期刊论文

中文

中华检验医学杂志

1009-9158

2005

28

10

996-998

10.3760/j:issn:1009-9158.2005.10.008

湖南省教育厅重点研究项目

本校为第一机构

目的克隆、表达梅毒螺旋体(Tp)外膜蛋白Tp0453,建立间接ELISA法,探讨其在梅毒血清学诊断中的应用.方法构建重组质粒pQE32/Tp0453,IPTG诱导表达,SDS-PAGE和免疫印迹鉴定表达结果;Ni-NTA亲和层析法纯化重组蛋白,间接酶联免疫吸附试验(ELISA)检测人血清中特异性IgG抗体.结果成功构建了重组质粒pQE32/Tp0453;SDS-PAGE检测诱导产物显示有一分子量约为32000的特异蛋白带,免疫印迹检测其只与Tp IgG抗体具有阳性反应;重组蛋白用作ELISA包被抗原检测Tp阴阳性参比血清,特异度和灵敏度均为100%;检测患者血清中特异性IgG抗体结果与TPPA法符合率为98.2%.结论表达的Tp0453重组蛋白具有较好的免疫反应活性,可望...

Objective To clone, and express Tp0453 outer membrane protein of Treponema pallidum and develop an indirect ELISA for sero diagnosis of syphilis. Methods The immuno-dominant epitope of Tp0453 was amplified by PCR and subcloned into the expression vector pQE32. The recombinant protein was expressed in E. coli M15 and purified with Ni-NTA affinity chromatography columns. Indirect ELISA was developed to detect the antibody to Tp in human sera. Results 60 control sera was tested by ELISA. The sensitivities was 100% （30/30）, and the specificities ...