沙门氏菌fimY基因的原核表达及禽沙门氏菌抗体检测ELISA研究  

Development and Validation of Procaryotic Expressed Salmonella fimY Protein-based ELISA for Detection of Avian Salmonella Antibody

作　　者：陈小玲[1] 李永清[1] 孙慧玲[1] 徐福洲[1] 章振华[1] 杨兵[1] 

机构地区：[1]北京市农林科学院畜牧兽医研究所,北京100097

出　　处：《中国家禽》2012年第13期18-22,共5页China Poultry

基　　金：北京市科技合同项目(Z090605006009016)

摘　　要：为研发禽沙门氏菌抗体检测方法,采用大肠杆菌系统表达沙门氏菌属保守的菌毛fimY基因,纯化的目的蛋白经Western blot分析,能与免疫鸡血清和感染鸡血清产生特异性反应条带,证明该重组蛋白既有反应原性也有免疫原性。以此纯化蛋白为抗原建立了间接ELISA(fimY-ELISA)。通过检验已知沙门氏菌鸡源血清、相关非沙门氏菌鸡源血清对fimY-ELISA进行评价,发现其在敏感性方面高于快速平板凝集试验(RPA)和自制的脂多糖抗原ELISA(LPS-ELISA)、肠炎沙门氏菌抗原ELISA(SE-ELISA),但在特异性方面不如LPS抗原ELISA;在与RPA的符合率方面与LPS-ELISA、SE-ELISA基本一致。To develop an ELISA for fimY was expressed in E.coli system. The detecting Salmonella antibody in chickens, Salmonella conservative gene antigenicity and immunogenicity of purified fimY protein was proved by Western blot analysis with vaccinated and infected chicken sera. The fimY protein was used to coat ELISA plate and an indirect ELISA was established. To evaluate the specificity and sensitivity of fimY-ELISA, Salmonella and non-Salmonella-positive chicken sera were tested. The results showed that using rapid plate agglutination test （RPA）as gold standard, fimY-ELISA was more sensitive than lipopolysaccharide （LPS） and sonicated Salmonella Enteritidis （SE） antigen-based ELISAs, but which was less specific than LPS-ELISA. In respect to correlation with RPA test, fimY-ELISA,LPS-ELISA and SE-ELISA antigens had no significant difference.

关 键 词：禽沙门氏菌抗体 fimY基因 表达 ELISA 特异性 

分 类 号：S858.315.3[农业科学—临床兽医学]