牛传染性鼻气管炎病毒DQ株gD基因表达及其间接ELISA方法的建立和初步应用  

Establishment and preliminary application of an indirect ELISA assay to detect antibodies against IBRV-DQ using expressed gD proteins

作　　者：董华兴[1] 侯喜林[1] 谢金鑫[1] 张佩鑫[1] 柳强[1] 刘鹏[1] 王延涛[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《中国兽医杂志》2011年第2期3-5,共3页Chinese Journal of Veterinary Medicine

基　　金：大庆市科技局资助项目(SGG2006-010)

摘　　要：以原核表达的牛传染性鼻气管炎病毒(IBRV)重组gD蛋白作为包被抗原,建立检测IBRV抗体的间接ELISA方法。构建pET30a-gD重组质粒,在大肠杆菌中表达IBRV重组gD蛋白,Western-blot检测该重组蛋白具有良好的免疫活性。通过方阵试验确定了抗原的最适包被浓度为2.5μg/mL,血清最佳稀释倍数为1∶200,酶标二抗最佳稀释倍数为1∶5 000,与病毒中和试验、全病毒ELISA方法的检测结果符合率分别为87.5%、92.5%。用该法对黑龙江部分地区采集到的863份牛血清样品进行检测,结果阳性率为82.27%。本试验建立的间接ELISA方法可用于IBR的检测和流行病学调查。An indirect ELISA was developed to detect antibodies against IBRV using recombinant gD protein of infectious bovine rhinotracheitis virus（IBRV）.The prokaryotic expression system of pET30a-gD expressing the IBRV recombinant gD proteins was used.Western blot was employed to detect the recombinant proteins.Through the matrix experiments,the optimal coating antigen concentration was 2.5μg/mL,and the optimal serum dilution was 1∶200,and the optimal dilution of HRP-labeled rabbit anti-bovine IgG was 1∶5 000.In comparison to the VN Test and the whole virus ELISA,the agreement rate of gD-ELISA is 87.5% and 92.5%,respectively.The 863serum samples from Heilongjiang province were detected by this assay.The positive rate was 82.27 %.This ELISA can be used for epidemiological survey for infectious bovine rhinotracheitis.

关 键 词：牛传染性鼻气管炎病毒 重组gD蛋白 间接ELISA 抗体检测 

分 类 号：S852.659.6[农业科学—基础兽医学]