Tp0259-PCR 的建立及其在一期梅毒诊断中的初步应用∗

摘要

Objective To explore the significance of established Tp0259-PCR in diagnosing primary syphilis.To provide basis for establishing new PCR diagnosis method of Treponema pallidum.Methods Specific primers were designed by the gene sequences of Tp0259 which was obtained from Genbank.Tp0259-PCR was established under the optimization reaction condition.Using the new construction of amplification method to detect the common pathogenic microorganism in urinary and reproductive tract infec-tion.42 clinical secretion specimens suspected primary syphilis were tested by Tp0259-PCR.Serum specimens from the same pa-tients were tested by TPPA.Results Only specific amplicons could be found in amplifying the Treponema pallidum by Tp0259-PCR.The detection rate was 73.8% using Tp0259-PCR which was apparently higher than 57.1% using TPPA(P <0.05 ).The specificity of the established method was 100% comparing with TPPA.Conclusion The established method of Tp0259-PCR has high sensibility and specificity.This method might be better than serological method in diagnosis of primary syphilis.but its clinical diagnosis value need to be repeatedly verified by more clinical specimens.%目的：建立聚合酶链反应(PCR)方法扩增梅毒螺旋体(Tp)0259基因,探讨其在一期梅毒诊断中的意义,为建立新的梅毒 PCR 诊断方法提供依据。方法从 Genbank 获取 Tp0259基因序列设计引物,优化反应条件,建立 Tp0259-PCR 方法；同时通过新构建扩增方法检测泌尿生殖道感染常见病原微生物；以建立的 Tp0259-PCR 方法检测临床42例确诊一期梅毒患者的分泌物,同时用血清学方法 Tp 明胶颗粒凝集试验(TPPA)平行检测待测者血清中抗 Tp 抗体。结果Tp0259-PCR 只在扩增 Tp 时有特异性产物；其他病原微生物未见扩增条带；检测确诊一期梅毒患者标本,Tp0259-PCR 检出率(73.8%)明显高于 TPPA(57.1%)(P <0.05),与 TPPA 相比,Tp0259-PCR 特异度为100%。结论建立的 Tp0259-PCR 法具有高敏感性和高特异性,初步应用显示其对一期梅毒诊断优于血清学方法,但其临床诊断价值有待对大量临床标本进行反复验证。