The use of chicken IgY in a double antibody sandwich ELISA for detecting African horsesickness virus

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Authors

Du Plessis, D.H.
Van Wyngaardt, Wouter
Romito, M.
Du Plessis, M.

Journal Title

Journal ISSN

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Publisher

Pretoria : Agricultural Research Council, Onderstepoort Veterinary Institute

Abstract

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 lgY directed against AHSV (serotype 3) was used as the secondary antibody. Since lgY and mouse lgG 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.

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The articles have been scanned in colour with a HP Scanjet 5590; 600dpi. Adobe Acrobat v.9 was used to OCR the text and also for the merging and conversion to the final presentation PDF-format.

Keywords

African horse sickness, Antibody sandwich, Chickens, Egg-yolk, Orbiviruses, AHS

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Citation

Du Plessis, DH, Van Wyngaardt, W, Romito, M, Du Plessis, M & Maree, S 1999, 'The use of chicken IgY in a double antibody sandwich ELISA for detecting African horsesickness virus’. Onderstepoort Journal of Veterinary Research, vol. 66, no. 1, pp. 25-28.