牛病毒性腹泻病毒重组E2蛋白PPA-ELISA方法的建立及临床应用  

Development and Application of PPA-ELISA for Detecting Bovine Viral Diarrhea Virus Antibody Using E2 Recombinant Protein

作　　者：柴顺秀 马花 韩英 赵隆寿 

机构地区：[1]青海省湟中县动物卫生监督所,青海湟中811600

出　　处：《动物医学进展》2013年第6期106-110,共5页Progress In Veterinary Medicine

摘　　要：用牛病毒性腹泻病毒(BVDV)重组E2蛋白为包被抗原,辣根过氧化物酶标记葡萄球菌A蛋白(PPA)为二抗,建立了检测BVDV血清抗体的E2-PPA-ELISA方法。研究选取特异性良好的E2抗原,确定了最佳工作条件为:E2蛋白抗原的最适包被质量浓度为16.0μg/mL,酶标二抗的工作浓度为1∶2 000,血清稀释度为1∶40时效果最佳。通过重复性试验、特异性试验和敏感性试验证明,与用BVDV全病毒为抗原的IDEXX ELISA试剂盒相比,建立的E2-PPA-ELISA抗体检测方法具有较好的重复性、敏感性和特异性。对285份不同地区采集的血清样本进行检测,阳性检出率为33.4%,与IDEXX ELISA试剂盒的检出率无明显差异。表明该方法为BVDV的快速诊断和流行病学调查提供一种快速、简便的血清学诊断方法,适合规模化养牛场BVDV的抗体监测以及流行病学调查。To develop a method to detect antibodies against BVDV,an PPA-ELISA was established by the recombinant E2 protein antigen expressed in E.coli prokaryotic expression system.The optimal coating concentration of E2 recombinant protein was 16.0 μg/mL.The dilution of serum sample was 1∶40 and optimal concentration of HRP-SPA labeled antibody IgG was 1∶2 000.Compared with the routine IDEXX ELISA test kit with the whole virus,the established method has good specificity,sensitivity and repeatability.In addition,285 serum samples were detected by the E2-PPA-ELISA and the IDEXX BVDV Antibody Test Kit.The results showed that the BVDV antibody positive rate was 33.4%.Therefore,the E2-PPA-ELISA was practical for serological diagnosis of BVDV and antibody surveillance.

关 键 词：牛病毒性腹泻病毒 重组E2蛋白 PPA-酶联免疫吸附试验 

分 类 号：S852.653[农业科学—基础兽医学]