Key Laboratories of Grazing Animal Diseases and Animal Virology, Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu 730046, China.
Vet J. 2010 Aug;185(2):211-5. doi: 10.1016/j.tvjl.2009.06.002. Epub 2009 Jul 7.
An indirect ELISA for the serological detection of bovine ephemeral fever virus (BEFV) infection in cattle is described in which a glycosylated protein of approximately 25 kDa (including the G1 antigenic site of the virus glycoprotein) expressed in Pichia pastoris GS115 was used as the coating antigen. The optimal concentration of coated antigen was 0.3 microg/well at a serum dilution of 1:40. The optimal positive threshold value of the assay was 1.88, as derived from receiver operating characteristic curve analysis. The test had 100% sensitivity and 96.7% specificity when compared with a micro-neutralisation test using 336 positive and 180 negative sera to BEFV, respectively. The inter-assay and intra-assay coefficients of variation for 15 sera were both <5.8% and there was no evidence of cross-reactivity between the tested coating antigen and antibodies to related rabies virus. The ELISA is an inexpensive and rapid serological detection method that would be suitable for screening for BEFV infection on a large scale.
本文介绍了一种用于检测牛流行热病毒（BEFV）感染的间接 ELISA 方法，该方法使用毕赤酵母 GS115 表达的约 25kDa 的糖基化蛋白（包括病毒糖蛋白的 G1 抗原位点）作为包被抗原。在血清稀释度为 1:40 时，最佳包被抗原浓度为 0.3μg/孔。通过接收者操作特征曲线分析，该测定的最佳阳性阈值为 1.88。与使用 336 份阳性和 180 份阴性 BEFV 血清的微量中和试验相比，该试验的敏感性为 100%，特异性为 96.7%。对 15 份血清进行的组内和组间变异系数均<5.8%，并且测试的包被抗原与相关狂犬病病毒抗体之间没有交叉反应的证据。该 ELISA 是一种廉价且快速的血清学检测方法，非常适合大规模筛查 BEFV 感染。
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