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Clin. Vaccine Immunol. doi:10.1128/CVI.00004-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Enzyme-linked immunosorbent assays using novel Japanese encephalitis virus antigen improves the accuracy of clinical diagnosis in flavivirus infected patients

Graduate Institute of Veterinary Public Health, College of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan, R.O.C.; Arboviral Diseases Branch, Division of Vector-Borne Infectious Diseases, National Center for Zoonotic, Vector-Borne, and Enteric Diseases, Center for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Service, Fort Collins, Colorado, U. S. A.; College of Medicine, Tzu-Ch University, Hualien, Taiwan, R.O.C.

^* To whom correspondence should be addressed. Email: gxc7{at}cdc.gov.

	Abstract

The cross-reactive antibodies induced by flavivirus infections confound serodiagnosis and pathogenesis, especially in secondary infections caused by antigenically closely related, yet distinct flaviviruses. The envelope (E) glycoprotein fusion peptide contains immunodominant cross-reactive determinants. Using a recombinant Japanese encephalitis virus (JEV) premembrane and E expression plasmid producing JEV virus-like particles (VLPs), dramatic reductions in cross-reactivity were produced by the G106K-L107D (KD) double-mutant VLP against a panel of flavivirus murine monoclonal antibodies. Human serum panels from patients with recent flavivirus infections were analyzed to compare the accuracy of JEV wild-type (WT) and KD VLPs as serodiagnostic antigens in enzyme-linked immunosorbent assays. Statistical analysis demonstrated significant differences in assay performance for accurate determination of current JEV infections between WT and KD antigens by detecting IgM antibodies at serum dilution of 1:4000 (likelihood ratio = 2.74 [WT] and 22 [KD]). The application and continued development of cross-reactivity reduced antigens should improve both flavivirus serodiagnosis and estimates of disease burden.