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Evaluation of MALDI-ToF Mass Spectrometry for Rapid Detection of Cereulide from Bacillus cereus Cultures

Joerg Doellinger, Andy Schneider, Timo Stark, Monika Ehling-Schulz, View ORCID ProfilePeter Lasch
doi: https://doi.org/10.1101/869958
Joerg Doellinger
1Proteomics and Spectroscopy unit (ZBS 6) at the Centre for Biological Threats and Special Pathogens, Robert Koch-Institute, Seestraße 10, D-13353 Berlin, Germany
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  • For correspondence: doellingerj@rki.de
Andy Schneider
1Proteomics and Spectroscopy unit (ZBS 6) at the Centre for Biological Threats and Special Pathogens, Robert Koch-Institute, Seestraße 10, D-13353 Berlin, Germany
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Timo Stark
2Technical University of Munich, Food Chemistry and Molecular Sensory Science, 85354 Freising, Lise-Meitner-Straße 34, Munich, Germany
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Monika Ehling-Schulz
3Functional Microbiology, Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine Vienna, 1210, Vienna, Austria
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Peter Lasch
1Proteomics and Spectroscopy unit (ZBS 6) at the Centre for Biological Threats and Special Pathogens, Robert Koch-Institute, Seestraße 10, D-13353 Berlin, Germany
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  • ORCID record for Peter Lasch
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1. Abstract

Bacillus cereus plays an often unrecognized role in food borne diseases. Food poisoning caused by this pathogen is manifested by either diarrhea or emesis. While different enterotoxins have been linked to the diarrheal type of B. cereus infections, the emetic toxin cereulide is responsible for the second type. Due to the relatively high prevalence of cereulide associated food poisoning, methods for simple and reliable detection of cereulide producing strains are of utmost importance. Currently, liquid-chromatography coupled to mass spectrometry (LC-MS) is used for sensitive, specific and quantitative cereulide detection, but this technique requires specialized LC-MS equipment, which is often not available in microbiology routine diagnostic laboratories.

The last decade has witnessed the advent of matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-ToF MS) as a simple, rapid and cost-efficient technique for identification of microbial pathogens in routine diagnostics. Just recently, two different studies reported on the application of MALDI-ToF MS for either the differentiation of emetic and non-emetic strains of B. cereus or for direct detection of cereulide from bacterial colony smears. However, no method evaluation and optimization was performed in frame of these studies. Thus, additional investigations on the selectivity and sensitivity of MALDI-TOF MS for cereulide detection are needed before implementation of this method in routine diagnostics can be considered. These aspects prompted us to investigate open or controversial issues and to systematically test sample preparation methods, commonly used for microbial identification for their suitability to detect the emetic toxin directly from bacteria.

Based on our experimental findings we propose a MALDI-ToF MS workflow that allows identification of B. cereus and sensitive detection of cereulide in parallel, using standard, linear-mode MALDI-ToF MS equipment. The experimental protocol is based on the well-established ethanol/formic acid extraction method and offers, if required, possibilities for further characterization by more sophisticated LC-MS-based methods. In summary, the ease of use and the achieved level of analytical sensitivity as well as the wide-spread availability of standard MALDI-ToF MS equipment in clinical microbiological laboratories provides a promising tool to improve and to facilitate routine diagnostics of B. cereus associated food intoxications.

  • Abbreviations

    ACN
    acetonitrile
    FA
    formic acid
    HCCA
    α-cyano-4-hydroxycinnamic acid
    LC
    liquid-chromatography
    LDI
    laser desorption/ionization
    LOD
    limit of detection
    MALDI-ToF
    matrix-assisted laser desorption/ionization - time–of–flight
    MS
    mass spectrometry
    NRPS
    non-ribosomal peptide synthetase
    SIDA
    stable isotope dilution analysis
    SNR
    signal-to-noise ratio
    Th
    Thomson
    TFA
    trifluoroacetic acid
  • Copyright 
    The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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    Posted December 10, 2019.
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    Evaluation of MALDI-ToF Mass Spectrometry for Rapid Detection of Cereulide from Bacillus cereus Cultures
    Joerg Doellinger, Andy Schneider, Timo Stark, Monika Ehling-Schulz, Peter Lasch
    bioRxiv 869958; doi: https://doi.org/10.1101/869958
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    Evaluation of MALDI-ToF Mass Spectrometry for Rapid Detection of Cereulide from Bacillus cereus Cultures
    Joerg Doellinger, Andy Schneider, Timo Stark, Monika Ehling-Schulz, Peter Lasch
    bioRxiv 869958; doi: https://doi.org/10.1101/869958

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