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J. Clin. Microbiol. doi:10.1128/JCM.00702-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Multiple-locus variable-number tandem repeat analysis for the longitudinal survey of Pseudomonas aeruginosa in cystic fibrosis patients

Hôpital Armand Trousseau, Assistance Publique-Hôpitaux de Paris (APHP), Bactériologie and Pneumologie pédiatrie, Inserm UMR S- 719, 75012 Paris, France, Univ Paris-Sud, Institut de Génétique et Microbiologie, Orsay, F-91405, France; CNRS, Orsay, F-91405, France, Division de Microbiologie Analytique, Centre d'Etudes du Bouchet, Vert le Petit, France

^* To whom correspondence should be addressed. Email: christine.pourcel{at}u-psud.fr.

	Abstract

In order to identify the source of infection by Pseudomonas aeruginosa in patients with cystic fibrosis (CF), systematic genotyping of isolates is necessary. Multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) was used to survey P. aeruginosa infection in a French paediatric CF centre (Paris). Between January 2004 and December 2006, 108 patients, aged 2 to 21 years, regularly followed up at the centre, provided sputum for culture. P. aeruginosa was detected in 46 children, of which 19 had primary colonization. A total of 163 isolates were recovered. From a previously published method, MLVA was improved by the addition of new, informative and easily typable markers. Upon genotyping with 15 VNTRs, a total of 39 lineages composed of indistinguishable or closely related isolates, were observed. One of them corresponds to "clone C" widely distributed in Europe and another corresponds to the reference strain PA14. Six patients were colonized with 2 different strains and the remaining 40 patients were colonized with a single strain. Strains from seven lineages are shared by at least two and up to four patients in a total of 20 patients. The study demonstrates that MLVA is an efficient, easy and rapid molecular method of epidemiological surveillance for P. aeruginosa infection. The resulting data and strain genetic profiles can be queried on http://bacterial-genotyping.igmors.u-psud.fr.

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