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United States Department of Agriculture

Agricultural Research Service

Research Project: DETECTION OF TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY AGENTS IN LIVESTOCK, WILDLIFE, AGRICULTURAL PRODUCTS, AND THE ENVIRONMENT Title: Rapid multiplex immunoassay to distinguish botulinum neurotoxin serotypes on a single lateral flow device(Abstract)

Authors
item Ching, Kathryn
item Lin, Alice
item McGarvey, Jeffery
item Stanker, Larry
item Hnasko, Robert

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: August 4, 2012
Publication Date: N/A

Technical Abstract: Clostridium botulinum produces seven antigenically distinct serotypes of botulinum neurotoxin (BoNT/A–G). The potency of these toxins result in a high mortality rate with BoNT/A and /B accounting for most of the naturally occurring outbreaks. The ease of BoNT production and their potential use as biological weapons necessitates rapid and reliable BoNT detection by emergency personnel. The current standard for BoNT detection is a mouse bioassay followed by serotype determination. Thus, a portable and rapid assay capable of sensitive and selective BoNT serotype detection is needed in the event of a foodborne outbreak or a bioterrorism event. We have developed monoclonal antibody pairs directed against BoNT/A,/B and /E serotypes and describe their use in two colorimetric multiplex lateral flow immunoassays for detection of these BoNTs. Serotype specific monoclonal antibodies were either conjugated to gold particles or distinctly colored latex beads to visibly resolve individual serotype capture lines. Both the gold and latex bead immunoassays demonstrated high specificity in distinguishing BoNT serotypes on a single immunochromatographic test strip. All BoNT serotype antibody pairs were capable of detecting purified toxin or toxin spike beverages with maximal detection for BoNT/A (~1ng/mL). The latex-based multiplex assay demonstrated higher sensitivity and has the advantage of visual serotype identification by color code reducing potential user interpretation error.

Last Modified: 10/24/2014