HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Monath, T. P. Articles by Nystrom, R. R. Search for Related Content PubMed PubMed Citation Articles by Monath, T. P. Articles by Nystrom, R. R.

Detection of Yellow Fever Virus in Serum by Enzyme Immunoassay^*

Yellow fever (YF) virus is present in patient's blood during the acute phase of illness. Virus isolation and identification provide a potential method of early diagnosis, but available techniques are slow and require specialized materials and equipment. An alternative approach is direct detection of YF antigen in serum by means of an enzyme-linked immunosorbent assay (ELISA). An antigen-capture ELISA was developed, which used anti-YF antibodies, immobilized on a solid phase (polystyrene plates), to capture YF virus from serum samples. After addition of the virus-containing sample, anti-YF detecting antibody conjugated to alkaline phosphatase was added to detect viral antigen. Trials with various capture and detecting antibodies in systems employing purified YF 17D virus, led to the selection of: 1) two capture antibodies (pooled human serum containing high titer YF IgM antibodies and a type-specific YF monoclonal antibody), and 2) a detecting antibody conjugate consisting of monoclonal antibody broadly cross-reactive with all flaviviruses, purified by affinity chromatography, and conjugated to alkaline phosphatase. The limit of sensitivity in tests against purified YF 17D virus diluted in buffer or normal human serum was 10^3.0–10^3.6 PFU/0.05 ml or 0.007–0.029 µg viral protein/0.05 ml. Sera obtained at intervals from rhesus and cynomolgus monkeys after infection with a wild YF virus strain were tested. The limit of sensitivity of the assay applied to viremic monkey serum was similar (approximately 3.5 log₁₀PFU/0.05 ml). Monoclonal capture antibody was more potent than human IgM for antigen capture from viremic sera containing high virus titers, whereas IgM provided a more sensitive assay for sera with low titers. Use of monoclonal capture antibody provided a specific virus identification. The procedure offers promise for use as a diagnostic test in human cases.

Accepted for publication August 4, 1983.

^* Address reprint requests to: Thomas P. Monath, M.D., Division of Vector-Borne Viral Diseases, Centers for Disease Control, Post Office Box 2087, Fort Collins, Colorado 80522.