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Title: EPIZOOTIC HEMORRHAGIC DISEASE: ANALYSIS OF TISSUES BY AMPLIFICATION AND IN SITU HYBRIDIZATION REVEALS WIDESPREAD ORBIVIRUS INFECTION AT LOW COPY NUMBERS

Author
item BRODIE, SCOTT - UNIV. OF WASHINGTON
item Mecham, James
item NORELIUS, S. - UNIV. OF WASHINGTON
item DIEM, K. - UNIV. OF WASHINGTON
item Wilson, William - Bill

Submitted to: Journal of Virology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/1/1998
Publication Date: N/A
Citation: Brodie, S.J., Mecham, J.O., Norelius, S.E., Diem, K., Wilson, W.C. 1998. Epizootic hemorrhagic disease: analysis of tissues by amplification and in situ hybridization reveals widespread orbivirus infection at low copy numbers. Journal of Virology. 72:3863-71.

Interpretive Summary: Epizootic hemorrhagic disease virus is the causative agent of disease in white-tailed deer and is known to infect domestic ruminants. This study was part of a disease outbreak study that developed new tools to investigate the disease process of this virus. These tools will be used to further evaluate the risk of this virus to domestic livestock.

Technical Abstract: A recent outbreak of hemorrhagic fever in wild ruminants in the northwest United States was characterized by rapid onset of fever, followed shortly thereafter by hemorrhage and death. As a result, a confirmed 1,000 white-tailed deer and pronghorn antelope died over the course of 3 months. Lesions were multisystemic and included severe edema, congestion, acute vascular necrosis, and hemorrhage. Animals that died with clinical signs and/or lesions consistent with hemorrhagic fever had antibody to epizootic hemorrhagic disease virus serotype 2 (EHDV-2) by radioimmune precipitation but the antibody was limited exclusively to class immunoglobulin M. These findings, indicative of acute infection, were corroborated by the observation that numerous deer were found dead; however, clinically affected deer were rarely seen during the outbreak. Furthermore, only in animals with hemorrhagic lesions was EHDV-2 isolated and/or erythrocyte-associated EHDV-2 RNA detected by serotype-specific reverse transcription (RT)- PCR. By using a novel RT in situ PCR assay, viral nucleic acid was localized to the cytoplasm of large numbers of tissue leukocytes and vascular endothelium in tissues with hemorrhage and to vessels, demonstrating acute intimal and medial necrosis. Because PCR amplification prior to in situ hybridization was essential for detecting EHDV, the virus copy number within individual cells was low, <20 virus copies. These findings suggest that massive covert infection characterized by rapid dissemination of virus facilitates the severe and lethal nature of this disease.