Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples

Here, we describe the optimization and validation of five quantitative PCR (qPCR) assays by employing the SYBRGreen chemistry paired with melting curve analysis to detect and quantify clinically relevant antimicrobial resistance genes (ARGs) (i.e. ermB, bla(CTXM1-like), bla(CMY-2), qnrA and qnrS) fr...

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Autores principales: Tolosi, Roberta, Carraro, Lisa, Laconi, Andrea, Piccirillo, Alessandra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8563462/
https://www.ncbi.nlm.nih.gov/pubmed/34754761
http://dx.doi.org/10.1016/j.mex.2021.101488
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author Tolosi, Roberta
Carraro, Lisa
Laconi, Andrea
Piccirillo, Alessandra
author_facet Tolosi, Roberta
Carraro, Lisa
Laconi, Andrea
Piccirillo, Alessandra
author_sort Tolosi, Roberta
collection PubMed
description Here, we describe the optimization and validation of five quantitative PCR (qPCR) assays by employing the SYBRGreen chemistry paired with melting curve analysis to detect and quantify clinically relevant antimicrobial resistance genes (ARGs) (i.e. ermB, bla(CTXM1-like), bla(CMY-2), qnrA and qnrS) from environmental samples (i.e. soil and manure). These five protocols accurately detected and quantified the aforementioned ARGs in complex environmental matrices and represent useful tools for both diagnostic and monitoring activities of resistant bacteria and ARGs into the environment.
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spelling pubmed-85634622021-11-08 Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples Tolosi, Roberta Carraro, Lisa Laconi, Andrea Piccirillo, Alessandra MethodsX Protocol Article Here, we describe the optimization and validation of five quantitative PCR (qPCR) assays by employing the SYBRGreen chemistry paired with melting curve analysis to detect and quantify clinically relevant antimicrobial resistance genes (ARGs) (i.e. ermB, bla(CTXM1-like), bla(CMY-2), qnrA and qnrS) from environmental samples (i.e. soil and manure). These five protocols accurately detected and quantified the aforementioned ARGs in complex environmental matrices and represent useful tools for both diagnostic and monitoring activities of resistant bacteria and ARGs into the environment. Elsevier 2021-08-12 /pmc/articles/PMC8563462/ /pubmed/34754761 http://dx.doi.org/10.1016/j.mex.2021.101488 Text en © 2021 The Authors. Published by Elsevier B.V. https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol Article
Tolosi, Roberta
Carraro, Lisa
Laconi, Andrea
Piccirillo, Alessandra
Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples
title Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples
title_full Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples
title_fullStr Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples
title_full_unstemmed Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples
title_short Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples
title_sort optimization of five qpcr protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples
topic Protocol Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8563462/
https://www.ncbi.nlm.nih.gov/pubmed/34754761
http://dx.doi.org/10.1016/j.mex.2021.101488
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