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An enzyme immunoassay was developed for the detection of human immunoglobulin M (IgM) antibody to different flavivirus antigens. The IgM antibody of human sera was selectively bound to anti-IgM antibody-coated solid-phase plates. Flavivirus IgM antibodies were then detected by use of various enzyme-labeled antigens. The flavivirus antigens (dengue type 2 virus, West Nile virus, and tick-borne encephalitis virus) were produced in suckling mice. The antigens were labeled with horseradish peroxidase by adding the activated enzyme at alkaline pH to sucrose-acetone-treated antigens. Addition of unlabeled mouse brain suspension of uninfected animals to the diluted enzyme-labeled antigens effectively reduced nonspecific binding to the solid phase. In patients with acute flavivirus infections, viral IgM antibody could be demonstrated with high sensitivity. Furthermore, the enzyme-labeled antigen-IgM test showed greater specificity than the hemagglutination inhibition test.

Detection of specific immunoglobulin M antibody to different flaviviruses by use of enzyme-labeled antigens