微小隐孢子虫病毒衣壳蛋白抗体检测ELISA方法的建立  

Development of indirect ELISA for detection of the antibody against Cryptosporidium parvum virus

作　　者：刁玉梅[1] 宫鹏涛[1] 李巍[1] 苏利波[1] 黄祥盛[1] 高华义[1] 李赫[1] 胡进平[1] 李建华[1] 张西臣[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国预防兽医学报》2011年第7期537-540,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技支撑项目(2007BAD40B05);吉林省科技发展计划项目(20100160)

摘　　要：为建立快捷的含病毒隐孢子虫的检测方法,本研究利用已构建的微小隐孢子虫病毒衣壳蛋白重组表达质粒表达并纯化重组蛋白,建立以该重组衣壳蛋白为包被抗原检测抗体的间接ELISA检测方法。经优化后间接ELISA的最佳条件为:抗原包被浓度为0.25μg/孔,被检血清稀释度为1∶200,酶标二抗稀释度为1∶2000,封闭液为含5%脱脂奶粉的PBST溶液。该方法检测安氏隐孢子虫阳性血清,柔嫩艾美尔球虫阳性血清,蓝氏贾第虫阳性血清,弓形虫阳性血清与微小隐孢子虫阳性结果均为阴性。该方法灵敏度为1∶1600,特异性较强且具可重复性,可用于含病毒隐孢子虫的抗体检测及流行病学调查。To develop an indirect ELISA for detection of the antibody against Cryptosporidium parvum virus（CSpV）, recombinant protein of CSpV capsid protein was purified as coating antigen. The optimal antigen concentration for coating was 0.25 μg each well, and the optimal dilutions of the sera of C. parvum and enzyme-labeled the second antibody were 1∶200 and 1∶2 000 respectively, blocking agent was the 5% skim milk PBST by chessboard titration. The developed indirect ELISA had no cross reaction with positive mouse serum against C. parvum, C. andersoni, E. tenella and T. gondii. It showed high sensitivity of 1∶1 600. The indirect ELISA for detection of the antibody against C. parvum virus might be used for the diagnosis and epidemiological investigation of the virus infected Cryptosporidium.

关 键 词：微小隐孢子虫 dsRNA病毒 衣壳蛋白 间接ELISA 

分 类 号：S852.72[农业科学—基础兽医学]