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Metabolomic Investigation of Staphylococcus aureus Antibiotic Susceptibility by Liquid Chromatography Coupled to High-Resolution Mass Spectrometry

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Functional Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1871))

Abstract

Staphylococcus aureus is a major human pathogen that can readily acquire antibiotic resistance. For instance, methicillin-resistant S. aureus represents a major cause of hospital- and community-acquired bacterial infections. In this chapter, we first provide a detailed protocol for obtaining unbiased and reproducible S. aureus metabolic profiles. The resulting intracellular metabolome is then analyzed in an untargeted manner by using both hydrophilic interaction liquid chromatography and pentafluorophenyl-propyl columns coupled to high-resolution mass spectrometry. Such analyses are done in conjunction with our in-house spectral database to identify with high confidence as many meaningful S. aureus metabolites as possible. Under these conditions, we can routinely monitor more than 200 annotated S. aureus metabolites. We also indicate how this protocol can be used to investigate the metabolic differences between methicillin-resistant and susceptible strains.

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Acknowledgements

This work was supported by bioMérieux S.A. and the Association Nationale de la Recherche et de la Technologie (ANRT). S.A.-C. is the recipient of a CIFRE fellowship (grant number 2011/1474). This work was also supported by the Commissariat à l’Energie Atomique et aux Energies Alternatives and the MetaboHUB infrastructure (ANR-11-INBS-0010 grant).

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Authors and Affiliations

  1. Service de Pharmacologie et d’Immunoanalyse, Laboratoire d’Etude du Métabolisme des Médicaments, CEA, INRA, Université Paris Saclay, MetaboHUB, Gif-sur-Yvette, France

    Sandrine Aros-Calt, Florence A. Castelli, Patricia Lamourette, Christophe Junot & François Fenaille

  2. bioMérieux S.A., Marcy l’Etoile, France

    Sandrine Aros-Calt, Gaspard Gervasi & Bruno H. Muller

Authors
  1. Sandrine Aros-Calt
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  2. Florence A. Castelli
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  3. Patricia Lamourette
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  4. Gaspard Gervasi
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  5. Christophe Junot
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  6. Bruno H. Muller
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  7. François Fenaille
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Corresponding author

Correspondence to François Fenaille .

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Editors and Affiliations

  1. Array Bridge Inc., St. Louis, MO, USA

    Xing Wang

  2. Biotech Support Group LLC, Monmouth Junction, NJ, USA

    Matthew Kuruc

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Aros-Calt, S. et al. (2019). Metabolomic Investigation of Staphylococcus aureus Antibiotic Susceptibility by Liquid Chromatography Coupled to High-Resolution Mass Spectrometry. In: Wang, X., Kuruc, M. (eds) Functional Proteomics. Methods in Molecular Biology, vol 1871. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8814-3_18

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  • DOI: https://doi.org/10.1007/978-1-4939-8814-3_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8813-6

  • Online ISBN: 978-1-4939-8814-3

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