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目的探讨肺炎嗜衣原体(CPN)蛋白酶样活性因子(CPAF)重组蛋白在CPN感染实验室诊断中的应用价值。方法构建CPAF免疫优势区基因重组质粒,诱导表达并纯化重组蛋白,分析其抗原特异性;间接ELISA法检测CPN参考血清、临床血清标本中的特异性IgM和IgG抗体,以及沙眼衣原体(Ct)阳性血清。结果高效表达和有效纯化出一相对分子质量(Mr)约为5.13×103的特异性重组蛋白;间接ELISA法测定用其免疫兔血清中的特异性IgG和IgM抗体效价,分别高达1∶16 000和1∶8 000;检测Cpn IgM和IgG参考血清,阴性和阳性结果符合率均为100.0%;与“金标准”方法MIF比较,检测300份呼吸道感染患者抗血清,IgMI、gG符合率分别为98.3%、99.0%;检测Ct阳性参考血清和阳性临床血清标本,无阳性结果。结论表达的CPN CPAF免疫优势区重组蛋白具有良好的抗原性,在CPN感染的实验室诊断中具有较高的应用价值。
Objective To investigate the value of CPN protease-like factor (CPAF) recombinant protein in the laboratory diagnosis of CPN infection. Methods The recombinant plasmid of CPAF immunodominant region was constructed and the recombinant protein was induced to express and purify the recombinant protein. The indirect ELISA was used to detect the specific IgM and IgG antibodies in CPN reference serum and clinical serum samples, and the positive of C. trachomatis (Ct) serum. The results showed that a specific recombinant protein with molecular weight (Mr) of about 5.13 × 103 was efficiently expressed and efficiently purified. The titer of specific IgG and IgM antibodies in rabbit serum immunized by indirect ELISA was 1:16 000 and 1: 8000, respectively. The coincidence rates of negative and positive results of both Cpn IgM and IgG reference sera were 100.0%. Compared with the “gold standard” method MIF, the sera of 300 respiratory infections were detected by antiserum, IgMI and gG The rates were 98.3% and 99.0%, respectively. There was no positive result in detecting Ct positive reference serum and positive clinical serum samples. Conclusions The expressed CPN CPAF immunodominant region recombinant protein has good antigenicity and has high value in the laboratory diagnosis of CPN infection.