牛传染性鼻气管炎抗体的纳米胶体金斑点免疫渗滤法检测

Studies on nano-colloidal gold dot immuno-filtration assay for detection of antibodies of infectious bovine rhinotracheitis

将无血清细胞培养的牛传染性鼻气管炎病毒(IBRV)抗原包被于硝酸纤维素膜,加入待检血清样品后,利用纳米胶体金标记的山羊抗牛IgG显色,建立了检测IBR抗体的纳米胶体金斑点免疫渗滤法检测试纸盒。整个试验过程仅需5min即可判断结果,与蓝舌病、牛赤羽病、牛地方流行性白血病、牛病毒性腹泻黏膜病、猪瘟、猪伪狂犬病和猪细小病毒病的阳性血清不发生交叉反应。将该法与用于IBR抗体检测的ELISA方法同时对300份临床牛血清样品进行IBR抗体检测,结果二者的阳性符合率达94.4%。结果表明,该法具有特异、敏感、快速可靠、效果直观、结果容易判断的特点,非常适用于IBR的早期诊断和流行病学调查。

The infectious bovine rhinotracheitis virus（IBRV） antigen from the virus culture without serum was coated on nitrocellulose（NC）membrane,then the test serum was added to NC membrane,and the color reaction was done with nano-colloidal gold-conjugated goat-anti-bovine IgG.The nano-colloidal gold dot immuno-filtration assay（DIFA） of detecting antibody of IBR was developed by this procedure.The whole procedure could be finished in 5 minutes.It was specific and had no cross-reaction with positive sera infected with bluetongue virus,bovine akabane virus,bovine leukaemia virus,bovine viral diarrhea-mucosal virus,hog cholera virus,swine pseudorabies virus or porcine parvovirus.The coincidence rate of the results of simultaneous determination of 300 sera with ELISA and DIFA was 94.4% for antibody of IBR.The results showed that DIFA is a specific,sensitive,visual,rapid and simple detecting method,which can be applied for earlier diagnosis and epidemiological investigation of IBR.