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

Abstract

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.

Fig. 1. The detection rate of T. pallidum DNA in blood samples from all patients by Tpp47-Tp-PCR and polA-Tp-PCR.

a The positive specimens of polA-Tp-PCR and/or Tpp47-Tp-PCR among all patients (n = 262). b The positive specimens of polA-Tp-PCR and/or Tpp47-Tp-PCR in the primary syphilis patients (n = 84). c The positive specimens of polA-Tp-PCR and/or Tpp47-Tp-PCR in the secondary syphilis patients (n = 97). d The positive specimens of polA-Tp-PCR and/or Tpp47-Tp-PCR in the latent syphilis patients (n = 81)