AJTMH Transactions of the Royal Society of Tropical Medicine and Hygiene
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Am. J. Trop. Med. Hyg., 77(1), 2007, pp. 159-163
Copyright © 2007 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT


Flavivirus Serology by Western Blot Analysis

Leopoldo F. Oceguera, III, Peter J. Patiris*, Robert E. Chiles, Michael P. Busch, Leslie H. Tobler, AND Carl V. Hanson
California Department of Health Services, Viral and Rickettsial Disease Laboratory, Richmond, California; Blood Systems Research Institute, University of California, San Francisco, California

 

ABSTRACT

The spread of West Nile virus (WNV) across the United States into areas with endemic flavivirus activity has complicated serologic surveillance of seasonal virus activity and diagnosis of infected individuals. Here we describe preliminary results from a comparison of serologic assays for flaviviruses: the reference plaque reduction neutralization test (PRNT), enzyme immunoassay (EIA), and a Western blot (WB) in which crude viral lysates were electrophoresed and blotted onto nitrocellulose. Human and chicken sera were tested and compared by each method against WNV and St. Louis encephalitis virus (SLEV). Antibody binding to three viral proteins determined WB interpretation: non-structural protein 1 (NS1), envelope (E), and pre-membrane (prM). WB results for a group of serially collected human plasma samples from WNV seroconverting blood donors were also correlated with transcription mediated amplification (TMA) and polymerase chain reaction (RT-PCR) results. Reactivity with NS1 appeared to be the most useful differentiating marker of WNV and SLEV infection in humans and chickens. Envelope protein was highly cross-reactive and, as indicated by additional results from dengue virus (DENV)-positive human sera, is perhaps useful serologically as a flavivirus group antigen.

Received January 5, 2007. Accepted for publication April 7, 2007.

Acknowledgments: We thank Rob Lanciotti at the CDC Fort Collins laboratory for performing the PRNT work.

Financial support: This study was supported by CDC Grant RO1-CI-000214 in support of WNV studies at Blood Systems Research Institute in San Francisco.

* Address correspondence to Peter Patiris, Department of Health Services, Viral and Rickettsial Disease Laboratory, Richmond, CA 94804. E-mail: ppatiris{at}dhs.ca.gov

Authors’ addresses: Leopoldo F. Oceguera III, Peter J. Patiris, Robert E. Chiles, and Carl V. Hanson, California Department of Health Services, Viral and Rickettsial Disease Laboratory, Richmond, CA 94804, Telephone: 510-307-8555, Fax: 510-307-8955. Michael P. Busch and Leslie H. Tobler, Blood Systems Research Institute, University of California, San Francisco, 270 Masonic Avenue, San Francisco, CA 94118.

Reprint requests: Peter J. Patiris, Department of Health Services, Viral and Rickettsial Disease Laboratory, Richmond, CA 94804. E-mail: ppatiris{at}dhs.ca.gov.




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P. J. Patiris, L. F. Oceguera III, G. W. Peck, R. E. Chiles, W. K. Reisen, and C. V. Hanson
Serologic Diagnosis of West Nile and St. Louis Encephalitis Virus Infections in Domestic Chickens
Am J Trop Med Hyg, March 1, 2008; 78(3): 434 - 441.
[Abstract] [Full Text] [PDF]


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Copyright © 2007 by the American Society of Tropical Medicine and Hygiene.