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Clinical and Vaccine Immunology, May 2009, p. 749-755, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00393-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Evaluation of an Epitope-Blocking Enzyme-Linked Immunosorbent Assay for the Diagnosis of West Nile Virus Infections in Humans{triangledown}

M. A. Loroño-Pino,1,2 J. A. Farfan-Ale,2 B. J. Blitvich,1,5 J. L. Beebe,3,6 R. G. Jarman,4 and B. J. Beaty1*

Department of Microbiology, Immunology and Pathology, 1690 Campus Delivery, Colorado State University, Fort Collins, Colorado 80523,1 Laboratorio de Arbovirología, Centro de Investigaciones Regionales Dr. Hideyo Noguchi, Universidad Autónoma de Yucatán, Av. Itzáes No. 490 x 59, Centro, Mérida, Yucatán, México CP 97000,2 Colorado Department of Public Health and Environment, 8100 Lowry Blvd., Denver, Colorado 80230,3 Department of Virology, United States Army Medical Component, Armed Forces Research Institute of Medical Sciences, 315/6 Rajvithi Road, Bangkok, Thailand 10400,4 Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, 2116 Veterinary Medicine Building, Ames, Iowa 50011-1250,5 Public Health Laboratory, San Luis Obispo County Health Agency, 2191 Johnson Avenue, P.O. Box 1489, San Luis Obispo, California 934066

Received 24 October 2008/ Returned for modification 8 December 2008/ Accepted 12 March 2009

An epitope-blocking enzyme-linked immunosorbent assay (b-ELISA) was evaluated for the diagnosis of West Nile virus (WNV) infections in humans. Sera from patients diagnosed with WNV infections from an outbreak in 2003 in Colorado and from patients diagnosed with dengue virus infections from Mexico and Thailand were tested with the b-ELISA. The b-ELISAs were performed using the WNV-specific monoclonal antibody (MAb) 3.1112G and the flavivirus-specific MAb 6B6C-1. Although the WNV-specific b-ELISA was effective in diagnosing WNV infections in humans from Colorado, it was not efficacious for diagnosing WNV infections in serum specimens from Mexico and Thailand. In serum specimens from patients from Colorado, the WNV b-ELISA and the WNV plaque reduction neutralization test showed an overall agreement of 91%. The sensitivity and specificity of the WNV b-ELISA were 89% and 92%, respectively, with a false-positive rate of 5%, based on receiver operating characteristic analysis. In contrast, false-positive rate results in specimens from the countries of Mexico and Thailand, where flaviviruses are endemic, were 79% and 80%, presumably due to the presence of antibodies resulting from previous dengue virus infections in Mexico and/or Japanese encephalitis virus infections or vaccination in Thailand. Thus, in regions where people have experienced previous or multiple flavivirus infections, the use of the b-ELISA for WNV diagnosis is contraindicated.

* Corresponding author. Mailing address: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523-1690. Phone: (970) 491-2988. Fax: (970) 491-8707. E-mail: bbeaty{at}colostate.edu

{triangledown} Published ahead of print on 25 March 2009.

Clinical and Vaccine Immunology, May 2009, p. 749-755, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00393-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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