Rapid species diagnosis for invasive candidiasis using mass spectrometry.

BACKGROUND: Matrix-assisted laser desorption ionisation time of flight mass spectrometry (MALDI TOF-MS) allows the identification of most bacteria and an increasing number of fungi. The potential for the highest clinical benefit of such methods would be in severe acute infections that require prompt...

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Main Authors: Carine Marinach-Patrice, Arnaud Fekkar, Ralitsa Atanasova, Johanna Gomes, Laura Djamdjian, Jean-Yves Brossas, Isabelle Meyer, Pierre Buffet, Georges Snounou, Annick Datry, Christophe Hennequin, Jean-Louis Golmard, Dominique Mazier
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2010-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC2810333?pdf=render
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author Carine Marinach-Patrice
Arnaud Fekkar
Ralitsa Atanasova
Johanna Gomes
Laura Djamdjian
Jean-Yves Brossas
Isabelle Meyer
Pierre Buffet
Georges Snounou
Annick Datry
Christophe Hennequin
Jean-Louis Golmard
Dominique Mazier
author_facet Carine Marinach-Patrice
Arnaud Fekkar
Ralitsa Atanasova
Johanna Gomes
Laura Djamdjian
Jean-Yves Brossas
Isabelle Meyer
Pierre Buffet
Georges Snounou
Annick Datry
Christophe Hennequin
Jean-Louis Golmard
Dominique Mazier
author_sort Carine Marinach-Patrice
collection DOAJ
description BACKGROUND: Matrix-assisted laser desorption ionisation time of flight mass spectrometry (MALDI TOF-MS) allows the identification of most bacteria and an increasing number of fungi. The potential for the highest clinical benefit of such methods would be in severe acute infections that require prompt treatment adapted to the infecting species. Our objective was to determine whether yeasts could be identified directly from a positive blood culture, avoiding the 1-3 days subculture step currently required before any therapeutic adjustments can be made. METHODOLOGY/PRINCIPAL FINDINGS: Using human blood spiked with Candida albicans to simulate blood cultures, we optimized protocols to obtain MALDI TOF-MS fingerprints where signals from blood proteins are reduced. Simulated cultures elaborated using a set of 12 strains belonging to 6 different species were then tested. Quantifiable spectral differences in the 5000-7400 Da mass range allowed to discriminate between these species and to build a reference database. The validation of the method and the statistical approach to spectral analysis were conducted using individual simulated blood cultures of 36 additional strains (six for each species). Correct identification of the species of these strains was obtained. CONCLUSIONS/SIGNIFICANCE: Direct MALDI TOF-MS analysis of aliquots from positive blood cultures allowed rapid and accurate identification of the main Candida species, thus obviating the need for sub-culturing on specific media. Subsequent to this proof-of-principle demonstration, the method can be extended to other clinically relevant yeast species, and applied to an adequate number of clinical samples in order to establish its potential to improve antimicrobial management of patients with fungemia.
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spelling doaj.art-a92522fb68654e34a04b935bf4c6a4152022-12-22T01:59:26ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-01-0151e886210.1371/journal.pone.0008862Rapid species diagnosis for invasive candidiasis using mass spectrometry.Carine Marinach-PatriceArnaud FekkarRalitsa AtanasovaJohanna GomesLaura DjamdjianJean-Yves BrossasIsabelle MeyerPierre BuffetGeorges SnounouAnnick DatryChristophe HennequinJean-Louis GolmardDominique MazierBACKGROUND: Matrix-assisted laser desorption ionisation time of flight mass spectrometry (MALDI TOF-MS) allows the identification of most bacteria and an increasing number of fungi. The potential for the highest clinical benefit of such methods would be in severe acute infections that require prompt treatment adapted to the infecting species. Our objective was to determine whether yeasts could be identified directly from a positive blood culture, avoiding the 1-3 days subculture step currently required before any therapeutic adjustments can be made. METHODOLOGY/PRINCIPAL FINDINGS: Using human blood spiked with Candida albicans to simulate blood cultures, we optimized protocols to obtain MALDI TOF-MS fingerprints where signals from blood proteins are reduced. Simulated cultures elaborated using a set of 12 strains belonging to 6 different species were then tested. Quantifiable spectral differences in the 5000-7400 Da mass range allowed to discriminate between these species and to build a reference database. The validation of the method and the statistical approach to spectral analysis were conducted using individual simulated blood cultures of 36 additional strains (six for each species). Correct identification of the species of these strains was obtained. CONCLUSIONS/SIGNIFICANCE: Direct MALDI TOF-MS analysis of aliquots from positive blood cultures allowed rapid and accurate identification of the main Candida species, thus obviating the need for sub-culturing on specific media. Subsequent to this proof-of-principle demonstration, the method can be extended to other clinically relevant yeast species, and applied to an adequate number of clinical samples in order to establish its potential to improve antimicrobial management of patients with fungemia.http://europepmc.org/articles/PMC2810333?pdf=render
spellingShingle Carine Marinach-Patrice
Arnaud Fekkar
Ralitsa Atanasova
Johanna Gomes
Laura Djamdjian
Jean-Yves Brossas
Isabelle Meyer
Pierre Buffet
Georges Snounou
Annick Datry
Christophe Hennequin
Jean-Louis Golmard
Dominique Mazier
Rapid species diagnosis for invasive candidiasis using mass spectrometry.
PLoS ONE
title Rapid species diagnosis for invasive candidiasis using mass spectrometry.
title_full Rapid species diagnosis for invasive candidiasis using mass spectrometry.
title_fullStr Rapid species diagnosis for invasive candidiasis using mass spectrometry.
title_full_unstemmed Rapid species diagnosis for invasive candidiasis using mass spectrometry.
title_short Rapid species diagnosis for invasive candidiasis using mass spectrometry.
title_sort rapid species diagnosis for invasive candidiasis using mass spectrometry
url http://europepmc.org/articles/PMC2810333?pdf=render
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