The use of chicken IgY in a double antibody sandwich ELISA for detecting African horsesickness virus.
An indirect sandwich ELISA that can detect as little as 8 ng of African horsesickness virus (AHSV) was developed. Viral antigen was captured from suspension using an immobilized monoclonal antibody specific for an epitope on VP7, a protein that is a major constituent of the virus core. Egg-yolk derived chicken IgY directed against AHSV (serotype 3) was used as the secondary antibody. Since IgY and mouse IgG do not cross-react serologically, the secondary antibody was not labelled, but was instead detected with enzyme-coupled sheep antibodies directed against avian immunoglobulins. The assay recognized all nine AHSV serotypes, but not the Cascara isolate of equine encephalosis virus, a related orbivirus that also infects horses. In addition to being able to detect and quantify whole AHSV, the ELISA could show the presence of VP7 produced by recombinant baculoviruses.
Immunology Division, Onderstepoort Veterinary Institute, South Africa.
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Du Plessis
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Van Wyngaardt
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Romito
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Du Plessis
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Maree
D000355:African Horse Sickness / Q000175:diagnosis*; D000356:African Horse Sickness Virus / Q000302:isolation & purification*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000032:analysis; D002645:Chickens / Q000276:immunology*; D004530:Egg Yolk / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006736:Horses; D007136:Immunoglobulins / Q000276:immunology*; D051379:Mice; D014758:Viral Core Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; C016512:IgY; D007136:Immunoglobulins; D014758:Viral Core Proteins