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Clinical and Diagnostic Laboratory Immunology, July 2004, p. 651-657, Vol. 11, No. 4
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.4.651-657.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Enzyme-Linked Immunosorbent Assays Using Recombinant Envelope Protein Expressed in COS-1 and Drosophila S2 Cells for Detection of West Nile Virus Immunoglobulin M in Serum or Cerebrospinal Fluid

A. Scott Muerhoff,1* George J. Dawson,1 Bruce Dille,1 Robin Gutierrez,1 Thomas P. Leary,1 Malini C. Gupta,1 Charles R. Kyrk,1 Hema Kapoor,2 Patricia Clark,2 Gerald Schochetman,1 and Suresh M. Desai1

Infectious Diseases Research and Development, Abbott Diagnostics, Abbott Laboratories, Abbott Park, Illinois,1 Bureau of Laboratories, Division of Infectious Diseases, Michigan Department of Community Health, Lansing, Michigan2

Received 6 February 2004/ Returned for modification 26 February 2004/ Accepted 16 March 2004

Humans infected with West Nile virus (WNV) develop immunoglobulin M (IgM) antibodies soon after infection. The microtiter-based assays for WNV IgM antibody detection used by most state public health and reference laboratories utilize WNV antigen isolated from infected Vero cells or recombinant envelope protein produced in COS-1 cells. Recombinant antigen produced in COS-1 cells was used to develop a WNV IgM capture enzyme immunoassay (EIA). A supplementary EIA using WNV envelope protein expressed in Drosophila melanogaster S2 cells was also developed. Both assays detected WNV IgM in the sera of experimentally infected rhesus monkeys within approximately 10 days postinfection. Human sera previously tested for WNV IgM at a state public health laboratory (SPHL) were evaluated using both EIAs. Among the sera from 20 individuals with laboratory-confirmed WNV infection (i.e., IgM-positive cerebrospinal fluid [CSF]) that were categorized as equivocal for WNV IgM at the SPHL, 19 were IgM positive and one was negative by the new EIAs. Of the 19 IgM-positive patients, 15 were diagnosed with meningitis or encephalitis; the IgM-negative patient was not diagnosed with neurological disease. There was 100% agreement between the EIAs for the detection of WNV IgM. CSF samples from 21 individuals tested equivocal for WNV IgM at the SPHL; all 21 were positive in both bead assays, and 16 of these patients were diagnosed with neurological disease. These findings demonstrate that the new EIAs accurately identify WNV infection in individuals with confirmed WNV encephalitis and that they exhibit enhanced sensitivity over that of the microtiter assay format.

* Corresponding author. Mailing address: Infectious Diseases Research and Development, Dept. 09NB, Bldg. AP20, Abbott Laboratories, 100 Abbott Park Rd., Abbott Park, IL 60064-6015. Phone: (847) 938-1077. Fax: (847) 938-6042. E-mail: scott.muerhoff{at}abbott.com.

Clinical and Diagnostic Laboratory Immunology, July 2004, p. 651-657, Vol. 11, No. 4
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.4.651-657.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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