Cpn0585蛋白的免疫活性及血清学诊断初步评价

目的克隆肺炎嗜衣原体包涵体膜蛋白Cpn0585基因，初步探讨其在血清学诊断中的应用价值。方法构建pGEX-6p-2／Cpn0585重组质粒，诱导表达并纯化重组蛋白，免疫BALB／c小鼠，间接酶联免疫附试验（ELISA）检测小鼠血清抗体滴度，以重组蛋白作为ELBA包被抗原检测血清中CpnIgG抗体和检测36份Cpn-／Ct＋IsG阳性参考血清。结果高效表达和纯化出一相对分子质量约为95kDa的重组蛋白，蛋白质印迹法证明其能与人抗CpnIgG抗体发生反应；在被免疫的BALB／c小鼠体内，特异性IgG抗体的滴度为1：12800；检测104例临床血清标本中的IgG抗体，与以色列SavyonDiagnostics公司SeroCPTMIgGELISA诊断试剂盒的检测结果进行比较，符合率为98．3％。结论表达的Cpn0585重组蛋白具有良好的免疫活性，在Cpn的血清学诊断中具有较高的应用价值。

Preliminary Evaluation of Immunocompetence and Serological Diagnosis of Cpn0585 Protein

Objective To clone and express the Cpn0585 gene from Chlamydophila pneumonia e. and then evaluate the clinical value of the recombinant protein in serodiagnosis. Methods The pGEX-6p-2/Cpn0585 recombinant plasmid was contructed, and then expression of recombinant protein was induced. The expressed recombinant protein was identified by SDS-PAGE. BALB/c rats were immunized with the purified protein;indirect ELISA was applied to measuring its immunogenieity; and its immunoreactivity was analyzed by reacting with Cpn-specific IgG antibody in sera. Cpn IgG and Chlamydozoa trachomatis positive sera were detected with indirect ELISA. Results The recombinant protein with relative molecular weight about 95 kDa was expressed and purified effectively. Western blot analysis proved that the recombinant protein could react with human anti Cpn IgG specifically. The titer of the specific IgG antibodies in the immuned BALB/c rats was above 1：12 800. The compliance rate between the indirect ELISA test and SeroCPTM lgG ELISA kits to104 patients sera were 98. 3%. Conclusion The expressed recombinant protein of Cpn0585 from Cpn shows excellent irnmunoc-ompetence. It is suitable for preparation of serodiagnostic EIISA reagent for Cpn infection.