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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 methodology of the clinical diagnosis and epidemiological investigation of spotted fever rickettsia. Methods The prokaryotic expression plasmid of R. Japonica Anhui 120 strain OmpB was constructed, and the recombinant protein was expressed, identified and purified. The indirect ELISA was used as the antigen to detect the specific IgG antibody against T. speciosa in 120 clinical samples. Evaluate the sensitivity and specificity of the method. As a result, OmpB recombinant protein with high expression and high purity was obtained. The established indirect ELISA method has good specificity and sensitivity compared with the diagnostic kit approved by the U.S. Food and Drug Administration. The specificity of the two is 100% , The sensitivity was 96.7%, the coincidence rate was 99.2%, Kappa value was 0.98. Conclusion The OmpB recombinant protein has good immunoreactivity and can specifically detect the IgG antibody of spotted fever rickettsia. The indirect ELISA method can be used as a technical reserve for clinical diagnosis and epidemiological investigation and scientific research.