Department of Veterinary Tropical Diseases, University of Pretoria, Onderstepoort 0110, South Africa.
J Virol Methods. 2011 Jun;174(1-2):60-4. doi: 10.1016/j.jviromet.2011.03.024. Epub 2011 Mar 31.
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.
本研究建立了一种基于多克隆抗体的、针对马脑炎病毒（EEV）的群特异性竞争 ELISA（C-ELISA）检测方法。该方法通过预滴定的 EEV 抗原，检测特定豚鼠抗血清与测试血清之间的竞争。C-ELISA 可检测到针对已知 7 种 EEV 血清型的抗体。针对其他虫媒病毒的参考抗血清与 EEV 抗原没有交叉反应。来自英国马的阴性血清用于建立阴性人群的基线。随后，根据病毒中和试验选择的南非马的阴性和阳性群体进行检测。通过双图接收者操作特性（TG-ROC）分析计算出最佳的测试参数，即灵敏度≈特异性=100%，其截断值为 29.5%抑制率。结果表明，所描述的 EEV C-ELISA 具有较高的敏感性、特异性和可靠性。与可用于检测非洲马瘟病毒（AHSV）的 ELISA 联合使用，可以实现 EEV 和 AHSV 的血清学鉴别诊断。
