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Accurate Detection of the Four Most Prevalent Carbapenemases in E. coli and K. pneumoniae by High-Resolution Mass Spectrometry
BACKGROUND: At present, phenotypic growth inhibition techniques are used in routine diagnostic microbiology to determine antimicrobial resistance of bacteria. Molecular techniques such as PCR are often used for confirmation but are indirect as they detect particular resistance genes. A direct techni...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2019
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6901907/ https://www.ncbi.nlm.nih.gov/pubmed/31849899 http://dx.doi.org/10.3389/fmicb.2019.02760 |
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author | Foudraine, Dimard E. Dekker, Lennard J. M. Strepis, Nikolaos Bexkens, Michiel L. Klaassen, Corné H. W. Luider, Theo M. Goessens, Wil H. F. |
author_facet | Foudraine, Dimard E. Dekker, Lennard J. M. Strepis, Nikolaos Bexkens, Michiel L. Klaassen, Corné H. W. Luider, Theo M. Goessens, Wil H. F. |
author_sort | Foudraine, Dimard E. |
collection | PubMed |
description | BACKGROUND: At present, phenotypic growth inhibition techniques are used in routine diagnostic microbiology to determine antimicrobial resistance of bacteria. Molecular techniques such as PCR are often used for confirmation but are indirect as they detect particular resistance genes. A direct technique would be able to detect the proteins of the resistance mechanism itself. In the present study targeted high resolution mass spectrometry assay was developed for the simultaneous detection of KPC, OXA-48-like, NDM, and VIM carbapenemases. METHODS: Carbapenemase specific target peptides were defined by comparing available sequences in GenBank. Selected peptide sequences were validated using 62 Klebsiella pneumoniae and Escherichia coli isolates containing: 16 KPC, 21 OXA-48-like, 16 NDM, 13 VIM genes, and 21 carbapenemase negative isolates. RESULTS: For each carbapenemase, two candidate peptides were validated. Method validation was performed in a blinded manner for all 83 isolates. All carbapenemases were detected. The majority was detected by both target peptides. All target peptides were 100% specific in the tested isolates and no peptide carry-over was detected. CONCLUSION: The applied targeted bottom-up mass spectrometry technique is able to accurately detect the four most prevalent carbapenemases in a single analysis. |
format | Online Article Text |
id | pubmed-6901907 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-69019072019-12-17 Accurate Detection of the Four Most Prevalent Carbapenemases in E. coli and K. pneumoniae by High-Resolution Mass Spectrometry Foudraine, Dimard E. Dekker, Lennard J. M. Strepis, Nikolaos Bexkens, Michiel L. Klaassen, Corné H. W. Luider, Theo M. Goessens, Wil H. F. Front Microbiol Microbiology BACKGROUND: At present, phenotypic growth inhibition techniques are used in routine diagnostic microbiology to determine antimicrobial resistance of bacteria. Molecular techniques such as PCR are often used for confirmation but are indirect as they detect particular resistance genes. A direct technique would be able to detect the proteins of the resistance mechanism itself. In the present study targeted high resolution mass spectrometry assay was developed for the simultaneous detection of KPC, OXA-48-like, NDM, and VIM carbapenemases. METHODS: Carbapenemase specific target peptides were defined by comparing available sequences in GenBank. Selected peptide sequences were validated using 62 Klebsiella pneumoniae and Escherichia coli isolates containing: 16 KPC, 21 OXA-48-like, 16 NDM, 13 VIM genes, and 21 carbapenemase negative isolates. RESULTS: For each carbapenemase, two candidate peptides were validated. Method validation was performed in a blinded manner for all 83 isolates. All carbapenemases were detected. The majority was detected by both target peptides. All target peptides were 100% specific in the tested isolates and no peptide carry-over was detected. CONCLUSION: The applied targeted bottom-up mass spectrometry technique is able to accurately detect the four most prevalent carbapenemases in a single analysis. Frontiers Media S.A. 2019-11-26 /pmc/articles/PMC6901907/ /pubmed/31849899 http://dx.doi.org/10.3389/fmicb.2019.02760 Text en Copyright © 2019 Foudraine, Dekker, Strepis, Bexkens, Klaassen, Luider and Goessens. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Foudraine, Dimard E. Dekker, Lennard J. M. Strepis, Nikolaos Bexkens, Michiel L. Klaassen, Corné H. W. Luider, Theo M. Goessens, Wil H. F. Accurate Detection of the Four Most Prevalent Carbapenemases in E. coli and K. pneumoniae by High-Resolution Mass Spectrometry |
title | Accurate Detection of the Four Most Prevalent Carbapenemases in E. coli and K. pneumoniae by High-Resolution Mass Spectrometry |
title_full | Accurate Detection of the Four Most Prevalent Carbapenemases in E. coli and K. pneumoniae by High-Resolution Mass Spectrometry |
title_fullStr | Accurate Detection of the Four Most Prevalent Carbapenemases in E. coli and K. pneumoniae by High-Resolution Mass Spectrometry |
title_full_unstemmed | Accurate Detection of the Four Most Prevalent Carbapenemases in E. coli and K. pneumoniae by High-Resolution Mass Spectrometry |
title_short | Accurate Detection of the Four Most Prevalent Carbapenemases in E. coli and K. pneumoniae by High-Resolution Mass Spectrometry |
title_sort | accurate detection of the four most prevalent carbapenemases in e. coli and k. pneumoniae by high-resolution mass spectrometry |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6901907/ https://www.ncbi.nlm.nih.gov/pubmed/31849899 http://dx.doi.org/10.3389/fmicb.2019.02760 |
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