A competitive ELISA for the detection of group-specific antibody to equine encephalosis virus

• Published in: Journal of virological methods 2011-06, Vol.174 (1-2), p.60-64
• Main Authors: Crafford, J.E., Guthrie, A.J., Van Vuuren, M., Mertens, P.P.C., Burroughs, J.N., Howell, P.G., Batten, C.A., Hamblin, C.
• Format: Article
• Language: English
• Subjects: African horse sickness; African horse sickness virus; Animals; Antibodies; Antibodies, Viral - blood; Antisera; Biological and medical sciences; Clinical Laboratory Techniques - methods; Competitive ELISA; EEV; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Equine encephalosis virus; Fundamental and applied biological sciences. Psychology; Horse Diseases - diagnosis; Horse Diseases - virology; Horses; Microbiology; Orbivirus - immunology; Orbivirus - isolation & purification; Reoviridae Infections - diagnosis; Reoviridae Infections - veterinary; Reoviridae Infections - virology; Sensitivity and Specificity; Serogroup-specific antibody; Serotypes; South Africa; Techniques used in virology; United Kingdom; Virology; Virology - methods
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2011.03.024

A polyclonal antibody-based, group-specific, competitive ELISA (C-ELISA) for the detection of antibodies to equine encephalosis virus (EEV) was developed. The assay measures the competition between a specific guinea pig antiserum and a test serum, for a pre-titrated EEV antigen. The C-ELISA detected antibodies to the seven known EEV serotypes. Reference antisera raised against other arboviruses did not cross react with EEV antigen. Negative sera from horses in the United Kingdom were used to establish the baseline for a negative population. Negative and positive populations of South African horses, selected on the basis of virus neutralisation were assayed subsequently. Optimal test parameters, where sensitivity≅specificity≅100%, were calculated by two-graph receiver operator characteristic (TG-ROC) analysis to be at a cut-off value of 29.5% inhibition. Results show the EEV C-ELISA described to be sensitive, specific and reliable. Used in conjunction with ELISAs available for African horse sickness virus (AHSV), differential serological diagnosis between EEV and AHSV can be achieved.