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Intact Staphylococcus Enterotoxin SEB from Culture Supernatant Detected by MALDI-TOF Mass Spectrometry

Routine identification of pathogens by MALDI-TOF MS (matrix-assisted laser desorption ionisation time-of-flight mass spectrometry) is based on the fingerprint of intracellular proteins. This work evaluated the use of MALDI-TOF MS for the identification of extracellular pathogen factors. A Staphyloco...

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Autores principales: Tonacini, Jenna, Stephan, Dario, Vogel, Guido, Avondet, Marc-André, Kalman, Franka, Crovadore, Julien, Lefort, François, Schnyder, Bruno
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6409910/
https://www.ncbi.nlm.nih.gov/pubmed/30744109
http://dx.doi.org/10.3390/toxins11020101
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author Tonacini, Jenna
Stephan, Dario
Vogel, Guido
Avondet, Marc-André
Kalman, Franka
Crovadore, Julien
Lefort, François
Schnyder, Bruno
author_facet Tonacini, Jenna
Stephan, Dario
Vogel, Guido
Avondet, Marc-André
Kalman, Franka
Crovadore, Julien
Lefort, François
Schnyder, Bruno
author_sort Tonacini, Jenna
collection PubMed
description Routine identification of pathogens by MALDI-TOF MS (matrix-assisted laser desorption ionisation time-of-flight mass spectrometry) is based on the fingerprint of intracellular proteins. This work evaluated the use of MALDI-TOF MS for the identification of extracellular pathogen factors. A Staphylococcus aureus isolate from a food contaminant was exponentially grown in liquid cultures. Secreted proteins were collected using methanol– chloroform precipitation and analysed by MALDI-TOF MS. A main peak m/z 28,250 was demonstrated, which was identified as S.aureus enterotoxin type B (SEB) by using the pure authentic SEB reference of 28.2 kDa and by amino acid sequence analysis. SEB was also detected in this intact form following pasteurization and cooking treatments. Further application of the elaborated MALDI-TOF MS protocol resulted in the detection of SEA at m/z 27,032 and SEC at m/z 27,629. In conclusion, a simple sample preparation from S.aureus cultures and an easy-to-perform identification of pathogen factors SE in intact form represents a promising next-generation application of MALDI-TOF MS.
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spelling pubmed-64099102019-04-01 Intact Staphylococcus Enterotoxin SEB from Culture Supernatant Detected by MALDI-TOF Mass Spectrometry Tonacini, Jenna Stephan, Dario Vogel, Guido Avondet, Marc-André Kalman, Franka Crovadore, Julien Lefort, François Schnyder, Bruno Toxins (Basel) Article Routine identification of pathogens by MALDI-TOF MS (matrix-assisted laser desorption ionisation time-of-flight mass spectrometry) is based on the fingerprint of intracellular proteins. This work evaluated the use of MALDI-TOF MS for the identification of extracellular pathogen factors. A Staphylococcus aureus isolate from a food contaminant was exponentially grown in liquid cultures. Secreted proteins were collected using methanol– chloroform precipitation and analysed by MALDI-TOF MS. A main peak m/z 28,250 was demonstrated, which was identified as S.aureus enterotoxin type B (SEB) by using the pure authentic SEB reference of 28.2 kDa and by amino acid sequence analysis. SEB was also detected in this intact form following pasteurization and cooking treatments. Further application of the elaborated MALDI-TOF MS protocol resulted in the detection of SEA at m/z 27,032 and SEC at m/z 27,629. In conclusion, a simple sample preparation from S.aureus cultures and an easy-to-perform identification of pathogen factors SE in intact form represents a promising next-generation application of MALDI-TOF MS. MDPI 2019-02-09 /pmc/articles/PMC6409910/ /pubmed/30744109 http://dx.doi.org/10.3390/toxins11020101 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Tonacini, Jenna
Stephan, Dario
Vogel, Guido
Avondet, Marc-André
Kalman, Franka
Crovadore, Julien
Lefort, François
Schnyder, Bruno
Intact Staphylococcus Enterotoxin SEB from Culture Supernatant Detected by MALDI-TOF Mass Spectrometry
title Intact Staphylococcus Enterotoxin SEB from Culture Supernatant Detected by MALDI-TOF Mass Spectrometry
title_full Intact Staphylococcus Enterotoxin SEB from Culture Supernatant Detected by MALDI-TOF Mass Spectrometry
title_fullStr Intact Staphylococcus Enterotoxin SEB from Culture Supernatant Detected by MALDI-TOF Mass Spectrometry
title_full_unstemmed Intact Staphylococcus Enterotoxin SEB from Culture Supernatant Detected by MALDI-TOF Mass Spectrometry
title_short Intact Staphylococcus Enterotoxin SEB from Culture Supernatant Detected by MALDI-TOF Mass Spectrometry
title_sort intact staphylococcus enterotoxin seb from culture supernatant detected by maldi-tof mass spectrometry
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6409910/
https://www.ncbi.nlm.nih.gov/pubmed/30744109
http://dx.doi.org/10.3390/toxins11020101
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