结核分枝杆菌蛋白Rv0315的克隆表达及其抗原性研究

目的评价结核分枝杆菌重组蛋白Rv0315在结核病血清学诊断中的价值。方法采用常规分子克隆方法获得重组Rv0315蛋白,酶联免疫吸附试验（ELISA）检测84份确诊结核病人血清和48份健康人血清中相应的抗结核抗体水平,并与结核分泌蛋白ESAT-6的检测结果进行比较。结果成功构建了重组蛋白Rv0315,其在E.coli BL21plysS（DE3）中主要以可溶性形式表达,分子量（30.3kDa）与预期相符,ELISA血清学活性评估显示其特异性和敏感性分别为94.0%（45/48）和35.7%（30/84）。结论重组蛋白Rv0315有望成为新的结核病血清学诊断候选抗原。

Cloning and Expression of M. tuberculosis Rv0315 protein and evaluation of its immunogenicity

Objective To evaluate the value of recombinant Rv0315 protein in serodiagnosis of TB. Methods The gene encoding Rv0315 protein was amplified and cloned and confirmed by sequencing. The expression vector pET32a-Rv0315 was constructed and expressed in E. coli BL21 plysS （DE3） strain. Recombinant Rv0315 protein was purified by Ni-NTA puri- fication system and was used as antigen to screen the sera of tuberculosis （TB） patients （n=84） and healthy controls （n=48） by ELISA, compared with ESAT-6 protein to evaluate the value in serodiagnosis of TB. Results The ORF Rv0315 was ex- pressed mainly as soluble protein in E. coli BL21 plysS （DE3）, with the relative molecular weight of 30.3kDa in line with expec- tations. The immunogenie evaluation of recombinant Rv0315 protein by ELISA suggested that its specificity and sensitivity re- spectively reached 94.0 %（45/48） and 35.7 %（30/84）. Conclusion The recombinant Rv0315 protein is expected to be a new candidate antigen for the serodiagnostic research in TB.