副结核分枝杆菌MAP0862蛋白的表达及在间接ELISA中的初步应用  

Expression of Mycobacterium aviumsubsp.paratuberculosis MAP0862 and Its Primary Application in Indirect ELISA

作　　者：张振[1,2] 冯宇[1] 张阁[2] 朱良全[2] 蒋卉[2] 丁家波[2] 赵鹏[1] 常维山[1] 

机构地区：[1]山东农业大学动物科技学院,泰安271018 [2]中国兽医药品监察所,北京100081

出　　处：《畜牧兽医学报》2016年第4期789-795,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：现代农业人才支撑计划;北京市科技新星(xx2013099)

摘　　要：本研究的目的是探索副结核分枝杆菌特异性蛋白MAP0862在牛副结核病血清学诊断中的作用,建立牛副结核病特异性ELISA诊断方法。将通过原核表达获得的副结核特异性蛋白MAP0862纯化并定量后作为包被抗原,经过一系列条件优化后初步建立了牛副结核病间接ELISA诊断方法。使用牛副结核阳性血清、牛布病阳性血清、牛结核阳性血清、卡介苗免疫牛血清、牛大肠杆菌阳性血清以及健康阴性牛血清对该方法的验证表明其具有良好的特异性。使用该方法对300份临床血清进行检测,结果表明该方法与IDEXX副结核检测试剂盒的符合率为94.3%。基于副结核特异性蛋白质MAP0862建立的间接ELISA诊断方法能有效检测牛副结核病。The objective of the present study was to explore the effect of MAP0862 in sero-diagnosis of Bovine paratuberculosis,and establish a specific ELISA method based on MAP0862.The specific protein obtained by prokaryotic expression was used as coating antigen after purification and quantification,an indirect ELISA was primary established after a series of optimization.The specificity of this method was verified by MAP antiserum,Brucellaantiserum,Bovine tuberculosis（TB）antiserum,BCG antiserum,Escherichia coli antiserum and negative serum.The detection results of 300 serums showed that the coincidence rate of MAP0862 indiret ELISA with commercial kit was 94.3%.The result indicated that the indirect ELISA method primary established based on MAP0862 can detect Bovine paratuberculosis effectively.

关 键 词：牛副结核病 MAP0862蛋白 ELISA 

分 类 号：S855.12[农业科学—临床兽医学]