title: 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.
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.
authors.affiliation: Veterinary Faculty, Eduardo Mondlane University, Maputo, Mozambique, C. Postal 257, Mozambique. jfafetine@yahoo.com
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Fafetine
authors.forename: Edwin
authors.initials: E
authors.lastname: Tijhaar
authors.forename: Janusz T
authors.initials: JT
authors.lastname: Paweska
authors.forename: Luís C B G
authors.initials: LC
authors.lastname: Neves
authors.forename: Judith
authors.initials: J
authors.lastname: Hendriks
authors.forename: Robert
authors.initials: R
authors.lastname: Swanepoel
authors.forename: J A W
authors.initials: JA
authors.lastname: Coetzer
authors.forename: Herman F
authors.initials: HF
authors.lastname: Egberink
authors.forename: Victor P M G
authors.initials: VP
authors.lastname: Rutten
mesh_terms: D000820:Animal Diseases / Q000175:diagnosis / Q000276:immunology / Q000821:virology*; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000918:Antibody Specificity; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D015511:Goat Diseases / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D006041:Goats; D006385:Hemagglutination Inhibition Tests / Q000662:veterinary; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D009500:Neutralization Tests / Q000662:veterinary; D019590:Nucleocapsid Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012295:Rift Valley Fever / Q000097:blood / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012296:Rift Valley fever virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis / Q000276:immunology / Q000821:virology
chemical_list: D000914:Antibodies, Viral; D004279:DNA, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins