observed between the number of positive samples detected by the candidate method and the reference methods for all matrices tested. There was one matrix, deli ham (25 g), that resulted in 13 presumptive positive and only 8 test portions confirmed positive using the traditional confirmation method (MOX). When the ALOA plate was used, all 13 test portions confirmed positive. Additionally, there were no unexpected results in the lot-to-lot/stability and robustness studies and the inclusivity/exclusivity results showed perfect agreement. When comparing the alternative confirmation protocol (streak primary enrichment to ALOA followed by identification) to the traditional confirmation protocol (following complete reference method protocol, starting with secondary enrichment and/or agar plates), the alternative confirmation protocol resulted in 151/152 confirmed samples and the traditional confirmation protocol resulted in 143/152 confirmed samples. The GENE-UP monocytogenes species method is quick and simple to perform, providing results in less than 1.5 hours post incubation of the selective enrichment for 30 sample replicates. The GENE-UP software is user friendly with the ability to track lot information and sample identification quickly and with ease. The GENE-UP software and instrument also offer the ability to utilize an open platform and set up unique runs before a run is completed. The software also provides the user to analyze each individual CT curve and melting curve as well to help aid in problem solving any issues within an individual reaction. The small footprint is also desirable when working in laboratories with a limited amount of space, or a small PCR amplification room.

Table 1. GENE-UP Listeria monocytogenes Inclusivity Results (1) Reference a Strains

Origin

Assay

Result

Listeria monocytogenes 1/2a

Beacon

LMO

Positive

1402273

Listeria monocytogenes 1/2a

Ham

LMO

Positive

1402272

Listeria monocytogenes 1/2a

Cheese (Pasteurized)

LMO

Positive

904046

Listeria monocytogenes 1/2a

Smoked Salmon

LMO

Positive

1402270

Listeria monocytogenes 1/2a

Kidney (Pork)

LMO

Positive

1402283

Listeria monocytogenes 1/2a

Environmental Sample

LMO

Positive

1402282

Listeria monocytogenes 1/2b

Raw Milk Cheese

LMO

Positive

906247

Listeria monocytogenes 1/2b

Cheese (Pasteurized)

LMO

Positive

1402274

Listeria monocytogenes 1/2c

Raw Milk Cheese

LMO

Positive

904041

Listeria monocytogenes 1/2c

Raw Milk Cheese

LMO

Positive

904043

Listeria monocytogenes 1/2c

Raw Milk Cheese

LMO

Positive

904045

Listeria monocytogenes 1/2c

Fish Fillet

LMO

Positive

904049

Listeria monocytogenes 1/2c

Environmental Sample

LMO

Positive

1402256

Listeria monocytogenes 3a

Collection (ATCC b 51782)

LMO

Positive

904062

Listeria monocytogenes 3b

Sausage

LMO

Positive

1402266

Listeria monocytogenes 3c

Liver (veal)

LMO

Positive

1402255

Listeria monocytogenes 4a

Fish Fillet

LMO

Positive

1402254

Listeria monocytogenes 4b

Raw Milk Cheese

LMO

Positive

904042

Listeria monocytogenes 4b

Shellfish

LMO

Positive

1402251

Listeria monocytogenes 4d

Collection (ATCC 19117)

LMO

Positive

9409081

Listeria monocytogenes 4e

Collection (ATCC 19118)

LMO

Positive

904067

Listeria monocytogenes 4e

Salt meat

LMO

Positive

8309033

Listeria monocytogenes

Smoked salmon

LMO

Positive

9408051

Listeria monocytogenes

Raw Milk

LMO

Positive

9501016

Listeria monocytogenes

Salt meat

LMO

Positive

9311018

Listeria monocytogenes

LMO

Positive

0904025

Ready cooked dish

Listeria monocytogenes 1/2

Environmental sample

LMO

Positive

0010035

Listeria monocytogenes 1/2a

Raw beef meat

LMO

Positive

0010036

Listeria monocytogenes 1/2b

Smoked Salmon

LMO

Positive

8309023

Listeria monocytogenes 1/2c

Merguez sausage

LMO

Positive

0712183