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Journal of Clinical Microbiology, January 2004, p. 65-72, Vol. 42, No. 1
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.1.65-72.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Detection of Human Anti-Flavivirus Antibodies with a West Nile Virus Recombinant Antigen Microsphere Immunoassay

Susan J. Wong,1* Valerie L. Demarest,1 Rebekah H. Boyle,1 Tian Wang,2 Michel Ledizet,3 Kalipada Kar,3 Laura D. Kramer,1 Erol Fikrig,2 and Raymond A. Koski3

Wadsworth Center, New York State Department of Health, Albany, New York,1 Yale University School of Medicine,2 L2Diagnostics, New Haven, Connecticut3

Received 14 March 2003/ Returned for modification 21 June 2003/ Accepted 4 October 2003

We report a new, suspended-microsphere diagnostic test to detect antibodies to West Nile (WN) virus in human serum and cerebrospinal fluid (CSF). The microsphere immunofluorescence assay can be performed in less than 3 h on specimens of <=30 µl. A recombinant WN virus envelope (E) protein antigen is covalently coupled to fluorescent polystyrene microspheres. After incubation with diluted serum or CSF, antibodies bound to the E protein antigen are detected with fluorescently labeled anti-human immunoglobulin antibody and flow analysis in a dual-laser Luminex 100 instrument. Retrospective testing of 833 sera from New York patients with suspected viral encephalitis demonstrated concordance with results obtained with the traditional enzyme-linked immunosorbent assay for immunoglobulin G (IgG) antibodies to WN virus (kappa = 0.85). One hundred eighty-eight (22.4%) of the samples, which were collected from June to November 2002, tested positive for antibodies to WN virus in the microsphere assay. Specimens depleted of IgG with anti-IgG antibody were reassayed to measure anti-E protein IgM antibodies and to provide an indication of current or recent WN virus infection. The assay also detects antibodies to E proteins from related flaviviruses, including St. Louis encephalitis, Japanese encephalitis, and dengue viruses. The new microsphere immunoassay provides a sensitive and rapid alternative to traditional enzyme-linked immunosorbent assays that detect antibodies to flavivirus E proteins. This assay can aid physicians and public health workers in the management of outbreaks of WN virus and related flaviviruses.

* Corresponding author. Mailing address: Wadsworth Center, New York State Department of Health, P.O. Box 22002, Albany, NY 12201-2002. Phone: (518) 486-4396. Fax: (518) 473-6150. E-mail: sjw03{at}health.state.ny.us.

Journal of Clinical Microbiology, January 2004, p. 65-72, Vol. 42, No. 1
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.1.65-72.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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