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Direct Detection of KPC Peak from Positive Blood Cultures Using MALDI-TOF MS: Are We There Yet?

Detecting carbapenemase-associated carbapenem resistance is a subject of major clinical and epidemiological concern as it influences therapeutic choice. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed as a means to assess bacterial resist...

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Autores principales: Moreira, Natália Kehl, Wilhelm, Camila Mörschbächer, Echevarria, Aymê Duarte, Volpato, Fabiana Caroline Zempulski, Wink, Priscila Lamb, Barth, Afonso Luís, Caierão, Juliana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10045339/
https://www.ncbi.nlm.nih.gov/pubmed/36978468
http://dx.doi.org/10.3390/antibiotics12030601
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author Moreira, Natália Kehl
Wilhelm, Camila Mörschbächer
Echevarria, Aymê Duarte
Volpato, Fabiana Caroline Zempulski
Wink, Priscila Lamb
Barth, Afonso Luís
Caierão, Juliana
author_facet Moreira, Natália Kehl
Wilhelm, Camila Mörschbächer
Echevarria, Aymê Duarte
Volpato, Fabiana Caroline Zempulski
Wink, Priscila Lamb
Barth, Afonso Luís
Caierão, Juliana
author_sort Moreira, Natália Kehl
collection PubMed
description Detecting carbapenemase-associated carbapenem resistance is a subject of major clinical and epidemiological concern as it influences therapeutic choice. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed as a means to assess bacterial resistance mechanisms. We aimed to detect the KPC enzyme directly from positive blood cultures using MALDI-TOF MS. To do so, 102 clinical Enterobacteria were evaluated, including 59 bla(KPC) positives. Proteins were extracted using formic acid, isopropyl alcohol, and water (17:33:50) and spotted onto a steel target plate using the double-layer sinapinic acid technique. Two parameters were considered: (i) the visual detection of a clear peak with the expected KPC m/z and (ii) the evaluation of the relative intensity of the ions in the peak. A peak was observed in 56/59 bla(KPC)-positive isolates (94.9% sensitivity), with no false-positive results (100% specificity). When considering intensity, with a cut-off ≥120 (a.u.), sensitivity was 94.9% and specificity was 95.3%. We proposed a “buffer” zone, with intermediate values of intensity (115 to 125) reaching 100% sensitivity and specificity. The detection of KPC peaks directly from positive blood cultures using MALDI-TOF MS is feasible and rapid, which may improve appropriate patient therapy and antimicrobial stewardship.
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spelling pubmed-100453392023-03-29 Direct Detection of KPC Peak from Positive Blood Cultures Using MALDI-TOF MS: Are We There Yet? Moreira, Natália Kehl Wilhelm, Camila Mörschbächer Echevarria, Aymê Duarte Volpato, Fabiana Caroline Zempulski Wink, Priscila Lamb Barth, Afonso Luís Caierão, Juliana Antibiotics (Basel) Article Detecting carbapenemase-associated carbapenem resistance is a subject of major clinical and epidemiological concern as it influences therapeutic choice. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed as a means to assess bacterial resistance mechanisms. We aimed to detect the KPC enzyme directly from positive blood cultures using MALDI-TOF MS. To do so, 102 clinical Enterobacteria were evaluated, including 59 bla(KPC) positives. Proteins were extracted using formic acid, isopropyl alcohol, and water (17:33:50) and spotted onto a steel target plate using the double-layer sinapinic acid technique. Two parameters were considered: (i) the visual detection of a clear peak with the expected KPC m/z and (ii) the evaluation of the relative intensity of the ions in the peak. A peak was observed in 56/59 bla(KPC)-positive isolates (94.9% sensitivity), with no false-positive results (100% specificity). When considering intensity, with a cut-off ≥120 (a.u.), sensitivity was 94.9% and specificity was 95.3%. We proposed a “buffer” zone, with intermediate values of intensity (115 to 125) reaching 100% sensitivity and specificity. The detection of KPC peaks directly from positive blood cultures using MALDI-TOF MS is feasible and rapid, which may improve appropriate patient therapy and antimicrobial stewardship. MDPI 2023-03-17 /pmc/articles/PMC10045339/ /pubmed/36978468 http://dx.doi.org/10.3390/antibiotics12030601 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Moreira, Natália Kehl
Wilhelm, Camila Mörschbächer
Echevarria, Aymê Duarte
Volpato, Fabiana Caroline Zempulski
Wink, Priscila Lamb
Barth, Afonso Luís
Caierão, Juliana
Direct Detection of KPC Peak from Positive Blood Cultures Using MALDI-TOF MS: Are We There Yet?
title Direct Detection of KPC Peak from Positive Blood Cultures Using MALDI-TOF MS: Are We There Yet?
title_full Direct Detection of KPC Peak from Positive Blood Cultures Using MALDI-TOF MS: Are We There Yet?
title_fullStr Direct Detection of KPC Peak from Positive Blood Cultures Using MALDI-TOF MS: Are We There Yet?
title_full_unstemmed Direct Detection of KPC Peak from Positive Blood Cultures Using MALDI-TOF MS: Are We There Yet?
title_short Direct Detection of KPC Peak from Positive Blood Cultures Using MALDI-TOF MS: Are We There Yet?
title_sort direct detection of kpc peak from positive blood cultures using maldi-tof ms: are we there yet?
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10045339/
https://www.ncbi.nlm.nih.gov/pubmed/36978468
http://dx.doi.org/10.3390/antibiotics12030601
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