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Position: Home > Articles > Establishment of an indirect ELISA diagnostic method based on the outer membrane protein A(OmpA) of bovine Chlamydophilia abortus Chinese Veterinary Science 2010 (7) 696-700

奶牛流产衣原体OmpA抗原间接ELISA检测方法的建立

作  者:
宋竹青;邱昌庆;周继章;郑福英;蔺国珍
单  位:
山东农业大学动物科技学院;甘肃农业大学动物医学院;中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室;中国农业科学院兰州兽医研究所
关键词:
牛;流产衣原体;重组OmpA;间接酶联免疫吸附试验
摘  要:
以奶牛流产衣原体外膜蛋白A(OmpA)作为抗原,筛选最佳反应条件,建立了奶牛衣原体病间接ELISA检测方法。结果显示,OmpA的最佳包被浓度为1∶320;血清最佳稀释度为1∶40;酶标二抗的最佳工作浓度为1∶2000;抗原的最适包被条件为37℃1h加4℃过夜;ELISA的阴性、阳性临界值为0.391。用本方法和衣原体间接血凝试验检测奶牛血清田间样品206份,两者的符合率为95.3%。本试验建立的OmpA-ELISA方法具有良好的特异性、敏感性和稳定性,可为牛衣原体病的诊断及流行病学调查提供有效工具。
译  名:
Establishment of an indirect ELISA diagnostic method based on the outer membrane protein A(OmpA) of bovine Chlamydophilia abortus
作  者:
SONG Zhu-qing 1,2,QIU Chang-qing 1,ZHOU Ji-zhang 1,ZHENG Fu-ying 1,LIN Guo-zhen 1,CAO Xiao-an 1,WANG Guang-hua 1,GONG Xiao-wei 1,WEI Yan-ming 2(1.Key Laboratory of Veterinary Public Health of the Ministry of Agriculture/State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;2.Gansu Agricultural University,Lanzhou 730070,China)
关键词:
bovine ;Chlamydophilia abortus;recombinant OmpA;indirect ELISA
摘  要:
Bovine Chlamydophilia abortus OmpA was served as antigen for ELISA,after the optimal reaction conditions were selected,and an indirect ELISA was developed to detect bovine chlamydiosis.The optimal concentration of OmpA,serum and anti-bovine IgG-HRP was 1∶320,1∶40,1∶2000 respectively,the best coating condition was 37℃ 1h and 4℃ overnight,and the critical value of the indirect ELISA was 0.391.Comparison between OmpA-ELISA and the indirect hemagglutination test showed the two methods had 95.3% coincidence by detecting 206 field dairy cattle serum samples.The indirect OmpA-ELISA was a specific,sensitive,reliable and effective tool for the diagnosis and epidemiologic investigations of bovine chlamydiosis.

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