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Journal of Clinical Microbiology
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Bacteriology

Direct Identification of 80 Percent of Bacteria from Blood Culture Bottles by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using a 10-Minute Extraction Protocol

Loïc Simon, Estelle Ughetto, Alice Gaudart, Nicolas Degand, Romain Lotte, Raymond Ruimy
Karen C. Carroll, Editor
Loïc Simon
Laboratoire de Bactériologie, CHU de Nice, Hôpital de l'Archet 2, Nice, France
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Estelle Ughetto
Laboratoire de Bactériologie, CHU de Nice, Hôpital de l'Archet 2, Nice, France
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Alice Gaudart
Laboratoire de Bactériologie, CHU de Nice, Hôpital de l'Archet 2, Nice, France
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Nicolas Degand
Laboratoire de Bactériologie, CHU de Nice, Hôpital de l'Archet 2, Nice, France
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Romain Lotte
Laboratoire de Bactériologie, CHU de Nice, Hôpital de l'Archet 2, Nice, FranceUniversité Côte d’Azur, Nice, FranceINSERM U1065, C3M, Equipe 6 Virulence Microbienne et Signalisation Inflammatoire, Bâtiment Universitaire Archimed, Nice, France
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Raymond Ruimy
Laboratoire de Bactériologie, CHU de Nice, Hôpital de l'Archet 2, Nice, FranceUniversité Côte d’Azur, Nice, FranceINSERM U1065, C3M, Equipe 6 Virulence Microbienne et Signalisation Inflammatoire, Bâtiment Universitaire Archimed, Nice, France
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Karen C. Carroll
Johns Hopkins University School of Medicine
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DOI: 10.1128/JCM.01278-18
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ABSTRACT

Matrix-assisted laser desorption ionization–time of flight mass spectrometry is not widely used to identify bacteria directly from positive blood culture bottles (BCBs) because of overlong protocols. The objective of this work was to develop and evaluate a simple extraction protocol for reliable identification from BCBs. The 10-min protocol was applied over a 5-month period. Direct identifications on day 0 were compared with those obtained from colonies on day 1 [log(score) of ≥2]. We evaluated a range of seven log(score) thresholds on day 0 from 1.4 to 2.0 to find the lower confidence score that provides the higher percentage of direct identifications without loss of accuracy. With a log(score) threshold of ≥1.5 at day 0, our protocol allowed us to identify 80% of bacteria in 632 BCBs (96% of Enterobacteriaceae, 95% of Staphylococcus aureus, 92% of enterococci, and 62% of streptococci). At least one bacterial species of the mixture was identified in 77% of the polymicrobial samples. The rapidity and reliability of the protocol were factors in its adoption for routine use, allowing us to save up to 24 h in identifying 80% of the bacteria in the BCBs and, thus, to supply useful information to adapt antibiotic therapy when necessary. We currently provide reliable daily direct identifications of staphylococci, enterococci, Enterobacteriaceae, Pseudomonas aeruginosa, and beta-hemolytic streptococci.

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Direct Identification of 80 Percent of Bacteria from Blood Culture Bottles by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using a 10-Minute Extraction Protocol
Loïc Simon, Estelle Ughetto, Alice Gaudart, Nicolas Degand, Romain Lotte, Raymond Ruimy
Journal of Clinical Microbiology Jan 2019, 57 (2) e01278-18; DOI: 10.1128/JCM.01278-18

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Direct Identification of 80 Percent of Bacteria from Blood Culture Bottles by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Using a 10-Minute Extraction Protocol
Loïc Simon, Estelle Ughetto, Alice Gaudart, Nicolas Degand, Romain Lotte, Raymond Ruimy
Journal of Clinical Microbiology Jan 2019, 57 (2) e01278-18; DOI: 10.1128/JCM.01278-18
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KEYWORDS

MALDI-TOF
bacteremia
blood culture bottle
bloodstream infection
direct identification of bacteria
extraction protocol

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