半巢式聚合酶链反应扩增全血中梅毒螺旋体polA基因  

Amplification of the DNA Polymerase I Gene of Treponema Pallidum from Whole Blood of Persons With Syphilis Using a Semi-nested PCR Assay

作　　者：曾铁兵[1] 吴移谋[1] 赵飞骏[1] 刘双全[2] 余敏君[1] 

机构地区：[1]南华大学医学院病原生物学研究所,湖南衡阳421001 [2]南华大学第一附属医院

出　　处：《南华大学学报（医学版）》2009年第6期648-650,702,共4页Journal of Nanhua University(Medical Edition)

基　　金：南华大学博士科研启动基金(2007XQD29)

摘　　要：目的探讨半巢式聚合酶链反应(PCR)扩增梅毒螺旋体(Tp)的DNA多聚酶Ⅰ基因(polA)以探讨检测全血标本中微量Tp的可行性。方法应用半巢式PCR和常规PCR分别扩增165例可疑梅毒及非梅毒患者全血中Tp的polA,与血清学方法比较,探讨半巢式PCR方法在扩增全血中Tp DNA的意义。结果待测的165例标本中,与血清学方法比较,半巢式PCR检测Tp的敏感性和特异性分别为62.7%和95.9%,两者符合率为82.4%;两种PCR方法检测结果差异有显著性(P<0.01),半巢式PCR敏感性远高于常规PCR。结论半巢式polA PCR检测全血中Tp较常规PCR灵敏,是血清学方法诊断梅毒的有效补充,但其检测的灵敏度仍有待进一步提高。Objective To investigate the contribution of a semi - nested polA PCR assay for the detection of extremely low numbers of treponema pallidum （Tp） in whole blood of persons with syphilis. Methods Routine PCR and semi - nested PCR assays were performed to amplify specific fragments of DNA polymerase Ⅰ gene （polA） of Tp from 165 whole - blood samples of persons with suspected syphilis or non -syphilis. Compared with serology, the semi -nested PCR method was evaluated in the detection of Tp in whole blood. Results Of 165 tests performed, directly compared with serology, semi -nested PCR showed 82.4% agreement, with a sensitivity of 62.7% and a specificity of 95.9%. There were significant differences between the two PCR assays in detection of Tp from whole blood and the semi - nested polA PCR was much more sensitive than the routine polA PCR （ P 〈 0.01 ）. Conclusions The semi - nestedpolA PCR assay is much more sensitive than the routine pelA PCR assay in detecting low numbers of Tp in whole blood samples as a useful addition to serology for the diagnosis of infectious syphilis. For the semi - nested polA PCR assay, the higher sensitivity for detection of Tp in whole blood should be enhanced.

关 键 词：梅毒螺旋体 半巢式PCR polA 全血 

分 类 号：R377.1[医药卫生—病原生物学]