Rapid Preclinical Detection of Sheeppox Virus by a Real-Time PCR Assay

Authors: BALINSKY, COREY - UNIVERSITY OF CONNECTICUT; DELHON, GUSTAVO - UNIVERSITY OF ILLINOIS; Smoliga, George; PRARAT, MELANIE - USDA, APHIS, PIADC; FRENCH, RICHARD - UNIVERSITY OF CONNECTICUT; GEARY, STEVEN - UNIVERSITY OF CONNECTICUT; ROCK, DANIEL - UNIVERSITY OF ILLINOIS; Rodriguez, Luis

Submitted to: Journal of Clinical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/11/2007
Publication Date: 2/1/2008
Citation: Balinsky, C.A., Delhon, G., Smoliga, G.R., Prarat, M., French, R.A., Geary, S.J., Rock, D.L., Rodriguez, L.L. 2008. Rapid Preclinical Detection of Sheeppox Virus by a Real-Time PCR Assay. Journal of Clinical Microbiology. 46(2):438-442.

Interpretive Summary: Sheeppox virus (SPV) is a member of the Capripox (CaPV) genus of the Poxviridae family which also includes Goatpox (GTPV) and Lumpy skin disease viruses (LSDV). These reportable disease agents are foreign to the United Stated and important pathogens particularly in Africa, the Middle East and parts of Asia. This manuscript describes the first real-time PCR for CaPV that detects all three foreign animal disease agents. This assay can detect SPPV in buffy coats, nasal swabs, oral swabs, scabs and skin lesions as well as in lung and lymph nodes collected at necropsy. This single tube diagnostic assay can be performed in 2 hours or less and can detect viral DNA in preclinical, clinical and post mortem samples. This work is part of an effort to develop rapid diagnostic tests for major foreign animal diseases including foot-and-mouth disease, African swine fever, classical swine fever, rinderpest, vesicular stomatitis and contagious bovine pleuroneumonia.

Technical Abstract: Sheeppox virus (SPPV) is a member of the Capripoxvirus (CaPV) genus of the Poxviridae family. Members of this genus, which also includes goatpox (GTPV) and lumpy skin disease (LSDV) viruses, cause economically significant disease in sheep, goats and cattle. A rapid diagnostic assay for CaPV would be useful for disease surveillance as well as for detection of CaPV in clinical samples and outbreak management. Here we describe a fluorogenic probe hydrolysis (TaqMan) PCR assay designed for rapid detection of CaPV and tested on sheep experimentally infected with a virulent strain of SPPV. This assay can detect SPPV in buffy coats, nasal swabs, oral swabs, scabs and skin lesions as well as in lung and lymph nodes collected at necropsy. This single tube diagnostic assay can be performed in 2 hours or less and can detect viral DNA in preclinical, clinical and post mortem samples.