Peptide Markers for Rapid Detection of KPC Carbapenemase by LC-MS/MS

Abstract Carbapenemase producing organisms (CPOs) represent an urgent public health threat, and the need for new rapid methods to detect these organisms has been widely recognized. CPOs carrying the Klebsiella pneumoniae carbapenemase (bla KPC ) gene have caused outbreaks globally with substantial a...

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Autores principales:	Honghui Wang, Steven K. Drake, Jung-Ho Youn, Avi Z. Rosenberg, Yong Chen, Marjan Gucek, Anthony F. Suffredini, John P. Dekker
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0818e8dae94e4d92bda5bd76772b5d1e
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spelling	oai:doaj.org-article:0818e8dae94e4d92bda5bd76772b5d1e2021-12-02T11:53:09ZPeptide Markers for Rapid Detection of KPC Carbapenemase by LC-MS/MS10.1038/s41598-017-02749-22045-2322https://doaj.org/article/0818e8dae94e4d92bda5bd76772b5d1e2017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02749-2https://doaj.org/toc/2045-2322Abstract Carbapenemase producing organisms (CPOs) represent an urgent public health threat, and the need for new rapid methods to detect these organisms has been widely recognized. CPOs carrying the Klebsiella pneumoniae carbapenemase (bla KPC ) gene have caused outbreaks globally with substantial attributable mortality. Here we describe the validation of a rapid MS method for the direct detection of unique tryptic peptides of the KPC protein in clinical bacterial isolates with an isolate-to-result time of less than 90 minutes. Using a genoproteomic discovery approach that combines theoretical peptidome analysis and liquid chromatography-tandem MS (LC-MS/MS), we selected three high abundance peptide markers of the KPC protein that can be robustly detected following rapid tryptic digestion. Protein BLAST analysis confirmed that the chosen peptide markers were unique to KPC. A blinded validation set containing 20 KPC-positive and 80 KPC-negative clinical isolates, performed in triplicate (300 runs) demonstrated 100% sensitivity and 100% specificity (60/60 positive identifications, 240/240 negative identifications) using defined rules for positive calls. The most robust tryptic peptide marker in the validation was LTLGSALAAPQR. The peptide discovery and detection methods validated here are general and should be broadly applicable to allow the direct and rapid detection of other resistance determinants.Honghui WangSteven K. DrakeJung-Ho YounAvi Z. RosenbergYong ChenMarjan GucekAnthony F. SuffrediniJohn P. DekkerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Honghui Wang Steven K. Drake Jung-Ho Youn Avi Z. Rosenberg Yong Chen Marjan Gucek Anthony F. Suffredini John P. Dekker Peptide Markers for Rapid Detection of KPC Carbapenemase by LC-MS/MS
description	Abstract Carbapenemase producing organisms (CPOs) represent an urgent public health threat, and the need for new rapid methods to detect these organisms has been widely recognized. CPOs carrying the Klebsiella pneumoniae carbapenemase (bla KPC ) gene have caused outbreaks globally with substantial attributable mortality. Here we describe the validation of a rapid MS method for the direct detection of unique tryptic peptides of the KPC protein in clinical bacterial isolates with an isolate-to-result time of less than 90 minutes. Using a genoproteomic discovery approach that combines theoretical peptidome analysis and liquid chromatography-tandem MS (LC-MS/MS), we selected three high abundance peptide markers of the KPC protein that can be robustly detected following rapid tryptic digestion. Protein BLAST analysis confirmed that the chosen peptide markers were unique to KPC. A blinded validation set containing 20 KPC-positive and 80 KPC-negative clinical isolates, performed in triplicate (300 runs) demonstrated 100% sensitivity and 100% specificity (60/60 positive identifications, 240/240 negative identifications) using defined rules for positive calls. The most robust tryptic peptide marker in the validation was LTLGSALAAPQR. The peptide discovery and detection methods validated here are general and should be broadly applicable to allow the direct and rapid detection of other resistance determinants.
format	article
author	Honghui Wang Steven K. Drake Jung-Ho Youn Avi Z. Rosenberg Yong Chen Marjan Gucek Anthony F. Suffredini John P. Dekker
author_facet	Honghui Wang Steven K. Drake Jung-Ho Youn Avi Z. Rosenberg Yong Chen Marjan Gucek Anthony F. Suffredini John P. Dekker
author_sort	Honghui Wang
title	Peptide Markers for Rapid Detection of KPC Carbapenemase by LC-MS/MS
title_short	Peptide Markers for Rapid Detection of KPC Carbapenemase by LC-MS/MS
title_full	Peptide Markers for Rapid Detection of KPC Carbapenemase by LC-MS/MS
title_fullStr	Peptide Markers for Rapid Detection of KPC Carbapenemase by LC-MS/MS
title_full_unstemmed	Peptide Markers for Rapid Detection of KPC Carbapenemase by LC-MS/MS
title_sort	peptide markers for rapid detection of kpc carbapenemase by lc-ms/ms
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/0818e8dae94e4d92bda5bd76772b5d1e
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Peptide Markers for Rapid Detection of KPC Carbapenemase by LC-MS/MS

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