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Title: DEVELOPMENT OF AN ENZYME-LINKED IMMUNOSORBENT ASSAY FOR THE DETECTION OF ANTIBODY TO EPIZOOTIC HEMORRHAGIC DISEASE OF DEER VIRUS

Author
item Mecham, James
item JOCHIM, MICHAEL - VETERINARY DIAGNOSTIC TEC

Submitted to: Journal of Veterinary Diagnostic Investigation
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/16/1999
Publication Date: 3/20/2000
Citation: Mecham, J. O., Jochim, M. M. 2000. Development of an enzyme-linked immunosorbent assay for the detection of antibody to epizootic hemorrhagic disease of deer virus. Journal of Veterinary Diagnostic Investigation. 12:142-145.

Interpretive Summary: Epizootic hemorrhagic disease of deer virus (EHDV) often causes a fatal hemorrhagic disease of white-tailed deer, and can cause a bluetongue-like illness in cattle. Two serotypes, designated EHDV-1 (New Jersey strain) and EHDV-2 (Alberta strain), are enzootic in the United States. Because of their potential for causing disease in domestic livestock, it is important to have sensitive and specific diagnostic tools. Traditionally, the primary tests used for detection of antibodies to both bluetongue virus (BTV) and EHDV have been agar gel immunodiffusion (AGID) tests; however, serological cross-reactivity between these two groups of viruses can lead to false diagnoses. Since such false positives can create problems for importing and exporting cattle and cattle germplasm, it is important to have diagnostics capable of differentiating viruses in these two groups. Enzyme-linked immunosorbent assay (ELISA) procedures, that are highly sensitive and specific, have been developed for the detection of antibody to BTV. This paper describes the development of ELISA procedures based on both a competitive and a blocking format that can be used for EHDV diagnostics. The assays are extremely sensitive for the detection of antibodies to EHDV, and can distinguish animals infected with these viruses from those infected with BTV. In addition, the assays detect specific antibody in different species. These tests provide additional diagnostic tools that will be of value to both veterinarian diagnosticians and researchers.

Technical Abstract: An enzyme linked immunosorbent assay has been developed to detect antibodies to epizootic hemorrhagic disease of deer virus (EHDV). The assay incorporates a monoclonal antibody to EHDV serotype 2 (EHDV-2) that demonstrates specificity for the viral structural protein, VP7. The assay was evaluated with sequential sera collected from cattle experimentally infected with EHDV serotype 1 (EHDV-1) and EHDV-2, as well as the four serotypes of bluetongue virus (BTV), BTV-10, BTV-11, BTV-13 and BTV-17, that currently circulate in the U.S. A competitive and a blocking format as well as the use of antigen produced from both EHDV-1 and EHDV-2 infected cells were evaluated. The assay was able to detect specific antibody as early as 7 days after infection and could differentiate animals experimentally infected with EHDV from those experimentally infected with BTV. The diagnostic potential of this assay was demonstrated with field-collected serum samples from cattle, deer and buffalo.