牛病毒性腹泻病毒非结构蛋白NS3的表达及ELISA检测方法的建立

Expression of Nonstructural Protein NS3 and Establishment of ELISA Method of Bovine Viral Diarrhea Virus

本研究旨在建立一种检测牛病毒性腹泻病毒(BVDV)抗体的间接ELISA方法。将BVDV的非结构蛋白NS3基因克隆到原核表达载体pET-32a中进行表达,将纯化后的蛋白作为包被抗原,优化ELISA条件,建立了BVDV抗体间接NS3-ELISA检测方法,并对该方法的特异性、敏感性和重复性进行检测,结果均较好。用所建立的NS3-ELISA方法检测从广西各牛场采集的475份牛血清样品,检出率为24.8%,与商品化试剂盒比较,符合率为97%。结果表明,本研究建立的NS3-ELISA方法简便、快捷,可大批量检测,适用于BVDV的诊断、抗体水平监测及流行病学调查。

An indirect enzyme-linked immunosorbent assay （ELISA） method was developed to detect antibody against bo- vine viral diarrhea virus （BVDV）. The nonstruetural protein NS3 gene of BVDV was subcloned into prokaryotic vector pET-32a to express. The recombinant protein NS3 of BVDV was used as the coating antigen for the ELISA method, and the reaction conditions were optimized. The specificity, sensitivity and reproducibility all showed good results. Furthermore, four hundred and seventy five bovine serum samples, which had been collected from several cattle farms of Guangxi, were detected by the NS3-ELISA, and the positive rate of BVDV antibody was 24.8%. Compared with commercial kit, the coincidence was 97%. The result suggested that NS3-ELISA was a convenient, rapid method for the diagnosis, antibody level surveillance and epidemiologic investigation of BVDV.