免疫PCR技术检测梅毒螺旋体特异性蛋白的意义

Significance of PCR in detection of immune-specific protein of Treponema pallidum

目的探讨免疫PCR技术检测梅毒螺旋体特异性蛋白TpN47的意义。方法应用PCR法、甲苯胺红不加热血清试验(TRUST)法和酶素螺旋体明胶颗粒凝集试验(TPPA)法进行检测,比较三者的敏感性和特异性。分别用棉拭子取患者泌尿道处渗出液或血清为标本,所有操作严格按照试剂使用说明书进行,PCR方法用细菌基因组DNA提取试剂盒提取TP基因组DNA,设计TpN47基因的引物,取PCR扩增产物进行电泳检测,然后进行T-A克隆、亚克隆及测序,进行提纯、分析,TRUST法和TPPA法按常规进行。结果单用三种检测方法 ,检出率无显著差异(P>0.05),三种方法的灵敏度、特异性不同,PCR法优于TRUST法和TPPA法(P<0.05)。结论免疫PCR技术检测TpN47抗原具有较高的敏感性和特异性,值得临床重视,对有效控制梅毒的蔓延和胎传梅毒的发生具有重要意义。

Objective To study the significance of immunological detection of PCR for Treponema-specific protein TpN47.Methods The sensitivity and specificity of PCR method,TRUST method and TPPA method were studied and compared.Cotton swabs were used to obtain exudates from urinary tract of patients or serum as samples for detection,all procedures were carried out in strict accordance with instructions of reagents,PCR method was applied with bacterial genomic DNA extraction kit for extraction of genomic DNA of TP,primer was designed for TpN47 gene,PCR expansion yielding material was tested by electrophoresis,then TA cloning,subcloning and sequencing,purification and analysis were carried out.TRUST method and TPPA method were performed in accordance with the routine procedures.Results Three detection methods were separately applied,and there was no difference in detection rates among these three methods.The sensitivity and specificity of these three methods were different,PCR method was better than TRUST and TPPA methods.Conclusion The detection of TpN47 antigen by PCR has higher sensitivity and specificity,it is worthy to be emphasized,and it is very important in effective control of the dissemination of syphilis and its fetal transmission.