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Sensitive detection of Treponema pallidum DNA from the whole blood of patients with syphilis by the nested PCR assay

机译:巢式PCR检测灵敏检测梅毒患者全血中的梅毒螺旋体DNA

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摘要

The aim of this work was to investigate the application of the nested PCR assay for the detection of Treponema pallidum (TP) DNA from the blood of patients with different stages of syphilis. In this study, a nested PCR method targeting the Tpp47 and polA genes (Tpp47-Tp-PCR and polA-Tp-PCR) was developed to detect TP-DNA in whole blood samples collected from 262 patients with different stages of syphilis (84 primary syphilis, 97 secondary syphilis, and 81 latent syphilis patients). The PCR assay detected T. pallidum DNA in 53.6% and 62.9% of the patients with primary and secondary syphilis, respectively, which was much higher than the detection levels in patients with latent syphilis (7.4%) (both p < 0.001). For primary syphilis, a low RPR (0–16) was correlated with a higher detection rate of TP-DNA, whereas for secondary syphilis, the higher detection rate of blood TP-DNA was correlated with higher blood RPR titers (at or beyond 32). For latent syphilis, TP-DNA was only detectable by PCR in the early phase of the latent infection. Thus, blood RPR titers were correlated with the blood T. pallidum burden, but the correlations varied with primary and secondary syphilis. The results indicate that nested PCR is a sensitive method for detecting blood TP-DNA and is especially useful for detecting early syphilis including primary syphilis and secondary syphilis. The findings also suggest that the PCR assay may be used to complement other methods to enhance the diagnosis of syphilis.
机译:这项工作的目的是研究巢式PCR检测法在梅毒不同阶段患者血液中检测梅毒螺旋体(TP)DNA的应用。在这项研究中,开发了一种针对Tpp47和polA基因的巢式PCR方法(Tpp47-Tp-PCR和polA-Tp-PCR),用于检测从262例梅毒患者的不同阶段采集的全血样品中的TP-DNA(84梅毒,97例继发性梅毒和81例潜在梅毒患者)。 PCR检测分别在原发性和继发性梅毒患者中分别检测出梅毒螺旋体DNA的53.6%和62.9%,远远高于潜伏梅毒患者的检出水平(7.4%)(p <0.001)。对于原发性梅毒,低RPR(0-16)与较高的TP-DNA检测率相关,而对于继发性梅毒,对血液TP-DNA的较高检测率与较高的血液RPR滴度相关(≥32) )。对于潜伏梅毒,仅在潜伏感染的早期通过PCR才能检测到TP-DNA。因此,血液RPR滴度与血液梅毒螺旋体负担相关,但相关性随原发性和继发性梅毒而异。结果表明,巢式PCR是检测血液TP-DNA的灵敏方法,特别适用于检测包括梅毒和继发梅毒在内的早期梅毒。研究结果还表明,PCR检测可用于补充其他方法,以增强梅毒的诊断。

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