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Journal of Clinical Microbiology, May 2003, p. 1819-1826, Vol. 41, No. 5
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.5.1819-1826.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of a Tandem Repeat Polymorphism in Legionella pneumophila and Its Use for Genotyping

C. Pourcel,1* Y. Vidgop,1 F. Ramisse,2 G. Vergnaud,1,2 and C. Tram3

Génome, Polymorphisme et Minisatellites (GPMS), Institut de Génétique et Microbiologie, Université Paris XI, 91405 Orsay Cedex,1 Centre d'Etude du Bouchet, 91710 Vert le Petit,2 Unité de Bactériologie Moléculaire et Médicale, Institut Pasteur, 75723 Paris Cedex 15, France3

Received 7 November 2002/ Returned for modification 21 January 2003/ Accepted 4 February 2003

We have analyzed the variability of minisatellite sequences (also called variable-number tandem repeats [VNTRs]) in the genome of Legionella pneumophila. Based upon the genome sequence of the Philadelphia-1 strain (serogroup 1), 25 minisatellites were selected and their polymorphisms were analyzed by PCR with the DNA of serogroup 1 to 14 reference strains. For 22 markers, a PCR product of the expected size was found with the DNA of the Philadelphia-1 strain. Most of these markers did not amplify the DNA of other Legionella species or other bacteria used as controls. A polymorphism was observed for seven markers among the L. pneumophila strains tested. To check whether these markers could be used to compare strains of L. pneumophila, we analyzed two groups of isolates from clinical and environmental samples which had been independently genotyped by other methods. The results showed that, for the isolates in these two sets of samples, VNTR typing is as informative as pulsed-field gel electrophoresis for comparison of strains. Sequencing of one minisatellite from 14 reference strains was performed. Comparison of the sequences allowed a classification and confirmed the existence of subspecies of L. pneumophila. We also tested the usefulness of one very polymorphic marker as a tool for the rapid screening of colonies grown from water samples. This allowed the rapid identification of the L. pneumophila colonies and gave a first hint as to the presence of several strains in a single sample.

* Corresponding author. Mailing address: GPMS, Institut de Génétique et Microbiologie, Bat. 400, Université Paris XI, 91405 Orsay cedex, France. Phone: (33) 1 69 15 30 01. Fax: (33) 1 69 15 66 78. E-mail: Christine.Pourcel{at}igmors.u-psud.fr.

Journal of Clinical Microbiology, May 2003, p. 1819-1826, Vol. 41, No. 5
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.5.1819-1826.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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