Cargando…
High-Resolution Melting Analysis for the Rapid Detection of Fluoroquinolone and Streptomycin Resistance in Mycobacterium tuberculosis
BACKGROUND: Molecular methods for the detection of drug-resistant tuberculosis are potentially more rapid than conventional culture-based drug susceptibility testing, facilitating the commencement of appropriate treatment for patients with drug resistant tuberculosis. We aimed to develop and evaluat...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3283711/ https://www.ncbi.nlm.nih.gov/pubmed/22363772 http://dx.doi.org/10.1371/journal.pone.0031934 |
_version_ | 1782224241813356544 |
---|---|
author | Lee, Ann S. G. Ong, Danny C. T. Wong, Joshua C. L. Siu, Gilman K. H. Yam, Wing-Cheong |
author_facet | Lee, Ann S. G. Ong, Danny C. T. Wong, Joshua C. L. Siu, Gilman K. H. Yam, Wing-Cheong |
author_sort | Lee, Ann S. G. |
collection | PubMed |
description | BACKGROUND: Molecular methods for the detection of drug-resistant tuberculosis are potentially more rapid than conventional culture-based drug susceptibility testing, facilitating the commencement of appropriate treatment for patients with drug resistant tuberculosis. We aimed to develop and evaluate high-resolution melting (HRM) assays for the detection of mutations within gyrA, rpsL, and rrs, for the determination of fluoroquinolone and streptomycin resistance in Mycobacterium tuberculosis (MTB). METHODOLOGY/PRINCIPAL FINDINGS: A blinded series of DNA samples extracted from a total of 92 clinical isolates of MTB were analyzed by HRM analysis, and the results were verified using DNA sequencing. The sensitivity and specificity of the HRM assays in comparison with drug susceptibility testing were 74.1% and 100.0% for the detection of fluoroquinolone resistance, and 87.5% and 100.0% for streptomycin resistance. Five isolates with low level resistance to ofloxacin had no mutations detected in gyrA, possibly due to the action of efflux pumps, or false negativity due to mixed infections. One fluoroquinolone-resistant isolate had a mutation in a region of gyrA not encompassed by our assay. Six streptomycin-resistant strains had undetectable mutations by HRM and DNA sequencing, which may be explained by the fact that not all streptomycin-resistant isolates have mutations within rpsL and rrs, and suggesting that other targets may be involved. CONCLUSION: The HRM assays described here are potentially useful adjunct tests for the efficient determination of fluoroquinolone and streptomycin resistance in MTB, and could facilitate the timely administration of appropriate treatment for patients infected with drug-resistant TB. |
format | Online Article Text |
id | pubmed-3283711 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-32837112012-02-23 High-Resolution Melting Analysis for the Rapid Detection of Fluoroquinolone and Streptomycin Resistance in Mycobacterium tuberculosis Lee, Ann S. G. Ong, Danny C. T. Wong, Joshua C. L. Siu, Gilman K. H. Yam, Wing-Cheong PLoS One Research Article BACKGROUND: Molecular methods for the detection of drug-resistant tuberculosis are potentially more rapid than conventional culture-based drug susceptibility testing, facilitating the commencement of appropriate treatment for patients with drug resistant tuberculosis. We aimed to develop and evaluate high-resolution melting (HRM) assays for the detection of mutations within gyrA, rpsL, and rrs, for the determination of fluoroquinolone and streptomycin resistance in Mycobacterium tuberculosis (MTB). METHODOLOGY/PRINCIPAL FINDINGS: A blinded series of DNA samples extracted from a total of 92 clinical isolates of MTB were analyzed by HRM analysis, and the results were verified using DNA sequencing. The sensitivity and specificity of the HRM assays in comparison with drug susceptibility testing were 74.1% and 100.0% for the detection of fluoroquinolone resistance, and 87.5% and 100.0% for streptomycin resistance. Five isolates with low level resistance to ofloxacin had no mutations detected in gyrA, possibly due to the action of efflux pumps, or false negativity due to mixed infections. One fluoroquinolone-resistant isolate had a mutation in a region of gyrA not encompassed by our assay. Six streptomycin-resistant strains had undetectable mutations by HRM and DNA sequencing, which may be explained by the fact that not all streptomycin-resistant isolates have mutations within rpsL and rrs, and suggesting that other targets may be involved. CONCLUSION: The HRM assays described here are potentially useful adjunct tests for the efficient determination of fluoroquinolone and streptomycin resistance in MTB, and could facilitate the timely administration of appropriate treatment for patients infected with drug-resistant TB. Public Library of Science 2012-02-21 /pmc/articles/PMC3283711/ /pubmed/22363772 http://dx.doi.org/10.1371/journal.pone.0031934 Text en Lee et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Lee, Ann S. G. Ong, Danny C. T. Wong, Joshua C. L. Siu, Gilman K. H. Yam, Wing-Cheong High-Resolution Melting Analysis for the Rapid Detection of Fluoroquinolone and Streptomycin Resistance in Mycobacterium tuberculosis |
title | High-Resolution Melting Analysis for the Rapid Detection of Fluoroquinolone and Streptomycin Resistance in Mycobacterium tuberculosis
|
title_full | High-Resolution Melting Analysis for the Rapid Detection of Fluoroquinolone and Streptomycin Resistance in Mycobacterium tuberculosis
|
title_fullStr | High-Resolution Melting Analysis for the Rapid Detection of Fluoroquinolone and Streptomycin Resistance in Mycobacterium tuberculosis
|
title_full_unstemmed | High-Resolution Melting Analysis for the Rapid Detection of Fluoroquinolone and Streptomycin Resistance in Mycobacterium tuberculosis
|
title_short | High-Resolution Melting Analysis for the Rapid Detection of Fluoroquinolone and Streptomycin Resistance in Mycobacterium tuberculosis
|
title_sort | high-resolution melting analysis for the rapid detection of fluoroquinolone and streptomycin resistance in mycobacterium tuberculosis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3283711/ https://www.ncbi.nlm.nih.gov/pubmed/22363772 http://dx.doi.org/10.1371/journal.pone.0031934 |
work_keys_str_mv | AT leeannsg highresolutionmeltinganalysisfortherapiddetectionoffluoroquinoloneandstreptomycinresistanceinmycobacteriumtuberculosis AT ongdannyct highresolutionmeltinganalysisfortherapiddetectionoffluoroquinoloneandstreptomycinresistanceinmycobacteriumtuberculosis AT wongjoshuacl highresolutionmeltinganalysisfortherapiddetectionoffluoroquinoloneandstreptomycinresistanceinmycobacteriumtuberculosis AT siugilmankh highresolutionmeltinganalysisfortherapiddetectionoffluoroquinoloneandstreptomycinresistanceinmycobacteriumtuberculosis AT yamwingcheong highresolutionmeltinganalysisfortherapiddetectionoffluoroquinoloneandstreptomycinresistanceinmycobacteriumtuberculosis |