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Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS
BACKGROUND: Colistin (polymyxin E) and polymixin B are important bactericidal antibiotics used in the treatment of serious infections caused by multi-drug resistant Gram-negative organisms. Transferrable plasmid-mediated colistin resistance, conferred by the product of the mcr-1 gene, has emerged as...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6390366/ https://www.ncbi.nlm.nih.gov/pubmed/30890899 http://dx.doi.org/10.1186/s12014-019-9228-2 |
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author | Wang, Honghui Chen, Yong Strich, Jeffrey R. Drake, Steven K. Youn, Jung-Ho Rosenberg, Avi Z. Gucek, Marjan McGann, Patrick T. Suffredini, Anthony F. Dekker, John P. |
author_facet | Wang, Honghui Chen, Yong Strich, Jeffrey R. Drake, Steven K. Youn, Jung-Ho Rosenberg, Avi Z. Gucek, Marjan McGann, Patrick T. Suffredini, Anthony F. Dekker, John P. |
author_sort | Wang, Honghui |
collection | PubMed |
description | BACKGROUND: Colistin (polymyxin E) and polymixin B are important bactericidal antibiotics used in the treatment of serious infections caused by multi-drug resistant Gram-negative organisms. Transferrable plasmid-mediated colistin resistance, conferred by the product of the mcr-1 gene, has emerged as a global healthcare threat. Consequently, the rapid detection of the MCR-1 protein in clinical bacterial isolates has become increasingly important. We used a genoproteomic approach to identify unique peptides of the MCR-1 protein that could be detected rapidly by liquid chromatography tandem mass spectrometry (LC–MS/MS). METHODS: MCR-1 tryptic peptides that were efficiently ionized and readily detectable were characterized in a set of mcr-1-containing isolates with triple quadrupole LC–MS. Three optimal peptides were selected for the development of a rapid multiple reaction monitoring LC–MS/MS assay for the MCR-1 protein. To investigate the feasibility of rapid detection of the MCR-1 protein in bacterial isolates using this assay, a blinded 99-sample test set was built that included three additional mcr-1-containing clinical isolates tested in triplicate (9 samples) and 90 negative control isolates. RESULTS: All of the mcr-1-containing isolates in the test set were accurately identified with no false positive detections by three independent, blinded operators, yielding an overall performance of 100% sensitivity and specificity for multiple operators. Among the three peptides tested in this study, the best performing was DTFPQLAK. The isolate-to-result time for the assay as implemented is less than 90 min. CONCLUSIONS: This work demonstrates the feasibility of rapid detection of the MCR-1 protein in bacterial isolates by LC–MS/MS. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12014-019-9228-2) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6390366 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-63903662019-03-19 Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS Wang, Honghui Chen, Yong Strich, Jeffrey R. Drake, Steven K. Youn, Jung-Ho Rosenberg, Avi Z. Gucek, Marjan McGann, Patrick T. Suffredini, Anthony F. Dekker, John P. Clin Proteomics Research BACKGROUND: Colistin (polymyxin E) and polymixin B are important bactericidal antibiotics used in the treatment of serious infections caused by multi-drug resistant Gram-negative organisms. Transferrable plasmid-mediated colistin resistance, conferred by the product of the mcr-1 gene, has emerged as a global healthcare threat. Consequently, the rapid detection of the MCR-1 protein in clinical bacterial isolates has become increasingly important. We used a genoproteomic approach to identify unique peptides of the MCR-1 protein that could be detected rapidly by liquid chromatography tandem mass spectrometry (LC–MS/MS). METHODS: MCR-1 tryptic peptides that were efficiently ionized and readily detectable were characterized in a set of mcr-1-containing isolates with triple quadrupole LC–MS. Three optimal peptides were selected for the development of a rapid multiple reaction monitoring LC–MS/MS assay for the MCR-1 protein. To investigate the feasibility of rapid detection of the MCR-1 protein in bacterial isolates using this assay, a blinded 99-sample test set was built that included three additional mcr-1-containing clinical isolates tested in triplicate (9 samples) and 90 negative control isolates. RESULTS: All of the mcr-1-containing isolates in the test set were accurately identified with no false positive detections by three independent, blinded operators, yielding an overall performance of 100% sensitivity and specificity for multiple operators. Among the three peptides tested in this study, the best performing was DTFPQLAK. The isolate-to-result time for the assay as implemented is less than 90 min. CONCLUSIONS: This work demonstrates the feasibility of rapid detection of the MCR-1 protein in bacterial isolates by LC–MS/MS. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12014-019-9228-2) contains supplementary material, which is available to authorized users. BioMed Central 2019-02-26 /pmc/articles/PMC6390366/ /pubmed/30890899 http://dx.doi.org/10.1186/s12014-019-9228-2 Text en © The Author(s) 2019 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 Wang, Honghui Chen, Yong Strich, Jeffrey R. Drake, Steven K. Youn, Jung-Ho Rosenberg, Avi Z. Gucek, Marjan McGann, Patrick T. Suffredini, Anthony F. Dekker, John P. Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
title | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
title_full | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
title_fullStr | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
title_full_unstemmed | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
title_short | Rapid detection of colistin resistance protein MCR-1 by LC–MS/MS |
title_sort | rapid detection of colistin resistance protein mcr-1 by lc–ms/ms |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6390366/ https://www.ncbi.nlm.nih.gov/pubmed/30890899 http://dx.doi.org/10.1186/s12014-019-9228-2 |
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