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Cloning and expression of Rift Valley fever virus nucleocapsid (N) protein and evaluation of a N-protein based indirect ELISA for the detection of specific IgG and IgM antibodies in domestic ruminants

Author:
Fafetine, J.M., Tijhaar, E., Paweska, J.T., Neves, L.C.B.G., Hendriks, J., Swanepoel, R., Coetzer, J.A.W., Egberink, H.F., Rutten, V.P.M.G.
Source:
Veterinary microbiology 2007 v.121 no.1-2 pp. 29-38
ISSN:
0378-1135
Subject:
recombinant antigens, cattle diseases, disease detection, immunoglobulin G, immunoglobulin M, protein synthesis, serodiagnosis, sheep, antibody detection, cattle, goats, Rift Valley fever, Rift Valley fever phlebovirus, enzyme-linked immunosorbent assay, sheep diseases, molecular cloning, nucleocapsid proteins
Abstract:
Serodiagnosis of Rift Valley fever (RVF) currently relies on the use of live or inactivated whole virus as antigens. The recombinant nucleocapsid (N) protein of RVF virus was tested for diagnostic applicability in an indirect enzyme-linked immunosorbent assay (I-ELISA), using sera from experimentally infected sheep (n = 128), vaccinated sheep (n = 240), and field-collected sera from sheep (n = 251), goats (n = 362) and cattle (n = 100). The N-protein based I-ELISA performed at least as good as VN and HI tests. In goat the diagnostic sensitivity (D-Sn) and specificity (D-Sp) of the I-ELISA was 100% when using the anti-species IgG conjugate. Using protein G as a detection system, the D-Sn and D-Sp in goats were 99.4% and 99.5%, in sheep field sera both 100%, in cattle 100% and 98.3%, respectively. The I-ELISA based on recombinant N-protein has the potential to complement the traditional assays for serodiagnosis of RVF. Advantages of the N-protein are its safety, stability and cost-effectiveness in use and production.
Agid:
703833