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Infection and Immunity, February 2004, p. 1072-1083, Vol. 72, No. 2
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.2.1072-1083.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

New Aspects Regarding Evolution and Virulence of Listeria monocytogenes Revealed by Comparative Genomics and DNA Arrays

Michel Doumith,1 Christel Cazalet,2 Natalie Simoes,2 Lionel Frangeul,2 Christine Jacquet,1 Frank Kunst,2 Paul Martin,1 Pascale Cossart,3 Philippe Glaser,2 and Carmen Buchrieser2*

Laboratoire des Listeria, Centre National de Référence des Listeria, World Health Organization Collaborating Center for Foodborne Listeriosis,1 Laboratoire de Génomique des Microorganismes Pathogènes,2 Unité des Interactions Bactéries-Cellules, Institut Pasteur, 75724 Paris Cedex 15, France3

Received 12 September 2003/ Returned for modification 20 October 2003/ Accepted 22 October 2003

Listeria monocytogenes is a food-borne bacterial pathogen that causes a wide spectrum of diseases, such as meningitis, septicemia, abortion, and gastroenteritis, in humans and animals. Among the 13 L. monocytogenes serovars described, invasive disease is mostly associated with serovar 4b strains. To investigate the genetic diversity of L. monocytogenes strains with different virulence potentials, we partially sequenced an epidemic serovar 4b strain and compared it with the complete sequence of the nonepidemic L. monocytogenes EGDe serovar 1/2a strain. We identified an unexpected genetic divergence between the two strains, as about 8% of the sequences were serovar 4b specific. These sequences included seven genes coding for surface proteins, two of which belong to the internalin family, and three genes coding for transcriptional regulators, all of which might be important in different steps of the infectious process. Based on the sequence information, we then characterized the gene content of 113 Listeria strains by using a newly designed Listeria array containing the "flexible" part of the sequenced Listeria genomes. Hybridization results showed that all of the previously identified virulence factors of L. monocytogenes were present in the 93 L. monocytogenes strains tested. However, distinct patterns of the presence or absence of other genes were identified among the different L. monocytogenes serovars and Listeria species. These results allow new insights into the evolution of L. monocytogenes, suggesting that early divergence of the ancestral L. monocytogenes serovar 1/2c strains from the serovar 1/2b strains led to two major phylogenetic lineages, one of them including the serogroup 4 strains, which branched off the serovar 1/2b ancestral lineage, leading (mostly by gene loss) to the species Listeria innocua. The identification of 30 L. monocytogenes-specific and several serovar-specific marker genes, such as three L. monocytogenes serovar 4b-specific surface protein-coding genes, should prove powerful for the rapid tracing of listeriosis outbreaks, but it also represents a fundamental basis for the functional study of virulence differences between L. monocytogenes strains.

* Corresponding author. Mailing address: Laboratoire de Génom-ique des Microorganismes Pathogènes, Institut Pasteur, 28 rue du Dr. Roux, 75724 Paris Cedex 15, France. Phone: (33-1)-45-68-83-72. Fax: (33-1)-45-68-87-86. E-mail: cbuch{at}pasteur.fr.

Editor: J. T. Barbieri

Infection and Immunity, February 2004, p. 1072-1083, Vol. 72, No. 2
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.2.1072-1083.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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