High-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in Peruvian isolates

BACKGROUND: The emergence of multidrug-resistant strains is a major health problem especially for countries with high TB incidence such as Peru. In this study, we evaluated High Resolution Melting (HRM) assay in Peruvian isolates for the detection of mutations within rpoB, katG genes and promoter re...

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Autores principales: Galarza, Marco, Fasabi, Manuel, Levano, Kelly S., Castillo, Edith, Barreda, Nadia, Rodriguez, Mitzi, Guio, Heinner
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4899911/
https://www.ncbi.nlm.nih.gov/pubmed/27278526
http://dx.doi.org/10.1186/s12879-016-1615-y
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author Galarza, Marco
Fasabi, Manuel
Levano, Kelly S.
Castillo, Edith
Barreda, Nadia
Rodriguez, Mitzi
Guio, Heinner
author_facet Galarza, Marco
Fasabi, Manuel
Levano, Kelly S.
Castillo, Edith
Barreda, Nadia
Rodriguez, Mitzi
Guio, Heinner
author_sort Galarza, Marco
collection PubMed
description BACKGROUND: The emergence of multidrug-resistant strains is a major health problem especially for countries with high TB incidence such as Peru. In this study, we evaluated High Resolution Melting (HRM) assay in Peruvian isolates for the detection of mutations within rpoB, katG genes and promoter region inhA to determine isoniazid and rifampicin resistance in Mycobacterium tuberculosis (Mtb). METHODS: DNA samples extracted from a total of 167 clinical isolates of Mtb, 89 drug-sensitive and 78 multidrug-resistant, were blindly analyzed by HRM analysis and verified by DNA sequencing. RESULTS: The HRM analysis generated patterns that were specific to distinguish between sensitive and resistance isolates. The sensitivity and specificity of the HRM assays in comparison with drug susceptibility testing (DST) for detection of rifampicin resistance were 98.7 % and 97.5 %, and for isoniazid resistance were 98.7 % and 100 %. CONCLUSION: This study suggests that HRM Analysis could help with rapid diagnosis of MDR-TB cases in Peru.
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spelling pubmed-48999112016-06-13 High-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in Peruvian isolates Galarza, Marco Fasabi, Manuel Levano, Kelly S. Castillo, Edith Barreda, Nadia Rodriguez, Mitzi Guio, Heinner BMC Infect Dis Research Article BACKGROUND: The emergence of multidrug-resistant strains is a major health problem especially for countries with high TB incidence such as Peru. In this study, we evaluated High Resolution Melting (HRM) assay in Peruvian isolates for the detection of mutations within rpoB, katG genes and promoter region inhA to determine isoniazid and rifampicin resistance in Mycobacterium tuberculosis (Mtb). METHODS: DNA samples extracted from a total of 167 clinical isolates of Mtb, 89 drug-sensitive and 78 multidrug-resistant, were blindly analyzed by HRM analysis and verified by DNA sequencing. RESULTS: The HRM analysis generated patterns that were specific to distinguish between sensitive and resistance isolates. The sensitivity and specificity of the HRM assays in comparison with drug susceptibility testing (DST) for detection of rifampicin resistance were 98.7 % and 97.5 %, and for isoniazid resistance were 98.7 % and 100 %. CONCLUSION: This study suggests that HRM Analysis could help with rapid diagnosis of MDR-TB cases in Peru. BioMed Central 2016-06-09 /pmc/articles/PMC4899911/ /pubmed/27278526 http://dx.doi.org/10.1186/s12879-016-1615-y Text en © The Author(s). 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Galarza, Marco
Fasabi, Manuel
Levano, Kelly S.
Castillo, Edith
Barreda, Nadia
Rodriguez, Mitzi
Guio, Heinner
High-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in Peruvian isolates
title High-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in Peruvian isolates
title_full High-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in Peruvian isolates
title_fullStr High-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in Peruvian isolates
title_full_unstemmed High-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in Peruvian isolates
title_short High-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in Peruvian isolates
title_sort high-resolution melting analysis for molecular detection of multidrug resistance tuberculosis in peruvian isolates
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4899911/
https://www.ncbi.nlm.nih.gov/pubmed/27278526
http://dx.doi.org/10.1186/s12879-016-1615-y
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