Enzyme-linked immunosorbent assay for detection of bluetongue virus antibodies.
The immune response to bluetongue virus in sheep and cattle was studied by applying a newly developed indirect enzyme-linked immunosorbent assay (ELISA). Purified virus obtained by sucrose gradient centrifugation was used at a concentration of 0.01 optical density units (formula: see text) to coat individual wells (200 microliter) of a microtitration plate. Dilution of antigen was performed in 0.05 M carbonate buffer, pH 9.6, and adsorption lasted for at least 16 hours at 4 C. Coated plates retained their activity for 10 weeks when stored at 4 C. Sera recovered from experimentally infected sheep and cattle were tested together with known negative sera. A good correlation between results was obtained with the modified complement-fixation test and the ELISA; however, the ELISA proved to be more sensitive. The group specificity of the ELISA was proven by testing various type-specific sheep and cattle immune sera. The ELISA has potential for the detection of group-specific antibodies to bluetongue virus infection.
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D001819:Bluetongue / Q000276:immunology; D001820:Bluetongue virus / Q000187:drug effects / Q000276:immunology*; D002254:Carbonates / Q000494:pharmacology; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D007124:Immunoenzyme Techniques; D012087:Reoviridae / Q000276:immunology*; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology
D000914:Antibodies, Viral; D002254:Carbonates