title: Rapid O serogroup identification of the ten most clinically relevant STECs by Luminex microbead-based suspension array.
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 4h. Furthermore, it is expandable, allowing for the addition of O serogroups should the need arise.
authors.affiliation: U.S. Food and Drug Administration San Francisco District Laboratory, Alameda, CA 94502, USA. andrew.lin@fda.hhs.gov
authors.forename: Andrew
authors.initials: A
authors.lastname: Lin
authors.forename: Lam
authors.initials: L
authors.lastname: Nguyen
authors.forename: Teresa
authors.initials: T
authors.lastname: Lee
authors.forename: Laurie M
authors.initials: LM
authors.lastname: Clotilde
authors.forename: Julie A
authors.initials: JA
authors.lastname: Kase
authors.forename: Insook
authors.initials: I
authors.lastname: Son
authors.forename: J Mark
authors.initials: JM
authors.lastname: Carter
authors.forename: Carol R
authors.initials: CR
authors.lastname: Lauzon
mesh_terms: D008863:Microspheres; D060885:Multiplex Polymerase Chain Reaction; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D012703:Serotyping / Q000295:instrumentation / Q000379:methods*; D054323:Shiga-Toxigenic Escherichia coli / Q000145:classification*; D059629:Signal-To-Noise Ratio