双抗体夹心ELISA检测羊肠道病毒抗原方法的建立及病原流行病学调查

发布时间：2018-06-21 08:40

  本文选题：羊肠道病毒 + CEV-JL　； 参考：《中国兽医学报》2017年04期

【摘要】：应用本实验室分离的国内首株羊肠道病毒(CEV)-JL14VP1重组蛋白制备的兔源多克隆抗体和抗CEV-JL14病毒的单克隆抗体,建立了检测CEV抗原的双抗体夹心ELISA方法,并对吉林省和内蒙古地区羊群感染CEV进行了病原流行病学调查。方阵试验确定了CEV VP1鼠源IgG捕获抗体的最佳包被量为0.2μg/孔,酶标抗体的最佳稀释倍数为1∶1 000。对大量CEV阴性粪便样品进行检测与统计学处理,确定了双抗体夹心ELISA检测CEV的判定标准为样品D490值≥0.216判定为阳性。特异性、敏感性和重复性试验结果表明,所建立的检测CEV抗原方法具有特异、敏感、快速和重复性好等特点。以建立的双抗体夹心ELISA方法,对吉林省和内蒙古不同地区的羊群粪便样品进行了检测,发现不同地区的羊群都存在着严重的CEV感染,感染率高达12%~100%。本研究为今后CEV感染的诊断、检疫及防制提供了有效的手段和流行病学理论依据。
[Abstract]:Using rabbit polyclonal antibody and monoclonal antibody against CEV-JL14 virus, the first sheep enterovirus recombinant protein in China, a sandwich Elisa method for detecting CEV antigen was established by using rabbit polyclonal antibody and monoclonal antibody against CEV-JL14 virus. The pathogen epidemiology of sheep infection with CEV in Jilin province and Inner Mongolia was investigated. The best encapsulation of IgG capture antibody of CEV VP1 mouse source was 0.2 渭 g / well and the best dilution multiple of enzyme labeled antibody was 1:1 000 by square array test. A large number of CEV-negative fecal samples were detected and statistically analyzed. The standard of double antibody sandwich Elisa for CEV detection was determined to be that the sample D490 鈮，

本文编号：2047925