Differentiation of Cronobacter spp. by tryptic digestion of the cell suspension followed by MALDI-TOF MS analysis

Intact cell MALDI-TOF mass spectrometry is a rapid tool for the identification and classification of microorganisms, now widely used even in clinical laboratories. However, its distinctive power is not sufficient for some closely-related species. The genus Cronobacter, formerly known as Enterobacter...

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Bibliographic Details
Published in:Journal of microbiological methods 2014-03, Vol.98, p.105-113
Main Authors: Krásný, Lukáš, Rohlová, Eva, Růžičková, Helena, Šantrůček, Jiří, Hynek, Radovan, Hochel, Igor
Format: Article
Language:English
Subjects:
Bacterial Typing Techniques - methods
Bacteriological Techniques - methods
Biotyper
Cronobacter
Cronobacter - isolation & purification
Cronobacter sakazakii - isolation & purification
Enterobacter sakazakii
MALDI-TOF
Mass spectrometry
Polymerase Chain Reaction - methods
Software
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
Trypsin
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Summary:Intact cell MALDI-TOF mass spectrometry is a rapid tool for the identification and classification of microorganisms, now widely used even in clinical laboratories. However, its distinctive power is not sufficient for some closely-related species. The genus Cronobacter, formerly known as Enterobacter sakazakii, contains such species. In this work, a new method for the differentiation of five Cronobacter species is presented involving the tryptic digestion of cytoplasmatic proteins followed by MALDI mass spectrometry analysis. A database was developed for use in Bruker Biotyper software including 52 reference spectra and tested on a set of 45 samples with an overall accuracy of about 80%. The possibility of measurement automation and the short time and low cost requirements of this method compared to those of biochemical tests or PCR methods make it a supplementary option to intact cell MALDI, providing additional information about the differentiation of problematic species. •A new method for Cronobacter species differentiation was developed.•The method relies on proteolysis and MALDI-TOF mass spectrometry analysis.•A reference spectra database was developed for use in Bruker Biotyper software.•Five Cronobacter species have been differentiated.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2014.01.008