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Enzymatic protein digests do not assist in E. coli discrimination at the strain level using mass spectrometry

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Author(s): Ricardo J. Carreira | J. D. Nunes-Miranda | Alexandre Gonçalves | Gilberto Igrejas | Patrícia Poeta | Silvana Gómez-Meire | Miguel Reboiro-Jato | Florentino Fdez-Riverola | Carlos Lodeiro | José-Luis Capelo-Martínez

Journal: Journal of Integrated OMICS
ISSN 2182-0287

Volume: 3;
Issue: 1;
Start page: 44;
Date: 2013;
Original page

Keywords: Bacterial | E. Coli | Fingerprinting | MALDI | Classification

ABSTRACT
Different procedures for matrix assisted laser desorption ionization time of flight mass spectrometry-based E. coli classification at the strain level using the enzymatic digestion of proteins from the cell lysate have been studied. The effects of ultrasonic energy as well as the effects of protein reduction and protein alkylation in the sample treatment and in the subsequent classification were assessed. The final optimal method for classification was then compared with an intact cell-based approach in a different set of samples. Our results show that E. coli classification at the strain level is possible as 12 different strains were correctly classified using intact cell analysis. Overall, the confidence level in classification was higher when the analysis was performed with the intact cell approach.
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