抗猪肺炎支原体IgA间接ELISA检测方法的建立  

Establishment and Application of Indirect ELISA for Mycoplasma hyopneumoniae IgA Detection

作　　者：逯晓敏[1,2] 冯志新[1] 刘茂军[1] 吴叙苏[1] 甘源[1] 张映[2] 邵国青[1] 

机构地区：[1]江苏省农业科学院兽医研究所农业部动物疫病诊断与免疫重点开放实验室国家兽用生物制品工程技术研究中心,南京210014 [2]山西农业大学动物科技学院,太谷030801

出　　处：《畜牧兽医学报》2009年第10期1569-1574,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：中国博士后基金(20070421022);江苏省农业三项工程(SX2007082);江苏省自然科学基金(BK2007711)

摘　　要：为了更准确地对猪肺炎支原体感染及免疫状况进行检测,作者拟建立抗猪肺炎支原体IgA间接ELISA检测方法。在比较了猪肺炎支原体全菌抗原和粘附因子P97R1原核表达蛋白后,选择后者作为包被抗原;以羊抗猪IgA为二抗,兔抗猪IgG-HRP作为酶标三抗,并分别对包被抗原的浓度、样品的孵育时间、二抗与三抗的最佳稀释度和作用时间以及显色液的作用时间作了优化。最终建立了稳定而特异的抗猪肺炎支原体IgA的间接ELISA检测方法。批间与批内试验结果证明该方法具有很好的稳定性。初步应用表明,该方法可用于猪肺炎支原体感染或免疫状态的临床监测。To detect the infection or immune state of Mycoplasma hyopneumoniae （Mhp） exactly, an indirect ELISA assay for anti-Mhp IgA detection was developed. In this study, P97 R1 protein of Mhp was chosen as coating antigen, goat anti pig IgA and rabbit anti pig IgG-HRP were used as second and third antibody, respectively. The concentrations of coating antigen, second and third antibodies were optimized as well as their working time. As a result, a stable and specific indirect ELISA assay for anti-Mhp IgA detection was developed. And the developed method could be used for Mhp infection and immune state monitoring.

关 键 词：猪肺炎支原体 IGA 间接ELISA 

分 类 号：S852.62[农业科学—基础兽医学]