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Volatile organic compound analysis by ion molecule reaction mass spectrometry for Gram-positive bacteria differentiation

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Abstract

Approximately 50 % of all clinically proven infections in critically ill patients are caused by Gram-positive bacteria. The timely and appropriate treatment of these infections is vital in order to avoid negative outcomes. Hence, fast and reliable methods are needed for the early detection and identification of microorganisms. Recently, direct mass spectrometry-based analysis of volatile organic compounds emitted by microorganisms has been employed to study Gram-negative bacteria. Here, we report a feasibility study of ion molecule reaction mass spectrometry (IMR-MS) for in vitro growth detection and species differentiation of selected Gram-positive bacteria that are frequently isolated in blood culture samples, namely, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, and Staphylococcus epidermidis. Ion molecule reaction mass spectrometry was used to analyze the headspace above cultures containing Gram-positive bacteria incubated at 37 °C starting with 102 colony-forming units (CFU)/ml. Measurements to determine the presence of volatile organic compounds were performed 4, 8, and 24 h after incubation, respectively. The detection of microbial growth was accomplished already after 8 h in cultures containing E. faecalis. After 24 h of incubation, characteristic mass spectra were obtained for all species. Processing these mass spectra by hierarchic clustering and principal component analysis (PCA) enabled us to differentiate between bacterial species. IMR‐MS in conjunction with a cumulative end-point model provides the means for rapid growth detection and differentiation of Gram-positive bacteria on the species level, typically within an analysis time of less than 3 min per sample.

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Acknowledgment

The authors are grateful to Kirsten Weinert for her excellent technical assistance during the study period. Avacta/Oxford Medical Diagnostics Ltd. provided financial support and V&F Medical Development GmbH provided the mass spectrometry system.

Conflict of interest

M.E.D. received an indirect research grant from Avacta/Oxford Medical Diagnostics Ltd. V&F reimbursed the travel expenses of M.E.D. and C.H. to the ASA annual meetings and the German Anaesthesia Congress 2006–2010. S.P. is a scientist employed by V&F. J.V. is the owner of V&F. W.D. is a scientist employed by Oxford Medical Diagnostics Ltd. C.K., G.S., and S.S. have no conflict of interest to declare.

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Authors and Affiliations

  1. Department of Anesthesiology, University Hospital Großhadern, Ludwig-Maximilians-University of Munich, 81366, Munich, Germany

    M. E. Dolch, C. Hornuss, C. Klocke & G. Schelling

  2. V&F Medical Development GmbH, Andreas-Hofer-Str. 15, Absam, Austria

    S. Praun & J. Villinger

  3. Avacta/Oxford Medical Diagnostics Ltd., Centre for Innovation & Enterprise, Oxford University Begbroke Science Park, Oxford, UK

    W. Denzer

  4. Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, Ludwig-Maximilians-University of Munich, Munich, Germany

    S. Schubert

Authors
  1. M. E. Dolch
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  2. C. Hornuss
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  3. C. Klocke
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  4. S. Praun
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  5. J. Villinger
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  6. W. Denzer
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  7. G. Schelling
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  8. S. Schubert
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Correspondence to M. E. Dolch.

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Dolch, M.E., Hornuss, C., Klocke, C. et al. Volatile organic compound analysis by ion molecule reaction mass spectrometry for Gram-positive bacteria differentiation. Eur J Clin Microbiol Infect Dis 31, 3007–3013 (2012). https://doi.org/10.1007/s10096-012-1654-2

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  • DOI: https://doi.org/10.1007/s10096-012-1654-2

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