T10-07 Development of Latex Agglutination Tests for Non-O157 Shiga Toxin-producing Escherichia coli O26, O45, O103, O111, O121 and O145

Wednesday, July 25, 2012: 10:30 AM
Ballroom E (Rhode Island Convention Center)
Marjorie Medina, U.S. Department of Agriculture-ARS-ERRC, Wyndmoor, PA
Weilin Shelver, U.S. Department of Agriculture-ARS-BRL, Fargo, ND
Pina Fratamico, U.S. Department of Agriculture-ARS-ERRC, Wyndmoor, PA
Laurie Fortis, U.S. Department of Agriculture-NIFA-IBCE, Washington DC, DC
Glenn Tillman, U.S. Department of Agriculture-FSIS-OPHS, Athens, GA
Neelam Narang, U.S. Department of Agriculture-FSIS, Athens, GA
William Cray Jr., U.S. Department of Agriculture-FSIS, Athens, GA
Emilio Esteban, U.S. Department of Agriculture-FSIS-OPHS-EALS, Athens, GA
Chitrita DebRoy, The Pennsylvania State University, University Park, PA
Introduction:   Shiga toxin-producing Escherichia coli (STEC) strains are important foodborne pathogens responsible for numerous outbreaks.  The top six non-O157 STECs (O26, O45, O103, O111, O121 and O145) are responsible for 70% of non-O157 human illness in the United States.

Purpose:   Simple and rapid methods to detect and identify these pathogens are needed to help confirm these serogroups.  Latex agglutination assays were developed for detection of the top six non-O157 STECs.

Methods:   Immunoglobulins (IgG) were affinity purified from rabbit antisera and covalently immobilized onto polystyrene latex particles.  The optimum conditions for the agglutination assay consisted of utilizing 20 µl of latex-IgG reagent containing 2.0–2.8 mg IgG in a 0.5% latex suspension.

Results:   Agglutination was observed instantly after mixing the colonies with the latex-IgG indicating positive reactions for the target strains. This method detected and helped confirm the target serogroups.  Over 100 target and non-target strains were tested in more than 3,000 test replicates.  The anti-O103 and anti-O145 latex reagents showed cross-reactions with O26 strains while anti-O26 cross-reacted with serogroup O103.  The latex-IgG reagents are stable for at least one year.

Significance:   The latex-IgG reagents can be used for identification of presumptive positive non-O157 STEC colonies. The method of preparation of the latex reagents can also be utilized to prepare these types of reagents for other STEC serogroups and other pathogens to ensure safe foods to consumers.