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Applied and Environmental Microbiology, September 2008, p. 5402-5407, Vol. 74, No. 17
0099-2240/08/$08.00+0     doi:10.1128/AEM.02689-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Rapid Identification and Typing of Listeria Species by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry{triangledown} ,{dagger}

Sukhadeo B. Barbuddhe,1 Thomas Maier,2 Gerold Schwarz,2 Markus Kostrzewa,2 Herbert Hof,3 Eugen Domann,1 Trinad Chakraborty,1 and Torsten Hain1*

Institute of Medical Microbiology, Justus Liebig University, Frankfurter Strasse 107, D-35392 Giessen, Germany,1 Bruker Daltonik GmbH, Permoserstrasse 15, D-04318 Leipzig, Germany,2 University Hospital Mannheim, Institute of Medical Microbiology, Thedor-Kutzer-Ufer 1-3, D-68167 Mannheim, Germany3

Received 28 November 2007/ Accepted 30 June 2008

Listeria monocytogenes is a food-borne pathogen that is the causative agent of human listeriosis, an opportunistic infection that primarily infects pregnant women and immunologically compromised individuals. Rapid, accurate discrimination between Listeria strains is essential for appropriate therapeutic management and timely intervention for infection control. A rapid method involving matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) that shows promise for identification of Listeria species and typing and even allows for differentiation at the level of clonal lineages among pathogenic strains of L. monocytogenes is presented. A total of 146 strains of different Listeria species and serotypes as well as clinical isolates were analyzed. The method was compared with the pulsed-field gel electrophoresis analysis of 48 Listeria strains comprising L. monocytogenes strains isolated from food-borne epidemics and sporadic cases, isolates representing different serotypes, and a number of Listeria strains whose genomes have been completely sequenced. Following a short inactivation/extraction procedure, cell material from a bacterial colony was deposited on a sample target, dried, overlaid with a matrix necessary for the MALDI process, and analyzed by MALDI-TOF MS. This technique examines the chemistry of major proteins, yielding profile spectra consisting of a series of peaks, a characteristic "fingerprint" mainly derived from ribosomal proteins. Specimens can be prepared in a few minutes from plate or liquid cultures, and a spectrum can be obtained within 1 minute. Mass spectra derived from Listeria isolates showed characteristic peaks, conserved at both the species and lineage levels. MALDI-TOF MS fingerprinting may have potential for Listeria identification and subtyping and may improve infection control measures.

* Corresponding author. Mailing address: Institute of Medical Microbiology, Justus Liebig University, Frankfurter Strasse 107, D-35392 Giessen, Germany. Phone: 49-641 99 46400. Fax: 49-641 99 46409. E-mail: Torsten.Hain{at}mikrobio.med.uni-giessen.de

{triangledown} Published ahead of print on 7 July 2008.

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

Applied and Environmental Microbiology, September 2008, p. 5402-5407, Vol. 74, No. 17
0099-2240/08/$08.00+0     doi:10.1128/AEM.02689-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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