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Journal of Clinical Microbiology, September 2003, p. 4217-4223, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4217-4223.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Immunoassay Targeting Nonstructural Protein 5 To Differentiate West Nile Virus Infection from Dengue and St. Louis Encephalitis Virus Infections and from Flavivirus Vaccination

Susan J. Wong,1* Rebekah H. Boyle,1 Valerie L. Demarest,1 Anh N. Woodmansee,1 Laura D. Kramer,1 Hongmin Li,1 Michael Drebot,2 Raymond A. Koski,3 Erol Fikrig,4 Denise A. Martin,5 and Pei-Yong Shi1*

Wadsworth Center, New York State Department of Health, and Department of Biomedical Sciences, University at Albany, State University of New York, Albany, New York 12201,1 National Microbiology Laboratory, Health Canada, Winnipeg, Manitoba, Canada R3E 3R2,2 L Squared Diagnostics,3 Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520,4 Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Fort Collins, Colorado 805225

Received 14 May 2003/ Returned for modification 10 June 2003/ Accepted 23 June 2003

West Nile virus (WNV) is an emerging flavivirus that has caused frequent epidemics since 1996. Besides natural transmission by mosquitoes, WNV can also be transmitted through blood transfusion and organ transplantation, thus heightening the urgency of development of a specific and rapid serologic assay of WNV infection. The current immunoassays lack specificity because they are based on detection of antibodies against WNV structural proteins and immune responses to structural proteins among flaviviruses cross-react to each other. Here, we describe microsphere immunoassays that detect antibodies to nonstructural proteins 3 and 5 (NS3 and NS5). In contrast to immunoassays based on viral envelope and NS3 proteins, the NS5-based assay (i) reliably discriminates between WNV infections and dengue virus or St. Louis encephalitis virus infections, (ii) differentiates between flavivirus vaccination and natural WNV infection, and (iii) indicates recent infections. These unique features of the NS5-based immunoassay will be very useful for both clinical and veterinary diagnosis of WNV infection.

* Corresponding author. Mailing address: Wadsworth Center, New York State Department of Health, Albany, NY 12201. Phone for Susan J. Wong: (518) 486-4396. Fax: (518) 473-6150. E-mail: sjw03{at}health.state.ny.us. Phone for Pei-Yong Shi: (518) 473-7487. Fax: (518) 473-1326. E-mail: ship{at}wadsworth.org.

Journal of Clinical Microbiology, September 2003, p. 4217-4223, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4217-4223.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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