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Journal of Clinical Microbiology, February 2004, p. 518-523, Vol. 42, No. 2
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.2.518-523.2004

Antigen Capture Competitive Enzyme-Linked Immunosorbent Assays Using Baculovirus-Expressed Antigens for Diagnosis of Bluetongue Virus and Epizootic Hemorrhagic Disease Virus

J. O. Mecham^* and W. C. Wilson

Arthropod-Borne Animal Diseases Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Laramie, Wyoming

Received 7 August 2003/ Returned for modification 28 August 2003/ Accepted 4 November 2003

Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) are orbiviruses that infect both livestock and wild ruminants. Antigenic cross-reactivity between BTV and EHDV often results in serologic misdiagnosis. Competitive enzyme-linked immunosorbent assays (c-ELISAs) show increased sensitivity and specificity for the identification of these viral diseases; however, the preparation of cell culture-derived viral antigen for these tests is laborious and variable from batch to batch, and the resulting antigen may be infectious. To overcome these problems, the genes coding for a structural protein, VP7, of BTV and EHDV were cloned into baculovirus and the recombinant proteins were expressed in Sf9 cultured insect cells. Recombinant viral proteins released into the baculovirus-infected Sf9 cell culture supernatant were used in antigen capture c-ELISAs (Ag Cap c-ELISA) tests that specifically detected antibody in the serum of cattle experimentally infected with BTV and EHDV. The diagnostic utility of the Ag Cap c-ELISA was demonstrated by comparison with a commercial c-ELISA. The Ag Cap c-ELISA offers the advantages of using an easily produced, easily standardized, noninfectious antigen that does not require further purification or concentration.

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* Corresponding author. Mailing address: Arthropod-Borne Animal Diseases Research Laboratory, USDA, ARS, College of Agriculture, Dept. 3354, 1000 E. University Ave., Laramie, WY 82071. Phone: (307) 766-3620. Fax: (307) 766-3500. E-mail: jmecham{at}uwyo.edu.

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Journal of Clinical Microbiology, February 2004, p. 518-523, Vol. 42, No. 2
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.2.518-523.2004

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