title: Competitive ELISA for serodiagnosis of bluetongue: evaluation of group-specific monoclonal antibodies and expressed VP7 antigen.
abstract: The performance of 2 competitive enzyme-linked immunosorbent assays (C-ELISA) was compared with the reference C-ELISA I for the detection of antibodies to bluetongue virus (BTV). One of the assays (C-ELISA II) used a group-specific monoclonal antibody (MAb) to BTV, obtained from the American Type Culture Collection (8A3B-6) and tissue culture (TC)-derived BTV antigen (Ag), and the other assay (C-ELISA III) used BTV core protein VP7 (expressed in yeast) and the reference MAb (Pirbright Laboratory, 3-17-A3). Test sera were obtained by sequential blood samples from 22 calves, each inoculated with a different serotype (T) of BTV (South African [SA] T-1-T-16 and T-18-T-20 and USA T-11, T-13, and T-17). Sera were also obtained from 4 calves and 4 sheep inoculated with USA BTV T-10 and from several groups of calves exposed to single or multiple doses of epizootic hemorrhagic disease virus (EHDV) T-1-T-4 grown in TC (BHK-21) or suckling mouse brain (SMB). A total of 618 bovine and ovine field sera collected from BT-free and BT-endemic areas were also tested. The C-ELISA III was more sensitive than the C-ELISA II in the detection of anti-BTV antibody in sera from cattle and sheep early after infection with BTV. Seroconversion was demonstrated by the 3 C-ELISAs in all animals inoculated with BTV by 20 days postinfection (DPI), except in calves that received SA T-3 or USA T-13, which became positive at 40 DPI.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Animal Diseases Research Institute, Agriculture Canada, Nepean, Ontario.
authors.forename: A
authors.initials: A
authors.lastname: Afshar
authors.forename: B T
authors.initials: BT
authors.lastname: Eaton
authors.forename: P F
authors.initials: PF
authors.lastname: Wright
authors.forename: J E
authors.initials: JE
authors.lastname: Pearson
authors.forename: J
authors.initials: J
authors.lastname: Anderson
authors.forename: M
authors.initials: M
authors.lastname: Jeggo
authors.forename: H C
authors.initials: HC
authors.lastname: Trotter
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000097:blood*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology; D001667:Binding, Competitive; D001819:Bluetongue / Q000175:diagnosis*; D001820:Bluetongue virus / Q000276:immunology*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005779:Immunodiffusion; D012680:Sensitivity and Specificity; D012756:Sheep; D014758:Viral Core Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D014758:Viral Core Proteins; C066862:VP7 protein, orbivirus