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Journal of Clinical Microbiology, October 2004, p. 4641-4648, Vol. 42, No. 10
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.10.4641-4648.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Performance of Immunoglobulin G (IgG) and IgM Enzyme-Linked Immunosorbent Assays Using a West Nile Virus Recombinant Antigen (preM/E) for Detection of West Nile Virus- and Other Flavivirus-Specific Antibodies

Wayne R. Hogrefe,* Ronald Moore, Mary Lape-Nixon, Michael Wagner, and Harry E. Prince

Focus Technologies, Cypress, California

Received 11 February 2004/ Returned for modification 26 March 2004/ Accepted 23 June 2004

Focus Technologies developed an indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and a mu-capture IgM ELISA for the detection of West Nile virus (WNV)-specific antibodies based on a WNV preM/E protein recombinant antigen. Normal and disease state serum panels were used to assess the performance characteristics of the two WNV ELISA kits. Totals of 807 and 1,423 sera were used to assess the IgG ELISA and IgM ELISA kits, respectively. The Focus Technologies IgG ELISA had a sensitivity of 97.6% and a specificity of 92.1% (excluding non-WNV flavivirus sera). The comparative method for WNV IgG may lack sensitivity in detecting IgG in early WNV infection, so the specificity of the Focus IgG ELISA may be higher than 92.1%. When sera from patients either infected with or vaccinated against other flaviviruses were tested on the WNV IgG assay, 35% of the sera reacted as positive for WNV IgG. Yellow fever and Japanese encephalitis vaccinees were less reactive in the IgG ELISA than St. Louis and dengue fever patients. The Focus Technologies IgM ELISA had a sensitivity and a specificity of 99.3% (excluding the non-WNV flavivirus sera). The overall cross-reactivity for the IgM ELISA to flavivirus sera was 12%, with 31% of St. Louis encephalitis patients found to be WNV IgM positive and no yellow fever vaccinees found to be WNV IgM positive. In a selected population of 706 sera, 15 false-positive WNV IgM sera were identified. The use of a background subtraction method for the IgM ELISA eliminated all 15 false-positive results, giving a specificity of 100% for the Focus IgM ELISA.

* Corresponding author. Mailing address: Focus Technologies, 5785 Corporate Ave., Cypress, CA 90630. Phone: (714) 220-1900. Fax: (714) 821-3364. E-mail: whogrefe{at}focustechnologies.com.

Journal of Clinical Microbiology, October 2004, p. 4641-4648, Vol. 42, No. 10
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.10.4641-4648.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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