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Journal of Veterinary Diagnostic Investigation Vol. 19 Issue 2, 180-184
Copyright © 2007 by the American Association of Veterinary Laboratory Diagnosticians
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Brief Communications

Toward a multiplexed serotyping immunoassay for foot-and-mouth disease virus

Julie Perkins, Alfonso Clavijo, Josue I. Ortiz, Timothy J. Salo, Hilary J. Holland, Benjamin J. Hindson and Mary T. McBride1

Correspondence: 1Corresponding Author: Mary T McBride, Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, CA, 94551

Initial results demonstrating the feasibility of a multiplexed liquid array immunoassay for foot-and-mouth disease viral antigen detection and simultaneous serotype differentiation are presented. Serotype-specific antibodies from rabbit and guinea pig hyperimmunesera were isolated and prepared for use in a multiplexed, bead-based assay. The performance of all of the available antibodies as both capture and detector reagents was evaluated in the multiplexed system to establish a combination exhibiting the highest homotypic responses and lowest heterotypic reactions. The multiplexed assay was evaluated against inactivated cell culture supernatant samples of the same subtype as the virus used to raise the capture and detector antibodies. Distinct serotype differentiation was observed, except in the case of serotype SAT1. Subsequently, cell culture supernatant samples from a larger pool of viral subtypes were analyzed. Distinct serotype differentiation was obtained when analyzing cell culture supernatant samples from viral serotypes C, Asia, and SAT3, irrespective of the subtype. However, limitations of the current antibody pairs were realized in some inconclusive results obtained when analyzing samples from a broader range of O, A, and SAT2 subtypes. The results obtained in this initial study will be used to further optimize the assay using polyvalent or monoclonal antibodies and move toward the analysis of clinical samples.

Key Words: Foot-and-mouth disease • immunoassay • multiplexed • serotyping






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