基于gB蛋白的牛传染性鼻气管炎间接ELISA检测方法的建立

为建立一种针对牛传染性鼻气管炎病毒（Infectious bovine rhinotrachetitis virus,IBRV）的早期快速检测方法，并能判定免疫后是否再次发生感染，本研究利用原核表达IBRV gB蛋白，并通过SDS-PAGE和Western blot检测gB蛋白的表达，并初步建立基于gB蛋白检测IBRV的间接ELISA方法。结果表明：1）对IBRV标准阳性血清，本试验方法的最低检测限度为1∶4 096，说明建立的gB-ELISA检测方法具有较高的敏感性。2）该方法对其他4种常见的牛疫病阳性血清检测均为阴性，不存在交叉反应，表明该方法具有良好的特异性。3）在对200头牛的临床样品的检测中，该方法与IDEXX(IBRV gB X3)抗体检测试剂盒符合率高达97.5%。综上，本研究建立的gB-ELISA检测方法敏感性高，特异性好，该方法为开发一种在感染早期快速对疾病进行诊断，并可以判定疫苗免疫后是否再次发生IBRV感染的试剂盒奠定基础。

Establishment of indirect ELISA method for detection of bovine infectious rhinotracheitis based on gB protein

To establish an early and rapid detection method for bovine infectious rhinotracheitis virus （IBRV） and determine whether infection occurs again after immunization， The IBRV gB protein was expressed in prokaryotic cell and verified by SDS-PAGE and western blot. And an indirect ELISA was preliminarly established. The results indicated that： 1） For IBRV standard positive serum， the minimum detection limit of this experimental method was 1∶4 096， indicating that the established gB-ELISA detection method had high sensitivity. 2） The method was negative to other 4 common bovine blight positive serological tests without cross-reaction， and the method had good specificity. 3） In the detection of 200 clinical samples of cattle， the coincidence rate of this method with IDEXX （IBRV gB X3） antibody detection kit was as high as 97.5%. In summary， the gB-ELISA detection method established in this study had high sensitivity and specificity. This method lays the foundation for developing a rapid diagnostic kit for diseases in the early stages of infection and can determine whether IBRV infection occurs again after vaccine immunization.