Martin Irene E, Gu Weiming, Yang Yang, Tsang Raymond S W
Division of Pathogenic Neisseria, Syphilis Diagnostics, Vaccine Preventable Bacterial Diseases, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Clin Infect Dis. 2009 Aug 15;49(4):515-21. doi: 10.1086/600878.
The resurgence of syphilis in China requires laboratories to update their methods for molecular epidemiology investigation and surveillance. This study focuses on implementing polymerase chain reaction (PCR) diagnostic tests for syphilis and for detection of molecular subtypes and macrolide resistance among strains causing primary syphilis in the city of Shanghai, China.
Swabs were obtained from the genital lesions of 39 patients who presented with symptoms compatible with primary syphilis. Eight of the patients also provided whole blood samples. Swabs were also obtained from 10 patients without syphilis who presented with genital ulcers. PCR tests to amplify 3 common but unlinked treponemal genes were performed on DNA samples extracted from these specimens. Molecular subtyping was based on genetic characterization of 2 treponemal repeat genes, arp and tpr. Detection of macrolide resistance was accomplished by identification of the 23S ribosomal RNA gene mutation associated with the resistance pattern.
Thirty-eight patients with primary syphilis were found to have Treponema pallidum DNA in their genital lesions by PCR assays using primers that target the bmp, tpp-47, and polA genes. None of the patients without syphilis had positive PCR results. Five molecular subtypes were identified, with 1 type (14f) causing 79% of the cases. All 38 patients were found to be infected with macrolide-resistant strains.
Three common treponemal gene targets (bmp, tpp-47, and polA) were detectable by PCR in patients with primary syphilis. A single clone characterized as 14f and showing macrolide resistance appeared to have caused most of the primary syphilis cases in this study.
梅毒在中国的再度流行要求实验室更新其分子流行病学调查和监测方法。本研究聚焦于在中国上海市实施针对梅毒的聚合酶链反应(PCR)诊断检测,以及对引起一期梅毒的菌株进行分子亚型检测和大环内酯耐药性检测。
从39例出现与一期梅毒相符症状的患者的生殖器病变处采集拭子。其中8例患者还提供了全血样本。另外,从10例患有生殖器溃疡但无梅毒的患者处采集拭子。对从这些标本中提取的DNA样本进行PCR检测,以扩增3个常见但不连锁的密螺旋体基因。分子亚型分型基于两个密螺旋体重复基因arp和tpr的基因特征。通过鉴定与耐药模式相关的23S核糖体RNA基因突变来检测大环内酯耐药性。
使用靶向bmp、tpp - 47和polA基因的引物进行PCR检测,发现38例一期梅毒患者的生殖器病变中有梅毒螺旋体DNA。所有无梅毒的患者PCR结果均为阴性。鉴定出5种分子亚型,其中1种类型(14f)占病例的79%。发现所有38例患者均感染了大环内酯耐药菌株。
在一期梅毒患者中,通过PCR可检测到3个常见的密螺旋体基因靶点(bmp、tpp - 47和polA)。在本研究中,一种被鉴定为14f且表现出大环内酯耐药性的单一克隆似乎导致了大多数一期梅毒病例。