摘要
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的囊膜蛋白E2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E2-ELISA检测方法,证实该方法的特异性、敏感性和重复性均较好。用所建立的E2-ELISA方法检测从广西各牛场采集的475份血清样品,检出率为24.8%;与商品化试剂合作比较,符合率为95%。结果表明,建立的E2-ELISA方法简便易行,适用大量样本检测,可用于BVDV的抗体水平监测及牛病毒性腹泻的流行病学调查。
An indirect enzyme-linked immunosorbent assay （ELISA） method was developed to detect antibody against of bovine virus diar- rhea virus （BVDV）. The structural protein E2 gene of BVDV was subcloned into prokaryotic vector pET-32a and induced to express. The recombinant protein E2 was used as coating antigen for the establishment of ELISA method, and the reaction conditions were optimized. And the ELISA method showed good results in specificity, sensitivity and reproducibility. Furthermore, four hundred and seventy-five serum sam- pies from several cattle farms of Guangxi were assayed using E2-ELISA, and the positive rate of antibody against BVDV was up to 24. 8%. Compare to commercial kit, the coincidence rate was 95%. The result suggested that the E2-ELISA is a convenient and rapid method for the antibody level surveillance against BVDV and epidemiological investigation of bovine virus diarrhea.
出处
《畜牧与兽医》
北大核心
2014年第10期21-25,共5页
Animal Husbandry & Veterinary Medicine
基金
广西自然科学基金面上项目(2011GXNSFA018096和2012GXNSFAA053074)
广西特聘专家专项(2011B020)
桂科专项(13-3)
