布鲁菌omp19原核表达和间接酶联免疫吸附试验检测布鲁菌方法的建立  

Development of a new indirect enzyme-linked immunosorbent based detection assay for Brucella

作　　者：王娜[1,2] 呼和巴特尔[1] 崔步云[2] 

机构地区：[1]内蒙古农业大学,呼和浩特010018 [2]传染病预防控制国家重点实验室感染性疾病诊治协同创新中心中国疾病预防控制中心传染病预防控制所,北京102206

出　　处：《微生物与感染》2014年第1期31-36,共6页Journal of Microbes and Infections

基　　金：国家高科技研究发展计划(863计划)(2007AA02Z410);国家自然科学基金(81160355)

摘　　要：本研究原核表达并纯化了布鲁菌脂蛋白Omp19,并通过蛋白免疫印迹法对其免疫原性进行验证。采用Omp19作为包被抗原,建立检测羊种布鲁菌的间接酶联免疫吸附试验(iELISA),检测90份羊血清样本,并与标准血清凝集试验(SAT)比较,用SPSS软件进行数据分析。结果显示,iELISA与SAT的阳性符合率为95.12%,阴性符合率为67.35%,Kappa值为0.607 7。对2种方法进行McNemar卡方检验,结果有显著性差异(P<0.05)。本研究建立的iELISA与SAT的总符合率达80%,可作为羊种布鲁菌病血清学诊断的备用方法。Brucella protein Omp19 was expressed and purified .Its immunogenicity was further validated by Western blotting .An indirect enzyme-linked immunosorbent assay （iELISA） for the detection of Brucella melitensis （B . melitensis） was established using the purified recombinant protein Omp 19 as coating antigen . Serum samples （n=90） from sheep were simultaneously tested by iELISA and standard serum agglutination test （SAT） ,and the data was analyzed by SPSS software .Compared with SAT ,the positive coincidence rate of the Omp 19 based iELISA was 95 .12% and the negative coincidence rate was 67 .35% .Kappa value was 0 .607 7 .McNemar chi-square test showed that there was significant difference between the two methods （ P〈0 .05） . The total diagnosis coincidence rate was 80% . The established iELISA could be used as an alternative method for serological diagnosis of brucellosis .

关 键 词：布鲁菌 omp19 间接酶联免疫吸附试验 标准血清凝集试验 

分 类 号：R446.6[医药卫生—诊断学]