论文部分内容阅读
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ELISA包被抗原检测血清中Cpn IgG抗体和检测36份Cpn-/Ct+IgG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95 kDa的重组蛋白,蛋白质印迹法证明其能与人抗Cpn IgG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1:12 800;检测104例临床血清标本中的IgG抗体,与以色列Savyon Diagnostics公司SeroCP~(TM)IgG ELISA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。
Objective To clone Cpn0585 gene of inclusion body of Chlamydia pneumoniae pneumonia and investigate its value in serodiagnosis. Methods The recombinant plasmid pGEX-6p-2 / Cpn0585 was constructed and induced to express and purify the recombinant protein. BALB / c mice were immunized with indirect ELISA. The antibody titers were detected by ELISA. Antigen Detection Cpn IgG Antibody in Serum and Detection of 36 Cpn- / Ct + IgG Positive Reference Serum. Results A recombinant protein with a relative molecular mass of about 95 kDa was efficiently expressed and purified. Western blotting demonstrated that it reacted with human anti-Cpn IgG. In the immunized BALB / c mice, The detection of IgG antibodies in 104 clinical serum samples was compared with that of SeroCPTM IgG ELISA diagnostic kit from Israel Savyon Diagnostics. The coincidence rate was 98.3%. Conclusions The recombinant protein Cpn0585 has good immunological activity and has high value in the serological diagnosis of Cpn.