The establishment of a duplex real-time PCR assay for rapid and simultaneous detection of bla(NDM) and bla(KPC) genes in bacteria

The latest threat of multidrug-resistant Gram-negative bacteria corresponds to the emergence of carbapenemase New Delhi metallo-β-lactamase (NDM) and Klebsiella pneumoniae carbapenemase (KPC) producers. Rapid molecular detection is essential to limit their spread. In this study, a duplex real-time p...

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Autores principales: Zheng, Fen, Sun, Jingjing, Cheng, Cancan, Rui, Yongyu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Research
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3816589/
https://www.ncbi.nlm.nih.gov/pubmed/24143953
http://dx.doi.org/10.1186/1476-0711-12-30
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author Zheng, Fen
Sun, Jingjing
Cheng, Cancan
Rui, Yongyu
author_facet Zheng, Fen
Sun, Jingjing
Cheng, Cancan
Rui, Yongyu
author_sort Zheng, Fen
collection PubMed
description The latest threat of multidrug-resistant Gram-negative bacteria corresponds to the emergence of carbapenemase New Delhi metallo-β-lactamase (NDM) and Klebsiella pneumoniae carbapenemase (KPC) producers. Rapid molecular detection is essential to limit their spread. In this study, a duplex real-time polymerase chain reaction (PCR) that was specific for the detection of bla(NDM) and bla(KPC) with the same limit of detection of ten plasmid copies was developed. The assay was linear over eight log dilutions for bla(NDM) (R(2) = 0.971; slope, -3.273) and bla(KPC) (R(2) = 0.992; slope, -2.997) with efficiencies of 102% and 115%, respectively. The assay was validated with 157 clinical isolates and showed 100% concordance with conventional PCR. The excellent performance of the duplex PCR assay makes it a powerful tool for surveillance of the carbapenemases NDM and KPC.
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spelling pubmed-38165892013-11-05 The establishment of a duplex real-time PCR assay for rapid and simultaneous detection of bla(NDM) and bla(KPC) genes in bacteria Zheng, Fen Sun, Jingjing Cheng, Cancan Rui, Yongyu Ann Clin Microbiol Antimicrob Research The latest threat of multidrug-resistant Gram-negative bacteria corresponds to the emergence of carbapenemase New Delhi metallo-β-lactamase (NDM) and Klebsiella pneumoniae carbapenemase (KPC) producers. Rapid molecular detection is essential to limit their spread. In this study, a duplex real-time polymerase chain reaction (PCR) that was specific for the detection of bla(NDM) and bla(KPC) with the same limit of detection of ten plasmid copies was developed. The assay was linear over eight log dilutions for bla(NDM) (R(2) = 0.971; slope, -3.273) and bla(KPC) (R(2) = 0.992; slope, -2.997) with efficiencies of 102% and 115%, respectively. The assay was validated with 157 clinical isolates and showed 100% concordance with conventional PCR. The excellent performance of the duplex PCR assay makes it a powerful tool for surveillance of the carbapenemases NDM and KPC. BioMed Central 2013-10-22 /pmc/articles/PMC3816589/ /pubmed/24143953 http://dx.doi.org/10.1186/1476-0711-12-30 Text en Copyright © 2013 Zheng et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 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
Zheng, Fen
Sun, Jingjing
Cheng, Cancan
Rui, Yongyu
The establishment of a duplex real-time PCR assay for rapid and simultaneous detection of bla(NDM) and bla(KPC) genes in bacteria
title The establishment of a duplex real-time PCR assay for rapid and simultaneous detection of bla(NDM) and bla(KPC) genes in bacteria
title_full The establishment of a duplex real-time PCR assay for rapid and simultaneous detection of bla(NDM) and bla(KPC) genes in bacteria
title_fullStr The establishment of a duplex real-time PCR assay for rapid and simultaneous detection of bla(NDM) and bla(KPC) genes in bacteria
title_full_unstemmed The establishment of a duplex real-time PCR assay for rapid and simultaneous detection of bla(NDM) and bla(KPC) genes in bacteria
title_short The establishment of a duplex real-time PCR assay for rapid and simultaneous detection of bla(NDM) and bla(KPC) genes in bacteria
title_sort establishment of a duplex real-time pcr assay for rapid and simultaneous detection of bla(ndm) and bla(kpc) genes in bacteria
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3816589/
https://www.ncbi.nlm.nih.gov/pubmed/24143953
http://dx.doi.org/10.1186/1476-0711-12-30
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