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Application of MALDI-TOF mass spectrometry for the detection of enterotoxins produced by pathogenic strains of the Bacillus cereus group

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Abstract

Enterotoxins produced by different species of the Bacillus cereus group, such as cytotoxin K1 (CytK1) and non-haemolytic enterotoxin (NHE), have been associated with diarrhoeal food poisoning incidents. Detection of CytK1 is not possible with commercial assays while NHE is recognised by an immunological kit (TECRA) that does not specifically target this protein because it is based on polyclonal antibodies. It is evident that the lack of suitable tools for the study of enterotoxins hampers the possibilities for accurate hazard identification and characterisation in microbial food safety risk assessment. We applied matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS) for the detection of CytK1 and NHE produced by pathogenic strains of the B. cereus group using protein digests from 1D gel electrophoresis. Secretion of CytK1 and two of the three components of NHE was confirmed in supernatants of different B. cereus cultures. For each protein, we introduce biomarkers that could be used for the screening of food poisoning or food/environmental isolates that can secrete enterotoxins. For example, tryptic peptides of 2,310.2 and 1,192.5 Da (calculated mass) can be indicators for CytK1 and NheA, respectively, although a simultaneous detection of other enterotoxin-specific peptides is recommended to assure the presence of a toxin in an unknown sample. Comparison of MALDI-TOF/MS with the TECRA kit showed that our methodological strategy performed well and it had the competitive advantage of specifically detecting NheA. Therefore, MALDI-TOF/MS can be successfully incorporated into risk assessment procedures in order to determine the involvement of strains of the B. cereus group in foodborne outbreaks, including the recently described cytK1 producing species, Bacillus cytotoxicus.

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Acknowledgements

We thank Prof. N. Boon for the critical revisions of this manuscript. This work was financially supported by the Belgian Federal Public Service (FOD) of Health, Food Chain Safety and Environment (RT09/2 BACEREUS) and the Special Research Funds of Ghent University (B/09036/02 fund IV1 31/10/2008–31/10/2012). Bart Devreese is indebted to the Belgian Federal Government-Interuniversity Attraction Pole Action P6/19 and a GOA grant from Ghent University Research Funds. Tom Van de Wiele and Andreja Rajkovic are Postdoctoral Fellows belonging to the Fund for Scientific Research (FWO) of Flanders (Belgium). The authors have declared no conflict of interest.

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Authors and Affiliations

  1. Laboratory for Protein Biochemistry and Biomolecular Engineering (L-PROBE), Department of Biochemistry and Microbiology, Ghent University, K.L. Ledeganckstraat 35, 9000, Ghent, Belgium

    Varvara Tsilia, Bart Devreese & Bart Mesuere

  2. Laboratory of Food Microbiology and Food Preservation (LFMFP), Department of Food Safety and Food Quality, Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Varvara Tsilia, Ilse de Baenst, Andreja Rajkovic & Mieke Uyttendaele

  3. Laboratory of Microbial Ecology and Technology (LabMET), Department of Biochemical and Microbial Technology, Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Varvara Tsilia & Tom Van de Wiele

  4. Technology and Food Science Unit, Institute for Agricultural and Fisheries Research (ILVO), Brusselsesteenweg 370, 9090, Melle, Belgium

    Varvara Tsilia & Marc Heyndrickx

  5. Department of Applied Mathematics and Computer Science, Ghent University, Krijgslaan 281, S9, 9000, Ghent, Belgium

    Bart Mesuere

  6. Department of Pathology, Bacteriology and Poultry Diseases, Ghent University, Salisburylaan 133, 9820, Merelbeke, Belgium

    Marc Heyndrickx

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  1. Varvara Tsilia
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  2. Bart Devreese
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  3. Ilse de Baenst
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  6. Mieke Uyttendaele
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  7. Tom Van de Wiele
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Correspondence to Tom Van de Wiele.

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Tsilia, V., Devreese, B., de Baenst, I. et al. Application of MALDI-TOF mass spectrometry for the detection of enterotoxins produced by pathogenic strains of the Bacillus cereus group. Anal Bioanal Chem 404, 1691–1702 (2012). https://doi.org/10.1007/s00216-012-6254-6

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  • DOI: https://doi.org/10.1007/s00216-012-6254-6

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