Rapid O serogroup identification of the ten most clinically relevant STECs by Luminex microbead-based suspension array |
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Authors: | Lin Andrew Nguyen Lam Lee Teresa Clotilde Laurie M Kase Julie A Son Insook Carter J Mark Lauzon Carol R |
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Institution: | a U.S. Food and Drug Administration San Francisco District Laboratory, 1431 Harbor Bay Parkway, Alameda, CA 94502, United Statesb California State University, East Bay, Department of Biological Sciences, 25800 Carlos Bee Blvd., Hayward, CA 94542, United Statesc U.S. Department of Agriculture, Agricultural Research Services, 800 Buchanan St., Albany, CA 94710, United Statesd U.S. Food and Drug Administration, Center for Food Safety and Nutrition, 5100 Paint Branch Parkway, College Park, MD 20740, United States |
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Abstract: | Identification and serotyping of Shiga toxin-producing Escherichia coli during foodborne outbreaks can aid in matching clinical, food, and environmental isolates when trying to identify the source of illness and ultimately food contamination. Herein we describe a Luminex microbead-based suspension array to identify the O serogroup of the ten most clinically relevant STECs: O26, O45, O91, O103, O111, O113, O121, O128, O145, and O157. The use of PCR followed by Luminex xMAP® technology enables the detection of multiple analytes in a single multiplex reaction with high throughput capabilities. One hundred and fourteen STEC isolates were correctly identified with no false positives among forty-six other organisms using this assay. Assay performance was tested in multiple laboratories using a panel of eleven different STEC serogroups on the Bio-Plex 200 and MAGPIX instruments. The STEC microbead-based suspension array can be performed in a 96-well plate format for high throughput screening in less than 4 h. Furthermore, it is expandable, allowing for the addition of O serogroups should the need arise. |
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Keywords: | STEC O serogroup identification Multi-plex PCR Luminex |
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