免疫PCR法诊断梅毒螺旋体抗原和抗体的方法学研究

目的建立免疫PCR方法检测梅毒螺旋体抗原和梅毒螺旋体抗体两种方法学，并比较两种方法学的敏感性、特异性和重复性。方法选择2011年我院梅毒血清学ELISA法检测者1520例并梅毒分期；建立免疫PCR法检测梅毒螺旋体抗原和梅毒螺旋体抗体两种方法，然后对梅毒螺旋体血清学阳性标本10倍系列稀释，比较免疫PCR法敏感性；同时采用正常健康血清标本观察免疫PCR法特异性；最后采集梅毒阳性血液分装成两份标本，监测两种免疫PCR法试验批内和日间重复性。结果1520例患者中，梅毒患者136例。梅毒患者中女性占77．2％（105／136）．孕妇占19．1％（26／136）。梅毒分期以一期梅毒感染为主，与其他期比较具有统计学意义（P〈0．05）。IPCR-Ag法检测梅毒螺旋体抗原敏感性是ELISA法的10。倍，TPPA和TRUST法的10^6。IPCR-Ab法检测梅毒螺旋体抗体敏感性是ELISA法的10。倍，TPPA和TRUST法的10^5。两种免疫PCR法测检查正常健康人群梅毒螺旋体结果均为阴性。IPCR-Ab法试验批内CV为11．6l％，日间CV为18．12％；IPCR-Ag法试验批内CV为18．61％，日间CV为22．24％，两者相比具有统计学意义（P〈0．05）。结论免疫PCR法检测梅毒螺旋体具有高度的敏感性和特异性，但重复性较差。

Study on treponema pallidum specific antigen and antiboby by immuno-PCR

Objective To establish and study the technology to detect Treponema pallidum specific antigen and antiboby by immuno-PCR,and compare with sensibility,specificity and reproducibility.Methods 1 520 cases with syphilis were provided by ELISA and by stages.The technology of detecting Treponema pallidum specific antigen and antiboby by immuno-PCR was established,and sensibility by 10 double time liquor natrii chloridi isotonicus was compared,and specificity by normal specimen was observed,the reproducibility was studied by batch and days.Results The rate was 8.9% on 136 positive syphilis cases detected in 1 520,and 77.2% in femme,and 19.1% in pregnant woman.The primary syphilis possess major cases.The sensitivity of immuno-PCR diagnostic antigen method was 104 times higher than that of routine ELISA method,and 106 times higher than TPPA and TRUST.The sensitivity of immuno-PCR diagnostic antiboby method was 103 times higher than that of routine ELISA method,and 105 times higher than TPPA and TRUST.The specificity was all by immuno-PCR in normal specimen.The bath coefficient of differentiation was 11.61% by immuno-PCR diagnostic antigen,and 18.61% by immuno-PCR diagnostic antiboby.The days coefficient of differentiation was 18.12% by immuno-PCR diagnostic antigen,and 22.24% by immuno-PCR diagnostic antiboby.Conclusion It indicates that immuno-PCR method possesses fairly high sensitivity and specificity and has much discrepancy of reproducibility.