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Clinical and Diagnostic Laboratory Immunology, March 2003, p. 267-271, Vol. 10, No. 2
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.2.267-271.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Competitive-Inhibition Enzyme-Linked Immunosorbent Assay for Detection of Serum Antibodies to Caprine Arthritis-Encephalitis Virus: Diagnostic Tool for Successful Eradication

Lynn M. Herrmann,1* William P. Cheevers,2 Travis C. McGuire,2 D. Scott Adams,3 Melinda M. Hutton,2 William G. Gavin,4 and Donald P. Knowles1,2

Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630,1 Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164-7040,2 VMRD, Inc., Pullman, Washington 99163,3 GTC Biotherapeutics, Framingham, Massachusetts 01701-93224

Received 30 October 2002/ Returned for modification 7 January 2003/ Accepted 10 January 2003

A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) was evaluated for the detection of serum antibodies to the surface envelope (SU) of caprine arthritis-encephalitis virus (CAEV) in goats. This assay utilized 96-well microtiter plates containing CAEV-63 SU captured by monoclonal antibody (MAb) F7-299 and measured the competitive displacement of horseradish peroxidase-conjugated MAb GPB 74A binding by undiluted goat sera (F. Özyörük, W. P. Cheevers, G. A. Hullinger, T. C. McGuire, M. Hutton, and D. P. Knowles, Clin. Diagn. Lab. Immunol. 8:44-51, 2001). Two hundred serum samples from goats in the United States were used to determine the sensitivity and specificity of cELISA based on the immunoprecipitation (IP) of [35S]methionine-labeled viral antigens as a standard of comparison. A positive cELISA was defined as >33.2% inhibition of MAb 74A binding based on 2 standard deviations above the mean percent inhibition of 140 IP-negative serum samples. At this cutoff value, there were 0 of 60 false-negative sera (100% sensitivity) and 5 of 140 false-positive sera (96.4% specificity). Additional studies utilized IP-monitored cELISA to establish a CAEV-free herd of 1,640 dairy goats.

* Corresponding author. Mailing address: Animal Disease Research Unit, ARS-USDA, Pullman, WA 99164-6630. Phone: (509) 335-6068. Fax: (509) 335-8328. E-mail: lherrman{at}vetmed.wsu.edu.

Clinical and Diagnostic Laboratory Immunology, March 2003, p. 267-271, Vol. 10, No. 2
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.2.267-271.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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