Detection of the A2058G and A2059G 23S rRNA Gene Point Mutations Associated with Azithromycin Resistance in Treponema pallidum by Use of a TaqMan Real-Time Multiplex PCR Assay
TLDR
The TaqMan-based real-time triplex PCR assay offers an alternative to conventional nested PCR and restriction fragment length polymorphism analyses for the rapid detection of both point mutations associated with macrolide resistance in T. pallidum.Abstract:
Macrolide treatment failure in syphilis patients is associated with a single point mutation (either A2058G or A2059G) in both copies of the 23S rRNA gene in Treponema pallidum strains. The conventional method for the detection of both point mutations uses nested PCR combined with restriction enzyme digestions, which is laborious and time-consuming. We initially developed a TaqMan-based real-time duplex PCR assay for detection of the A2058G mutation, and upon discovery of the A2059G mutation, we modified the assay into a triplex format to simultaneously detect both mutations. The point mutations detected by the real-time triplex PCR were confirmed by pyrosequencing. A total of 129 specimens PCR positive for T. pallidum that were obtained from an azithromycin resistance surveillance study conducted in the United States were analyzed. Sixty-six (51.2%) of the 129 samples with the A2058G mutation were identified by both real-time PCR assays. Of the remaining 63 samples that were identified as having a macrolide-susceptible genotype by the duplex PCR assay, 17 (27%) were found to contain the A2059G mutation by the triplex PCR. The proportions of macrolide-susceptible versus -resistant genotypes harboring either the A2058G or the A2059G mutation among the T. pallidum strains were 35.6, 51.2, and 13.2%, respectively. None of the T. pallidum strains examined had both point mutations. The TaqMan-based real-time triplex PCR assay offers an alternative to conventional nested PCR and restriction fragment length polymorphism analyses for the rapid detection of both point mutations associated with macrolide resistance in T. pallidum.read more
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Journal ArticleDOI
The Endemic Treponematoses
TL;DR: All human treponematoses share remarkable similarities in pathogenesis and clinical manifestations, consistent with the high genetic and antigenic relatedness of their etiological agents.
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Syphilis: Re-emergence of an old foe.
TL;DR: Genomic and proteomic studies have provided some new information concerning how this spirochete may evade host defense mechanisms to persist for long periods in the host.
Journal ArticleDOI
Syphilis: antibiotic treatment and resistance.
TL;DR: Information is presented here on the diagnosis and recommended antibiotic treatment of syphilis and the challenge of macrolide-resistant T. pallidum.
Journal ArticleDOI
A Cross-Sectional Study of ‘Yaws’ in Districts of Ghana Which Have Previously Undertaken Azithromycin Mass Drug Administration for Trachoma Control
Rosanna Ghinai,Philip El-Duah,Kai Hua Chi,Allan Pillay,Anthony W. Solomon,Robin L. Bailey,Nsiire Agana,David Mabey,Cheng-Yen Chen,Yaw Adu-Sarkodie,Michael Marks +10 more
TL;DR: A cross-sectional survey in thirty communities in two districts of Ghana, previously treated for trachoma, found no evidence of ongoing transmission of yaws and integration of diagnostic testing into both pre and post-MDA surveillance systems is required to better inform yaws control programmes.
Journal ArticleDOI
Molecular differentiation of Treponema pallidum subspecies in skin ulceration clinically suspected as yaws in Vanuatu using real-time multiplex PCR and serological methods.
Kai-Hua Chi,Damien Danavall,Fasihah Taleo,Allan Pillay,Tun Ye,Eli Nachamkin,Jacob L. Kool,David Fegan,Kingsley Asiedu,Lasse S. Vestergaard,Ronald C. Ballard,Cheng-Yen Chen +11 more
TL;DR: A TaqMan-based real-time quadriplex polymerase chain reaction (PCR) to simultaneously detect Treponema pallidum subspecies pallidums, T. pertenue, and T. pallidUM subsp.
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