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P3-205 An Independent Evaluation of the GENE-UP® EHEC Detection Method for the Detection of Non-O157 Shiga-toxin Producing Escherichia coli (STEC) and Escherichia coli O157:H7 in Foods

Wednesday, July 12, 2017
Exhibit Hall (Tampa Convention Center)
Patrick Bird , Q Laboratories, Inc. , Cincinnati , OH
Benjamin Bastin , Q Laboratories, Inc. , Cincinnati , OH
Joe Benzinger , Q Laboratories, Inc. , Cincinnati , OH
Erin Crowley , Q Laboratories, Inc. , Cincinnati , OH
James Agin , Q Laboratories, Inc. , Cincinnati , OH
David Goins , Q Laboratories, Inc. , Cincinnati , OH
Introduction:  Most Escherichia coli are harmless; however, some shiga toxigenic escherichia coli (STEC) produce toxins (stx1, stx2, and eae) that can cause serious illness. The GENE-UP® EHEC Detection Method, which consists of three GENE-UP® tests, STEC (EH1), the GENE-UP® E. coli O157:H7 (ECO), and the GENE-UP® STEC-Top 6 (EH2), is designed to detect the virulence genes, and identify the top seven STEC strains. The assays utilize proprietary PCR technology to detect fluorescence at several wavelengths to allow for multi-target detection in the same reaction vessel. The resulting signal forms a real-time amplification curve after the PCR cycling program is completed; the PCR products are melted to determine the presence of the target DNA. The software uses both the amplification curve and the melt peak to make a positive or negative interpretation.

Purpose:   This study was conducted to independently evaluate an alternative rapid method to the FDA/BAM Chapter 4A method for spinach (200g), the USDA/FSIS MLG 5.09 method for raw beef trim (375 g), and the USDA/FSIS MLG 5B.05 method for raw beef trim (375 g) and raw ground beef (25g and 375g) as part of the AOAC RI™ PTM validation process.

Methods:  Using 30 unpaired samples, 5 replicates were evaluated at a high inoculation level, 20 at a low inoculation level, and 5 at an uninoculated control level. Samples were analyzed by the alternative method, and confirmed according to the reference method and a proprietary alternative procedure.

Results: Results for the assay were analyzed by POD statistical analysis. No statistically significant differences were observed between the new method and the reference methods.

Significance:  The data from the study demonstrated that the GENE-UP® EHEC was a rapid and reliable method for the for the detection of non-O157 STEC Top 6 and E. coli O157:H7.

Back to: Poster Session 3 – Low-water Activity Foods, Packaging, Produce, Microbial Food Spoilage, Antimicrobials, Laboratory and Detection Methods, Water
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