title: A group-specific, indirect sandwich ELISA for the detection of equine encephalosis virus antigen.
abstract: A polyclonal antibody-based, group-specific, indirect, sandwich ELISA (S-ELISA) for the detection of equine encephalosis virus (EEV) antigen was developed. Purified EEV particles were titrated in the S-ELISA and the limit of detection was determined to be approximately 9.0 ng of antigen/ml (0.45 ng/well). Positive S-ELISA reactions were recorded with seven serologically distinct EEV serotypes. No cross-reactions were recorded with other arboviruses including African horse sickness virus (AHSV) serotypes 1-9, bluetongue serotypes 1-24, epizootic haemorrhagic disease serotypes 1-8 and isolate 318, and selected isolates of Palyam, Eubenangee, Corriparta, Warrego, Akabane and bovine ephemeral fever viruses. The assay proved to be sensitive and specific for the rapid detection of EEV in cell cultures and in homogenated suckling mouse brain (MB). The data generated in this study suggest that the ELISA will be valuable for epidemiological studies of EE and will assist in making a reliable differential diagnosis between EEV and AHSV infections.
authors.affiliation: Department of Veterinary Tropical Diseases, Faculty of Veterinary Sciences, Private Bag X04, University of Pretoria, Onderstepoort 0110, South Africa. jcrafford@op.up.ac.za
authors.forename: J E
authors.initials: JE
authors.lastname: Crafford
authors.forename: A J
authors.initials: AJ
authors.lastname: Guthrie
authors.forename: M
authors.initials: M
authors.lastname: van Vuuren
authors.forename: P P C
authors.initials: PP
authors.lastname: Mertens
authors.forename: J N
authors.initials: JN
authors.lastname: Burroughs
authors.forename: P G
authors.initials: PG
authors.lastname: Howell
authors.forename: C
authors.initials: C
authors.lastname: Hamblin
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006168:Guinea Pigs; D006734:Horse Diseases / Q000097:blood / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D006736:Horses; D051379:Mice; D011817:Rabbits; D012087:Reoviridae / Q000276:immunology* / Q000302:isolation & purification; D012088:Reoviridae Infections / Q000097:blood / Q000175:diagnosis / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000956:Antigens, Viral