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Applied and Environmental Microbiology, December 2008, p. 7767-7778, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.01402-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Rapid Classification and Identification of Salmonellae at the Species and Subspecies Levels by Whole-Cell Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry{triangledown} ,{dagger}

Ralf Dieckmann,1 Reiner Helmuth,1 Marcel Erhard,2 and Burkhard Malorny1*

Federal Institute for Risk Assessment, National Salmonella Reference Laboratory, Berlin, Germany,1 Anagnostec GmbH, Am Mühlenberg 11, D-14476 Potsdam-Golm, Germany2

Received 23 June 2008/ Accepted 16 October 2008

Variations in the mass spectral profiles of multiple housekeeping proteins of 126 strains representing Salmonella enterica subsp. enterica (subspecies I), S. enterica subsp. salamae (subspecies II), S. enterica subsp. arizonae (subspecies IIIa), S. enterica subsp. diarizonae (subspecies IIIb), S. enterica subsp. houtenae (subspecies IV), and S. enterica subsp. indica (subspecies VI), and Salmonella bongori were analyzed to obtain a phylogenetic classification of salmonellae based on whole-cell matrix-assisted laser desorption ionization-time of flight mass spectrometric bacterial typing. Sinapinic acid produced highly informative spectra containing a large number of biomarkers and covering a wide molecular mass range (2,000 to 40,000 Da). Genus-, species-, and subspecies-identifying biomarker ions were assigned on the basis of available genome sequence data for Salmonella, and more than 200 biomarker peaks, which corresponded mainly to abundant and highly basic ribosomal or nucleic acid binding proteins, were selected. A detailed comparative analysis of the biomarker profiles of Salmonella strains revealed sequence variations corresponding to single or multiple amino acid changes in multiple housekeeping proteins. The resulting mass spectrometry-based bacterial classification was very comparable to the results of DNA sequence-based methods. A rapid protocol that allowed identification of Salmonella subspecies in minutes was established.

* Corresponding author. Mailing address: Federal Institute for Risk Assessment, National Salmonella Reference Laboratory, Diedersdorfer Weg 1, D-12277 Berlin, Germany. Phone: 49-30-8412-2237. Fax: 49-30-8412-2953. E-mail: burkhard.malorny{at}bfr.bund.de

{triangledown} Published ahead of print on 24 October 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, December 2008, p. 7767-7778, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.01402-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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