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Journal of Clinical Microbiology, April 2007, p. 1190-1199, Vol. 45, No. 4
0095-1137/07/$08.00+0     doi:10.1128/JCM.02078-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of Variable-Number Tandem-Repeat (VNTR) Sequences in Legionella pneumophila and Development of an Optimized Multiple-Locus VNTR Analysis Typing Scheme{triangledown}

Christine Pourcel,1* Paolo Visca,2,3 Baharak Afshar,4 Silvia D'Arezzo,2 Gilles Vergnaud,1,5 and Norman K. Fry4

Univ Paris-Sud, Institut de Génétique et Microbiologie, CNRS, Orsay F-91405, France,1 Istituto Nazionale per le Malattie Infettive Lazzaro Spallanzani I.R.C.C.S,2 Dipartimento di Biologia, Università Roma Tre, Rome, Italy,3 Health Protection Agency, Respiratory and Systemic Infection Laboratory, Centre for Infections, London, United Kingdom,4 Division de Microbiologie Analytique, Centre d'Etudes du Bouchet, Vert le Petit, France5

Received 9 October 2006/ Returned for modification 29 November 2006/ Accepted 17 January 2007

The utility of a genotypic typing assay for Legionella pneumophila was investigated. A multiple-locus variable number of tandem repeats (VNTR) analysis (MLVA) scheme using PCR and agarose gel electrophoresis is proposed based on eight minisatellite markers. Panels of well-characterized strains were examined in a multicenter analysis to validate the assay and to compare its performance to that of other genotyping assays. Excellent typeability, reproducibility, stability, and epidemiological concordance were observed. The MLVA type or profile is composed of a string of allele numbers, corresponding to the number of repeats at each VNTR locus, separated by commas, in a predetermined order. A database containing information from 99 L. pneumophila serogroup 1 strains and four strains of other serogroups and their MLVA profiles, which can be queried online, is available from http://bacterial-genotyping.igmors.u-psud.fr/.

* Corresponding author. Mailing address: Institut de Génétique et Microbiologie, Bât 400, Université Paris-Sud, 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

{triangledown} Published ahead of print on 24 January 2007.

Journal of Clinical Microbiology, April 2007, p. 1190-1199, Vol. 45, No. 4
0095-1137/07/$08.00+0     doi:10.1128/JCM.02078-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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