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

Varvara Tsilia UGent, Bart Devreese UGent, Ilse de Baenst UGent, Bart Mesuere UGent, Andreja Rajkovic UGent, Mieke Uyttendaele UGent, Tom Van de Wiele UGent and Marc Heyndrickx UGent (2012) ANALYTICAL AND BIOANALYTICAL CHEMISTRY. 404(6-7). p.1691-1702
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.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
Foodborne pathogen, cereus, Enterotoxins, 1D electrophoresis, MALDI-TOF/MS, Biomarkers, EMETIC TOXIN, QUANTITATIVE-ANALYSIS, HEMOLYSIN-BL, THURINGIENSIS, GENES, PCR, CYTK, IMMUNOASSAY, COMPLEX, FAMILY
journal title
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Anal. Bioanal. Chem.
volume
404
issue
6-7
pages
1691 - 1702
Web of Science type
Article
Web of Science id
000309348400009
JCR category
CHEMISTRY, ANALYTICAL
JCR impact factor
3.659 (2012)
JCR rank
9/74 (2012)
JCR quartile
1 (2012)
ISSN
1618-2642
DOI
10.1007/s00216-012-6254-6
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2971424
handle
http://hdl.handle.net/1854/LU-2971424
date created
2012-08-13 16:01:15
date last changed
2013-01-07 15:34:46
@article{2971424,
  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.},
  author       = {Tsilia, Varvara and Devreese, Bart and de Baenst, Ilse and Mesuere, Bart and Rajkovic, Andreja and Uyttendaele, Mieke and Van de Wiele, Tom and Heyndrickx, Marc},
  issn         = {1618-2642},
  journal      = {ANALYTICAL AND BIOANALYTICAL CHEMISTRY},
  keyword      = {Foodborne pathogen,cereus,Enterotoxins,1D electrophoresis,MALDI-TOF/MS,Biomarkers,EMETIC TOXIN,QUANTITATIVE-ANALYSIS,HEMOLYSIN-BL,THURINGIENSIS,GENES,PCR,CYTK,IMMUNOASSAY,COMPLEX,FAMILY},
  language     = {eng},
  number       = {6-7},
  pages        = {1691--1702},
  title        = {Application of MALDI-TOF mass spectrometry for the detection of enterotoxins produced by pathogenic strains of the Bacillus cereus group},
  url          = {http://dx.doi.org/10.1007/s00216-012-6254-6},
  volume       = {404},
  year         = {2012},
}

Chicago
Tsilia, Varvara, Bart Devreese, Ilse de Baenst, Bart Mesuere, Andreja Rajkovic, Mieke Uyttendaele, Tom Van de Wiele, and Marc Heyndrickx. 2012. “Application of MALDI-TOF Mass Spectrometry for the Detection of Enterotoxins Produced by Pathogenic Strains of the Bacillus Cereus Group.” Analytical and Bioanalytical Chemistry 404 (6-7): 1691–1702.
APA
Tsilia, V., Devreese, B., de Baenst, I., Mesuere, B., Rajkovic, A., Uyttendaele, M., Van de Wiele, T., et al. (2012). Application of MALDI-TOF mass spectrometry for the detection of enterotoxins produced by pathogenic strains of the Bacillus cereus group. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 404(6-7), 1691–1702.
Vancouver
1.
Tsilia V, Devreese B, de Baenst I, Mesuere B, Rajkovic A, Uyttendaele M, et al. Application of MALDI-TOF mass spectrometry for the detection of enterotoxins produced by pathogenic strains of the Bacillus cereus group. ANALYTICAL AND BIOANALYTICAL CHEMISTRY. 2012;404(6-7):1691–702.
MLA
Tsilia, Varvara, Bart Devreese, Ilse de Baenst, et al. “Application of MALDI-TOF Mass Spectrometry for the Detection of Enterotoxins Produced by Pathogenic Strains of the Bacillus Cereus Group.” ANALYTICAL AND BIOANALYTICAL CHEMISTRY 404.6-7 (2012): 1691–1702. Print.