外周血检测梅毒螺旋体DNA及其分子亚型的初步观察

Detection of polA gene in whole blood samples and preliminary observations with molecular subtyping of Treponema pallidum

目的 探讨外周血标本检测梅毒螺旋体DNA基因分型的可行性,了解梅毒病程与梅毒螺旋体DNA阳性率及基因分型的关系.方法 收集2012年7月至2013年2月北京3家医院性病中心就诊的181例梅毒患者的临床资料,采用PCR方法检测患者外周血梅毒螺旋体特异性基因polA基因;对pol A基因阳性标本采用巢式PCR法扩增酸性重复蛋白质基因arp和重复基因簇基因tpr进行基因分型.结果 181例梅毒患者中有35例polA基因阳性,其中67例初诊非潜伏梅毒的未治疗患者中24例（35.8％）阳性;26例潜伏梅毒患者均阴性;72例复诊患者中7例（9.7％）阳性;16例血清固定患者中4例（25.0％）阳性.进一步基因分型发现35份polA基因阳性标本中14份为14d亚型,10份为14b亚型,7份为14a亚型,4份为13c亚型,其中血清固定的4份标本中3份为14d亚型,1份为14b亚型.结论 外周血检测梅毒螺旋体基因具有可行性,尽管阳性率较低,但是可靠易行,尤其对血清固定患者具有一定意义.

Objective To detect Treponema pallidum （T.pallidum） DNA with polymerase chain reaction （PCR） in whole blood samples of syphilis patients and analyze their features of subgenotypes.Methods The clinical data of patients were collected from July 2012 to February 2013.And polA gene of syphilis was detected by PCR.The arp and tpr genes of polA gene-positive samples were analyzed by the established genotyping system.Statistical analyses were performed to compare different clinical courses and features to examine their correlations.Results The common treponemal gene target （polA） of 35 samples were detected in whole blood by PCR in 181 samples.A total of 24 cases （35.8%） were positive in 67 patients with newly diagnosed non-latent syphilis untreated patients;26 cases of latent syphilis were negative;7 cases （9.7%） were positive in 72 subsequent visit patients;4 cases （25.0%） were positive in 16 patients with sero-resistant.There were 4 subtypes of 14a （n =7）,14b （n =10）,13c （n =4） and 14d （n =14）.Among those positive samples,there were 4 sero-resistant samples of 3 subtypes 14d and 1 subtype 14b.Conclusions The feasibility of peripheral blood is confirmed.Although the positive rate of whole blood detection of T.pallidum gene is low,the method is both simple and reliable for patients with sero-resistant syphilis.