日本斑点热立克次体近缘株重组OmpB蛋白抗原性研究

Study on the antigenicity of the recombinant protein of R. Japonica OmpB

目的探讨关于斑点热立克次体的临床诊断及流行病学调查而进行的诊断方法学研究。方法构建R.Japonica Anhui 120 strain OmpB原核表达质粒,表达、鉴定并纯化重组蛋白,用其作为抗原建立间接ELISA检测方法,检测120份临床样本中日本斑点热立克次体特异性IgG抗体,并评价方法的敏感性和特异性。结果获得了高表达、高纯度的OmpB重组蛋白,建立的间接ELISA方法体系与美国食品和药物管理局认可的诊断试剂盒相比有良好的特异性和敏感性,两者的特异性为100%,敏感性为96.7%,符合率为99.2%,Kappa值为0.98。结论 OmpB重组蛋白具有很好的免疫反应性,能特异地检出斑点热立克次体IgG抗体,间接ELISA方法可作为临床诊断的技术储备及流行病调查和科学研究。

Objective To investigate the diagnostic and epidemiological methods of clinical diagnosis and epidemiological investigation of R. Japonica. Methods R. Japonica Anhui 120 strain OmpB prokaryotic expression plasmid was constructed by using gene engineering expression. Identification and purification of recombinant protein was used as antigen to establish indirect ELISA detection method to detect R. Japonica specific IgG antibody. And the sensitivity and specificity of the method will be evaluated in 120 clinical samples. Results The high expression and high purity of OmpB recombinant protein was obtained. Compared with FDA approved diagnostic kit,the established indirect ELISA method system had good specificity and sensitivity. The specificity of both was 100%,the sensitivity was 96. 7%,the coincidence rate was 9. 2%,Kappa value was 0. 98. Conclusion OmpB recombinant protein has a good immune response and can be specifically used to detect R. Japonica IgG antibody. Indirect ELISA method can be used as technical storage for clinical diagnosisor epidemiological investigation and scientific research.