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An application of competitive reporter monitored amplification (CMA) for rapid detection of single nucleotide polymorphisms (SNPs)

Single nucleotide polymorphisms (SNPs) are essential parameters in molecular diagnostics and can be used for the early detection and clinical prognosis in various diseases. Available methods for SNP detection are still labor-intensive and require a complex laboratory infrastructure, which are not su...

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Autores principales: Havlicek, Juliane, Rivera-Milla, Eric, Slickers, Peter, Andres, Sönke, Feuerriegel, Silke, Niemann, Stefan, Merker, Matthias, Labugger, Ines
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5574540/
https://www.ncbi.nlm.nih.gov/pubmed/28850612
http://dx.doi.org/10.1371/journal.pone.0183561
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author Havlicek, Juliane
Rivera-Milla, Eric
Slickers, Peter
Andres, Sönke
Feuerriegel, Silke
Niemann, Stefan
Merker, Matthias
Labugger, Ines
author_facet Havlicek, Juliane
Rivera-Milla, Eric
Slickers, Peter
Andres, Sönke
Feuerriegel, Silke
Niemann, Stefan
Merker, Matthias
Labugger, Ines
author_sort Havlicek, Juliane
collection PubMed
description Single nucleotide polymorphisms (SNPs) are essential parameters in molecular diagnostics and can be used for the early detection and clinical prognosis in various diseases. Available methods for SNP detection are still labor-intensive and require a complex laboratory infrastructure, which are not suitable for the usage in resource-limited settings. Thus, there is an urgent need for a simple, reliable and rapid approach. In this paper we modified the previously developed competitive reporter monitored amplification (CMA) technique for the detection of resistance mediating SNPs in Mycobacterium tuberculosis complex (MTBC) strains. As a proof-of-principle for the application of the CMA-based SNP assay in routine molecular tuberculosis diagnostic, we show that the assay recognizes resistance mediating SNPs for rifampicin, isoniazid and ethambutol from either isolated DNA or heat inactivated M. tuberculosis cell cultures. The CMA-based SNP assay can identify the most prevalent resistance mediating mutations in the genes rpoB, katG, embB, and the promotor region of inhA within one hour.
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spelling pubmed-55745402017-09-15 An application of competitive reporter monitored amplification (CMA) for rapid detection of single nucleotide polymorphisms (SNPs) Havlicek, Juliane Rivera-Milla, Eric Slickers, Peter Andres, Sönke Feuerriegel, Silke Niemann, Stefan Merker, Matthias Labugger, Ines PLoS One Research Article Single nucleotide polymorphisms (SNPs) are essential parameters in molecular diagnostics and can be used for the early detection and clinical prognosis in various diseases. Available methods for SNP detection are still labor-intensive and require a complex laboratory infrastructure, which are not suitable for the usage in resource-limited settings. Thus, there is an urgent need for a simple, reliable and rapid approach. In this paper we modified the previously developed competitive reporter monitored amplification (CMA) technique for the detection of resistance mediating SNPs in Mycobacterium tuberculosis complex (MTBC) strains. As a proof-of-principle for the application of the CMA-based SNP assay in routine molecular tuberculosis diagnostic, we show that the assay recognizes resistance mediating SNPs for rifampicin, isoniazid and ethambutol from either isolated DNA or heat inactivated M. tuberculosis cell cultures. The CMA-based SNP assay can identify the most prevalent resistance mediating mutations in the genes rpoB, katG, embB, and the promotor region of inhA within one hour. Public Library of Science 2017-08-29 /pmc/articles/PMC5574540/ /pubmed/28850612 http://dx.doi.org/10.1371/journal.pone.0183561 Text en © 2017 Havlicek 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 (http://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
Havlicek, Juliane
Rivera-Milla, Eric
Slickers, Peter
Andres, Sönke
Feuerriegel, Silke
Niemann, Stefan
Merker, Matthias
Labugger, Ines
An application of competitive reporter monitored amplification (CMA) for rapid detection of single nucleotide polymorphisms (SNPs)
title An application of competitive reporter monitored amplification (CMA) for rapid detection of single nucleotide polymorphisms (SNPs)
title_full An application of competitive reporter monitored amplification (CMA) for rapid detection of single nucleotide polymorphisms (SNPs)
title_fullStr An application of competitive reporter monitored amplification (CMA) for rapid detection of single nucleotide polymorphisms (SNPs)
title_full_unstemmed An application of competitive reporter monitored amplification (CMA) for rapid detection of single nucleotide polymorphisms (SNPs)
title_short An application of competitive reporter monitored amplification (CMA) for rapid detection of single nucleotide polymorphisms (SNPs)
title_sort application of competitive reporter monitored amplification (cma) for rapid detection of single nucleotide polymorphisms (snps)
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5574540/
https://www.ncbi.nlm.nih.gov/pubmed/28850612
http://dx.doi.org/10.1371/journal.pone.0183561
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