Cargando…

Direct assessment of possible mutations in the 23S rRNA gene encoding macrolide resistance in Chlamydia trachomatis

Reports of potential treatment failure have raised particular concerns regarding the efficacy of the single dose azithromycin regimen in the treatment of urogenital and anorectal Chlamydia trachomatis (CT) infections. Several factors have been suggested, including heterotypic resistance. Antimicrobi...

Descripción completa

Detalles Bibliográficos
Autores principales: van Niekerk, J. M., van Loo, I. H. M., Lucchesi, M., Morré, S. A., Hoebe, C. J. P. A., Dukers-Muijrers, N. H. T. M., Wolffs, P. F. G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9089867/
https://www.ncbi.nlm.nih.gov/pubmed/35536784
http://dx.doi.org/10.1371/journal.pone.0265229
_version_ 1784704606486396928
author van Niekerk, J. M.
van Loo, I. H. M.
Lucchesi, M.
Morré, S. A.
Hoebe, C. J. P. A.
Dukers-Muijrers, N. H. T. M.
Wolffs, P. F. G.
author_facet van Niekerk, J. M.
van Loo, I. H. M.
Lucchesi, M.
Morré, S. A.
Hoebe, C. J. P. A.
Dukers-Muijrers, N. H. T. M.
Wolffs, P. F. G.
author_sort van Niekerk, J. M.
collection PubMed
description Reports of potential treatment failure have raised particular concerns regarding the efficacy of the single dose azithromycin regimen in the treatment of urogenital and anorectal Chlamydia trachomatis (CT) infections. Several factors have been suggested, including heterotypic resistance. Antimicrobial susceptibility testing in CT requires cell culture with serial dilutions of antibiotics, which is laborious and for which there is no standardized testing methodology. One method to partly overcome these difficulties would be to use a genotypic resistance assay, however most current available assays do still require prior CT culture. In order to facilitate the assessment of genotypic resistance directly from clinical samples, without the need for prior culture, the aim of this study was to develop a CT specific PCR assay for the assessment of resistance associated mutations (RAMs) in the 23S rRNA gene, and to evaluate a sample of clinical cases in which CT PCR’s remained positive during follow-up despite azithromycin treatment. Neither the in silico analysis nor the analytical specificity testing demonstrated clinically relevant cross-reactivity with other bacterial species. These results in conjunction with the analytical sensitivity demonstrating consistent CT 23S rRNA gene detection in the range of 10e3 IFU/mL, exemplify the assay’s apt performance. Although no known macrolide RAMs were detected in the clinical cases, the described assay allows future culture independent macrolide RAM surveillance in CT, and increases accessibility for other laboratories to engage in screening.
format Online
Article
Text
id pubmed-9089867
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-90898672022-05-11 Direct assessment of possible mutations in the 23S rRNA gene encoding macrolide resistance in Chlamydia trachomatis van Niekerk, J. M. van Loo, I. H. M. Lucchesi, M. Morré, S. A. Hoebe, C. J. P. A. Dukers-Muijrers, N. H. T. M. Wolffs, P. F. G. PLoS One Research Article Reports of potential treatment failure have raised particular concerns regarding the efficacy of the single dose azithromycin regimen in the treatment of urogenital and anorectal Chlamydia trachomatis (CT) infections. Several factors have been suggested, including heterotypic resistance. Antimicrobial susceptibility testing in CT requires cell culture with serial dilutions of antibiotics, which is laborious and for which there is no standardized testing methodology. One method to partly overcome these difficulties would be to use a genotypic resistance assay, however most current available assays do still require prior CT culture. In order to facilitate the assessment of genotypic resistance directly from clinical samples, without the need for prior culture, the aim of this study was to develop a CT specific PCR assay for the assessment of resistance associated mutations (RAMs) in the 23S rRNA gene, and to evaluate a sample of clinical cases in which CT PCR’s remained positive during follow-up despite azithromycin treatment. Neither the in silico analysis nor the analytical specificity testing demonstrated clinically relevant cross-reactivity with other bacterial species. These results in conjunction with the analytical sensitivity demonstrating consistent CT 23S rRNA gene detection in the range of 10e3 IFU/mL, exemplify the assay’s apt performance. Although no known macrolide RAMs were detected in the clinical cases, the described assay allows future culture independent macrolide RAM surveillance in CT, and increases accessibility for other laboratories to engage in screening. Public Library of Science 2022-05-10 /pmc/articles/PMC9089867/ /pubmed/35536784 http://dx.doi.org/10.1371/journal.pone.0265229 Text en © 2022 van Niekerk et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
van Niekerk, J. M.
van Loo, I. H. M.
Lucchesi, M.
Morré, S. A.
Hoebe, C. J. P. A.
Dukers-Muijrers, N. H. T. M.
Wolffs, P. F. G.
Direct assessment of possible mutations in the 23S rRNA gene encoding macrolide resistance in Chlamydia trachomatis
title Direct assessment of possible mutations in the 23S rRNA gene encoding macrolide resistance in Chlamydia trachomatis
title_full Direct assessment of possible mutations in the 23S rRNA gene encoding macrolide resistance in Chlamydia trachomatis
title_fullStr Direct assessment of possible mutations in the 23S rRNA gene encoding macrolide resistance in Chlamydia trachomatis
title_full_unstemmed Direct assessment of possible mutations in the 23S rRNA gene encoding macrolide resistance in Chlamydia trachomatis
title_short Direct assessment of possible mutations in the 23S rRNA gene encoding macrolide resistance in Chlamydia trachomatis
title_sort direct assessment of possible mutations in the 23s rrna gene encoding macrolide resistance in chlamydia trachomatis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9089867/
https://www.ncbi.nlm.nih.gov/pubmed/35536784
http://dx.doi.org/10.1371/journal.pone.0265229
work_keys_str_mv AT vanniekerkjm directassessmentofpossiblemutationsinthe23srrnageneencodingmacrolideresistanceinchlamydiatrachomatis
AT vanlooihm directassessmentofpossiblemutationsinthe23srrnageneencodingmacrolideresistanceinchlamydiatrachomatis
AT lucchesim directassessmentofpossiblemutationsinthe23srrnageneencodingmacrolideresistanceinchlamydiatrachomatis
AT morresa directassessmentofpossiblemutationsinthe23srrnageneencodingmacrolideresistanceinchlamydiatrachomatis
AT hoebecjpa directassessmentofpossiblemutationsinthe23srrnageneencodingmacrolideresistanceinchlamydiatrachomatis
AT dukersmuijrersnhtm directassessmentofpossiblemutationsinthe23srrnageneencodingmacrolideresistanceinchlamydiatrachomatis
AT wolffspfg directassessmentofpossiblemutationsinthe23srrnageneencodingmacrolideresistanceinchlamydiatrachomatis