梅毒螺旋体TpN17抗原全长表达纯化和ELISA检测  

Cloning and Expression of the Full Length TpN17 Antigen of Treponema Pallidum and the Detection in ELISA

作　　者：刘灵辉 

机构地区：[1]广州万孚生物技术有限公司,广东广州510641

出　　处：《中国皮肤性病学杂志》2009年第1期9-10,19,共3页The Chinese Journal of Dermatovenereology

基　　金：广东省产学研项目(编号2006A11601010);2007年广州市科技计划项目

摘　　要：目的在大肠埃希菌中重组表达梅毒密螺旋体TpN17抗原,用其建立梅毒诊断血清学方法。方法酶切与PCR联用,合成全长TpN17基因,亚克隆后克隆到pET-HIS质粒并转化到大肠杆菌BL21株中表达。亲和层析纯化。用纯化的rTpN17包被微孔板,ELISA方法检测临床8份梅毒阳性血清和4份梅毒阴性血清。结果获得了重组表达并纯化的TpN17,ELISA检测结果表明重组蛋白能被梅毒患者阳性血清所识别,特异性100%,灵敏度100%。结论该重组蛋白具有良好的免疫反应性,能鉴别梅毒阳性血清,有望进一步应用于梅毒感染的血清学检测。Objective To express the recombinant TpN17 protein of Treponema pallidum （TP）in E. Coli, and to develop ELISA for serological diagnosis of syphilis. Methods Synthesis of full length Gene TpN17 by enzyme companied with PCR, subclone and then cloned into the expression vector pET - HIS, and expressed in E. coli BL21. Purified with Ni - NTA affinity chromatography columns. Micro well plate was coated with purified rTpN17, and then 4 negative and 8 positive sera was tested by ELISA. Results Gained purified rTpN17 protein, and the result of ELISA with the sensitivity of 100% and specificity of 100%. Conclusion The rTpN17 protein can be used for development of ELISA for serological diagnosis of syphilis.

关 键 词：梅毒密螺旋体 重组蛋白 酶联免疫吸附法 

分 类 号：R759.1[医药卫生—皮肤病学与性病学]