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Journal of Clinical Microbiology, March 2003, p. 1041-1047, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.1041-1047.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Epitope-Blocking Enzyme-Linked Immunosorbent Assays for the Detection of Serum Antibodies to West Nile Virus in Multiple Avian Species

Bradley J. Blitvich,1 Nicole L. Marlenee,1 Roy A. Hall,2 Charles H. Calisher,1 Richard A. Bowen,3 John T. Roehrig,4 Nicholas Komar,4 Stanley A. Langevin,4 and Barry J. Beaty1*

Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology ,1 Animal Reproduction and Biotechnology Laboratory, Equine Center, Colorado State University, Fort Collins, Colorado 80523,3 Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado 80522,4 Department of Microbiology and Parasitology, University of Queensland, St. Lucia, Queensland, Australia 40722

Received 24 July 2002/ Returned for modification 6 November 2002/ Accepted 22 November 2002

We report the development of epitope-blocking enzyme-linked immunosorbent assays (ELISAs) for the rapid detection of serum antibodies to West Nile virus (WNV) in taxonomically diverse North American avian species. A panel of flavivirus-specific monoclonal antibodies (MAbs) was tested in blocking assays with serum samples from WNV-infected chickens and crows. Selected MAbs were further tested against serum samples from birds that represented 16 species and 10 families. Serum samples were collected from birds infected with WNV or Saint Louis encephalitis virus (SLEV) and from noninfected control birds. Serum samples from SLEV-infected birds were included in these experiments because WNV and SLEV are closely related antigenically, are maintained in similar transmission cycles, and have overlapping geographic distributions. The ELISA that utilized MAb 3.1112G potentially discriminated between WNV and SLEV infections, as all serum samples from WNV-infected birds and none from SLEV-infected birds were positive in this assay. Assays with MAbs 2B2 and 6B6C-1 readily detected serum antibodies in all birds infected with WNV and SLEV, respectively, and in most birds infected with the other virus. Two other MAbs partially discriminated between infections with these two viruses. Serum samples from most WNV-infected birds but no SLEV-infected birds were positive with MAb 3.67G, while almost all serum samples from SLEV-infected birds but few from WNV-infected birds were positive with MAb 6B5A-5. The blocking assays reported here provide a rapid, reliable, and inexpensive diagnostic and surveillance technique to monitor WNV activity in multiple avian species.

* Corresponding author. Mailing address: Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523. Phone: (970) 491-2988. Fax: (970) 491-8323. E-mail: bbeaty{at}colostate.edu.

Journal of Clinical Microbiology, March 2003, p. 1041-1047, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.1041-1047.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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