Molecular Subtyping of Treponema pallidum subsp pallidum in Lisbon, Portugal
Molecular Subtyping of Treponema pallidum subsp pallidum in Lisbon, Portugal
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DOI：
10.1128/jcm.00287-08
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发表时间：
2009-08-01
影响因子：
9.4
通讯作者：
Pereira, F. Martins
中科院分区：
文献类型：
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作者：
Castro, R.;Prieto, E.;Pereira, F. Martins
本研究的目的是评价梅毒螺旋体分型的分子方法的重复性。pallidum，并区分该微生物菌株与梅毒患者菌株。我们研究了82例不同阶段梅毒患者的212份标本（14例一期梅毒，7例二期梅毒和61例潜伏梅毒）。标本分布如下：生殖器溃疡（n = 9）、皮肤和粘膜病变（n = 7）、血液（n = 82）、血浆（n = 82）和耳垂刮片（n = 32）。通过PCR技术分析样品以扩增聚合酶基因I（polA）的片段。根据tpr和阿普两个可变基因的分析，对阳性样本进行分型。polA阳性的90个样本中有62个产生了可分型的梅毒螺旋体DNA。所有皮肤病变中，T。发现苍白球（6/6 [100%]）含有足够的DNA用于生物体分型。还可以对7/9（77.7%）生殖器溃疡样本、13/22（59.1%）血液样本、20/32（62.5%）血浆样本和16/21（76.2%）耳垂刮片进行DNA分型。在来自同一患者的所有样本中鉴定出相同的亚型。鉴定了5种分子亚型（亚型10a、14 a、14 c、14 f和14 g），其中最常见的亚型是亚型14 a，最不常见的亚型是亚型10a。总之，本研究中使用的分型技术似乎具有良好的重复性。据我们所知，10a亚型是首次鉴定。需要进一步的研究来解释这种亚型在葡萄牙的存在，即它与在讲葡萄牙语的非洲国家流行的梅毒螺旋体菌株的关系。
The objectives of this study were to evaluate the reproducibility of a molecular method for the subtyping of Treponema pallidum subsp. pallidum and to discriminate strains of this microorganism from strains from patients with syphilis. We studied 212 specimens from a total of 82 patients with different stages of syphilis (14 primary, 7 secondary and 61 latent syphilis). The specimens were distributed as follows: genital ulcers (n = 9), skin and mucosal lesions (n = 7), blood (n = 82), plasma (n = 82), and ear lobe scrapings (n = 32). The samples were assayed by a PCR technique to amplify a segment of the polymerase gene I (polA). Positive samples were typed on the basis of the analysis of two variable genes, tpr and arp. Sixty-two of the 90 samples positive for polA yielded typeable Treponema pallidum DNA. All skin lesions in which T. pallidum was identified (six of six [100%]) were found to contain enough DNA for typing of the organism. It was also possible to type DNA from 7/9 (77.7%) genital ulcer samples, 13/22 (59.1%) blood samples, 20/32 (62.5%) plasma samples, and 16/21 (76.2%) ear lobe scrapings. The same subtype was identified in all samples from the same patient. Five molecular subtypes (subtypes 10a, 14a, 14c, 14f, and 14g) were identified, with the most frequently found subtype being subtype 14a and the least frequently found subtype being subtype 10a. In conclusion, the subtyping technique used in this study seems to have good reproducibility. To our knowledge, subtype 10a was identified for the first time. Further studies are needed to explain the presence of this subtype in Portugal, namely, its relationship to the Treponema pallidum strains circulating in the African countries where Portuguese is spoken.