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

Chen, Cheng‐Yen; Chi, Kai-Hua; Pillay, Allan; Nachamkin, Eli; Su, John R.; Ballard, Ronald C.

doi:10.1128/jcm.02770-12

Cited by 57 publications

(40 citation statements)

References 28 publications

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“…Subsequent investigation showed it was present in 14.6% of T. pallidum samples from that country (29). It was not detected in South African or Taiwanese T. pallidum specimens (18,19), but it has been found at relatively low levels (10 to 13.2%) in the United States (12,30) and in 5.6% of samples in London, United Kingdom (14). Of note in these studies, no samples were simultaneously positive for both A2058G and A2059G.…”

Section: Discussionmentioning

confidence: 95%

Azithromycin-Resistant Syphilis-Causing Strains in Sydney, Australia: Prevalence and Risk Factors

et al. 2014

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e Azithromycin has shown high efficacy in randomized trials when used for treating infectious syphilis in Africa. However, its use in clinical practice has been limited by the development of antimicrobial drug resistance. Resistance has not previously been reported from Australasia. The aim of this study was to determine the prevalence of and risk factors for azithromycin-resistant syphilis-causing strains in Sydney, Australia. We evaluated 409 samples that were PCR positive for Treponema pallidum DNA collected between 2004 and 2011 for the presence of the A2058G mutation, which confers resistance to macrolide antibiotics such as azithromycin. Overall, 84% of samples harbored the mutation. The prevalence of the mutation increased during the study period (P trend, 0.003). We also collected clinical and demographic data on 220 patients from whom these samples had been collected to determine factors associated with the A2058G mutation; 97% were from men who have sex with men. Reporting sex in countries other than Australia was associated with less macrolide resistance (adjusted odds ratio, 0.25; 95% confidence interval, 0.09 to 0.66; P ‫؍‬ 0.005), with other study factors showing no association (age, HIV status, recent macrolide use, stage of syphilis, or history of prior syphilis). Azithromycin cannot be recommended as an alternative treatment for syphilis in Sydney.

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Section: Discussionmentioning

confidence: 95%

Azithromycin-Resistant Syphilis-Causing Strains in Sydney, Australia: Prevalence and Risk Factors

et al. 2014

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“…In addition, a previously used panel of nonpathogenic treponemes (T. denticola, T. refringens, and T. phagedenis) and other microorganisms was included to determine the specificity of the assay. 12 The stability of the tp0858 PCR target was also tested with the real-time quadriplex PCR using DNA from the TPpertenue strain CDC-1 obtained from multiple rabbit passages performed sequentially seven times in 1986 and three times in 1987.…”

Section: Methodsmentioning

confidence: 99%

Molecular Differentiation of Treponema pallidum Subspecies in Skin Ulceration Clinically Suspected as Yaws in Vanuatu Using Real-Time Multiplex PCR and Serological Methods

Chi¹,

Danavall²,

Taleo³

et al. 2015

The American Society of Tropical Medicine and Hygiene

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Abstract. We developed a TaqMan-based real-time quadriplex polymerase chain reaction (PCR) to simultaneously detect Treponema pallidum subspecies pallidum, T. pallidum subsp. pertenue, and T. pallidum subsp. endemicum, the causative agents of venereal syphilis, yaws, and bejel, respectively. The PCR assay was applied to samples from skin ulcerations of clinically presumptive yaws cases among children on Tanna Island, Vanuatu. Another real-time triplex PCR was used to screen for the point mutations in the 23S rRNA genes that have previously been associated with azithromycin resistance in T. pallidum subsp. pallidum strains. Seropositivity by the classical syphilis serological tests was 35.5% among children with skin ulcerations clinically suspected with yaws, whereas the presence of T. pallidum subsp. pertenue DNA was only found in lesions from 15.5% of children. No evidence of T. pallidum subsp. pertenue infection, by either PCR or serology was found in~59% of cases indicating alternative causes of yaws-like lesions in this endemic area.

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“…As this approach is time-consuming and labor-intensive, development of a faster (but equally sensitive and specific) test that can help assess macrolide susceptibility among T. pallidum strains is definitely desirable, even though these assays will still need to be performed in selected reference laboratories. Toward this end, a real-time PCR assay using fluorescence resonance energy transfer (FRET) probes and melting curve analysis was developed for one of the two known point mutations (223), while a real-time multiplex PCR assay for detection of both mutations has been recently published by Chen et al (224).…”

Section: Direct Detection Methodsmentioning

confidence: 99%

The Endemic Treponematoses

2014

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SUMMARY The agents of human treponematoses include four closely related members of the genus Treponema : three subspecies of Treponema pallidum plus Treponema carateum . T. pallidum subsp. pallidum causes venereal syphilis, while T. pallidum subsp. pertenue , T. pallidum subsp. endemicum , and T. carateum are the agents of the endemic treponematoses yaws, bejel (or endemic syphilis), and pinta, respectively. All human treponematoses share remarkable similarities in pathogenesis and clinical manifestations, consistent with the high genetic and antigenic relatedness of their etiological agents. Distinctive features have been identified in terms of age of acquisition, most common mode of transmission, and capacity for invasion of the central nervous system and fetus, although the accuracy of these purported differences is debated among investigators and no biological basis for these differences has been identified to date. In 2012, the World Health Organization (WHO) officially set a goal for yaws eradication by 2020. This challenging but potentially feasible endeavor is favored by the adoption of oral azithromycin for mass treatment and the currently focused distribution of yaws and endemic treponematoses and has revived global interest in these fascinating diseases and their causative agents.

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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

Cited by 57 publications

References 28 publications

Azithromycin-Resistant Syphilis-Causing Strains in Sydney, Australia: Prevalence and Risk Factors

Azithromycin-Resistant Syphilis-Causing Strains in Sydney, Australia: Prevalence and Risk Factors

Molecular Differentiation of Treponema pallidum Subspecies in Skin Ulceration Clinically Suspected as Yaws in Vanuatu Using Real-Time Multiplex PCR and Serological Methods

The Endemic Treponematoses

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