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.
出处
《中国家禽》
北大核心
2012年第13期18-22,共5页
China Poultry
基金
北京市科技合同项目(Z090605006009016)
