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United States Department of Agriculture

Agricultural Research Service

Research Project: Technologies for Detecting and Determining the Bioavailability of Bacterial Toxins

Location: Foodborne Toxin Detection and Prevention

Title: Dynex: macrobead multiplex ELISA for STEC

Authors
item Bernard Iv, Clay
item Clotilde, Laurie
item Sequera, Dean
item Karmali, Anis
item Fusellier, Andrew
item Carter, John

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: October 15, 2011
Publication Date: N/A

Technical Abstract: Recently, the Centers for Disease Control and Prevention has identified that the top 6 non-O157 Shiga toxin-producing E. coli (STEC) are emerging pathogens presenting significant risks of disease. Conventional enzyme linked immunosorbent assay (ELISA) is a gold standard for testing for protein or antigen presence. However, a significant limitation of this assay resides in the fact that only one analyte can be assessed per microplate well. Thus, we investigated a new technology consisting of an automated chemiluminescent ELISA system based on macro beads, in which up to 10 analytes can be measured within a single well. In this study we performed a “proof of principle” by testing for three factors: Shiga toxins 1 and 2, as well as E. coli O157. We found this technology not only improves productivity, accuracy and repeatability by reducing the amount of human labor required, but reduces cost and sample size by performing all tests simultaneously on the same sample (multiplexing). A strong point of the platform is that a user can load any necessary sets/subsets of macro beads to perform required assays, with improved flexibility compared to manufactured-loaded arrays for multiplex analysis. Our data show that this system can be used to determine the pathogenicity (i.e., presence of Shiga toxins) and serotype (i.e., E. coli O157) of E. coli isolates.

Last Modified: 10/31/2014
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