Location: Arthropod-borne Animal Diseases ResearchTitle: Rift Valley fever virus structural and non-structural proteins: Recombinant protein expression and immunoreactivity against antisera from sheep Author
|Mcvey, D Scott - Scott|
Submitted to: Vector-Borne and Zoonotic Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/1/2013
Publication Date: 9/1/2013
Publication URL: http://handle.nal.usda.gov/10113/62669
Citation: Faburay, B., Wilson, W.C., Mcvey, D.S., Drolet, B.S., Weingartl, H., Madden, D., Young, A., Ma, W., Richt, J. 2013. Rift Valley fever virus structural and non-structural proteins: Recombinant protein expression and immunoreactivity against antisera from sheep. Vector-Borne and Zoonotic Diseases. 13(9):619-29. DOI: 10.1089. Interpretive Summary: Rift Valley Fever (RVF) is an acute viral zoonosis endemic in Africa and Arabian Peninsula. The disease is caused by the mosquito-transmitted arbovirus, Rift Valley fever virus (RVFV) in the family Bunyaviridae. Domestic and wild animals show abortions (pregnant animals) and high mortality rates (young animals). Humans infected by RVFV develop influenza-like symptoms ranging from fever to fatal encephalitis and hemorrhages. In this study a panel of recombinant proteins were developed that could be valuable for the development of diagnostic methods that differentiate infected from vaccinated animals (DIVA).
Technical Abstract: The Rift Valley fever virus (RVFV) encodes structural proteins, nucleoprotein (N), N-terminus glycoprotein (Gn), C-terminus glycoprotein (Gc) and L protein, 78-kDa and non-structural proteins NSm and NSs. Using the baculovirus system we expressed the full-length coding sequence of N, NSs, NSm, Gc and the ectodomain of the coding sequence of the Gn glycoprotein (Gne) derived from the virulent strain of RVFV, ZH548. Western blot analysis using anti-His antibodies and monoclonal antibodies against Gne and N confirmed expression of the recombinant proteins and in vitro biochemical analysis showed that the two glycoproteins, Gne and Gc, were expressed in glycosylated form. Immunoreactivity profiles of the recombinant proteins in western blot and in indirect enzyme-linked immunosorbent assay against a panel of antisera obtained from vaccinated or wild type (RVFV) challenged sheep confirmed the results obtained with anti-His antibodies and demonstrated the suitability of the baculo-expressed antigens for diagnostic assays. In addition, these recombinant proteins could be valuable for the development of diagnostic methods that differentiate infected from vaccinated animals (DIVA).