Evaluation of a PCR Test for Detection of Treponema pallidum in Swabs and Blood
Evaluation of a PCR Test for Detection of Treponema pallidum in Swabs and Blood
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DOI：
10.1128/jcm.00702-11
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发表时间：
2012-03-01
影响因子：
9.4
通讯作者：
Dupin, N.
中科院分区：
文献类型：
--
作者：
Grange, P. A.;Gressier, L.;Dupin, N.
梅毒的诊断是基于梅毒亚种的临床观察、血清学分析和暗视野显微镜(DFM)检测。梅毒是皮肤溃疡中梅毒的病原体。我们用套式聚合酶链式反应(NPCR)从拭子和血液标本中特异性地扩增出梅毒螺旋体的tpp47基因。我们研究了294名疑似梅毒患者和35名健康志愿者的队列。294例患者中一期梅毒87例，二期梅毒103例，潜伏梅毒40例，未检出梅毒。拭子标本梅毒检测结果与梅毒诊断符合率为82%，特异度为95%。NPCR法和DFM法的结果符合得很好(kappa=0.53)。检测外周血单个核细胞(PBMC)、血浆、血清和全血部分的敏感性分别为29%、18%、14.7%和24%，特异性分别为96%、92%、93%和97%。艾滋病毒状况不影响在任何被测试样本中检测到梅毒螺旋体的频率。粘膜或皮肤病变的拭子样本似乎比血液更有助于有效地检测梅毒螺旋体基因组，从而诊断梅毒。
Syphilis diagnosis is based on clinical observation, serological analysis, and dark-field microscopy (DFM) detection of Treponema pallidum subsp. pallidum, the etiological agent of syphilis, in skin ulcers. We performed a nested PCR (nPCR) assay specifically amplifying the tpp47 gene of T. pallidum from swab and blood specimens. We studied a cohort of 294 patients with suspected syphilis and 35 healthy volunteers. Eighty-seven of the 294 patients had primary syphilis, 103 had secondary syphilis, 40 had latent syphilis, and 64 were found not to have syphilis. The T. pallidum nPCR results for swab specimens were highly concordant with syphilis diagnosis, with a sensitivity of 82% and a specificity of 95%. Reasonable agreement was observed between the results obtained with the nPCR and DFM methods (kappa = 0.53). No agreement was found between the nPCR detection of T. pallidum in blood and the diagnosis of syphilis, with sensitivities of 29, 18, 14.7, and 24% and specificities of 96, 92, 93, and 97% for peripheral blood mononuclear cell (PBMC), plasma, serum, and whole-blood fractions, respectively. HIV status did not affect the frequency of T. pallidum detection in any of the specimens tested. Swab specimens from mucosal or skin lesions seemed to be more useful than blood for the efficient detection of the T. pallidum genome and, thus, for the diagnosis of syphilis.