牛分枝杆菌PPE68与Mb1230以及PPE57和PE-PGRS35的表达纯化及其在牛结核病血清学诊断中的初步应用  

Expression and purification of Mycobacterium bovis PPE68,Mb1230,PPE57 and PE-PGRS35 and their preliminary application in the serodiagnosis of bovine tuberculosis

作　　者：李明[1] 张雅娜 林伟东 隋修锟[1,2] 贾红 侯绍华[1] 姜一曈[1] 房立春[1] 朱鸿飞 鑫婷[1] LI Ming;ZHANG Ya-na;LIN Wei-dong;SUI Xiu-kun;JIA Hong;HOU Shao-hua;JIANG Yi-tong;FANG Li-chun;ZHU Hong-fei;XIN Ting(Institute of A nimal Sciences,Chinese Academy of A gricultural Sciences ,Beijing 100193,China;Molecular Biology, Gembloux Agro-Bio Tech,University of Liege ,Liege 4000,Belgium;Beijing Xinhexiang Technology Co.,Ltd., Beijing 100085,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]比利时列日大学让布鲁农学院分子生物学实验室,列日4000 [3]北京信和翔科技有限责任公司,北京100085

出　　处：《中国兽医科学》2019年第1期20-28,共9页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31602086);“十三五”国家重点研发计划项目(2016YFD0500902,2017YFD0500906);北京市自然科学基金项目(6164039)

摘　　要：为筛选和评估牛分枝杆菌毒力菌株与卡介苗(BCG)基因差异区域(region of differences,RD)蛋白在牛结核病诊断中的效果,本研究表达了牛分枝杆菌RD1区的PPE68、RD9区的Mb1230、R11区的PPE57和RD2区的PE-PGRS35,并纯化得到纯度大于90%的重组蛋白。通过优化条件建立了间接ELISA检测方法,并采用该方法检测了结核病阳性牛和健康牛血清中针对PPE68、Mb1230、PPE57和PE-PGRS35的IgM和IgG抗体水平,评价其在牛结核病血清学诊断中的潜力。结果显示,在结核病阳性牛中,针对PPE68、Mb1230、PPE57和PE-PGRS35的IgG抗体检出率分别为13.3%、43%、50%和30%,IgM抗体检出率分别为30%、10%、36%和33%。结果表明,R11区的PPE57和RD2区的PE-PGRS35有潜力作为牛结核病血清学检测的候选抗原,用于牛结核病的诊断。To screen and evaluate the Mycobacterium bovis specific antigens in regions of differences for the diagnosis of bovine tuberculosis,PPE68(in RD1),PE-PGRS35(in RD2),Mb1230(in RD9)and PPE57(in RD11)were expressed and purified.The purity of the 4 proteins was higher than 90%.An indirect ELISA based on each antigen was established and optimized,and then used to detect the IgM and IgG antibodies against PPE68,PE-PGRS35,Mb1230 or PPE57 in serum from bovine tuberculosis positive and healthy ones,respectively. In result,the detection rates of IgG antibodies against PPE68,Mb1230,PPE57 or pe-pgrs35 in b TB positive cattle were 13.3%,43%,50%and 30%,respectively.While,the detection rates of IgM antibodies were 30%,10%,36%and 33%respectively.The PPE57(in RD11) and PEPGRS35(in RD2) were potential to be used as candidate antigens for the serodiagnosis of bovine tuberculosis.

关 键 词：牛分枝杆菌 血清学检测 重组蛋白 IgG IGM 

分 类 号：S852.618[农业科学—基础兽医学]