梅毒螺旋体抗原基因的克隆、表达及其在临床检验中的应用

Cloning and expression of Treponema pallidum and application on the clinical detection

目的 克隆、表达并纯化梅毒螺旋体相对分子质量为 47× 10 3 抗原 ,并检测梅毒患者的血清标本。方法 应用基因扩增技术分离此蛋白的全长基因 ,采用基因工程的方法 ,应用融合表达载体pGEX 2T ,重组、克隆得到带有梅毒螺旋体 47× 10 3 特异性抗原基因的重组菌株 ,经大肠杆菌表达系统获得重组融合蛋白。再经亲和层析获得纯品 ,并联合应用ELISA检测梅毒患者的血清标本 30份 ,同时检测非梅毒患者的血清标本 30份。结果 纯化后获得了梅毒螺旋体相对分子质量为 47× 10 3 的特异性抗原。ELISA检测梅毒患者血清结果均阳性。非梅毒患者血清均阴性 ,无非特异性交叉反应。结论 此项方法具有操作简便、准确的特点 。

Objective To study the expression and antigenicity of Treponema pallidum (Tp) outer membrane protein Mr47×10 3. Methods Treponema pallidum outer membrane gene was amplified from the complete genome by PCR, and cloned in the pGEX 2T fused expression vector. Transformed in E.coli , the fused expression vector with Treponema pallidum Mr47×10 3 genome in E.coli was formed. The fusion protein were generated by E.coli fused expression system and purified with affinity chromatography. Correlation with ELISA showed: 30 serum samples of Tp patients were positive, and 30 serum samples of non Tp patients were negative. Results The purified Treponema pallidum Mr47×10 3 antigen could be applied to detect antibody of Tp patients. Conclusion The recombinant of E.coli will be of value for clinical detection of Treponema pallidum.