title: Development of a multiple-locus variable-number tandem-repeat typing scheme for genetic fingerprinting of Burkholderia cenocepacia and application to nationwide epidemiological analysis.
abstract: Organisms of the Burkholderia cepacia complex are especially important pathogens in cystic fibrosis (CF), with a propensity for patient-to-patient spread and long-term respiratory colonization. B. cenocepacia and Burkholderia multivorans account for the majority of infections in CF, and major epidemic clones have been recognized throughout the world. The aim of the present study was to develop and evaluate a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) scheme for B. cenocepacia. Potential VNTR loci were identified upon analysis of the annotated genome sequences of B. cenocepacia strains AU1054, J2315, and MCO-3, and 10 of them were selected on the basis of polymorphisms and size. A collection of 100 B. cenocepacia strains, including epidemiologically related and unrelated strains, as well as representatives of the major epidemic lineages, was used to evaluate typeability, epidemiological concordance, and the discriminatory power of MLVA-10 compared with those of pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Longitudinal stability was assessed by testing 39 successive isolates from 14 patients. Typeability ranged from 0.91 to 1, except for that of one marker, which was not amplified in 53% of the B. cenocepacia IIIA strains. The MLVA types were shown to be stable in chronically colonized patients and within outbreak-related strains, with excellent epidemiological concordance. Epidemic and/or globally distributed lineages (epidemic Edinburgh-Toronto electrophoretic type 12 [ET-12], sequence type 32 [ST-32], ST-122, ST-234, and ST-241) were successfully identified. Conversely, the discriminatory power of MLVA was lower than that of PFGE or MLST, although PFGE variations within the epidemic lineages sometimes masked their genetic relatedness. In conclusion, MLVA represents a promising cost-effective first-line tool in B. cenocepacia surveillance.
authors.affiliation: Laboratoire de Bactériologie-Hygiène, Hôpital Purpan, CHU Toulouse, Toulouse, France U.1043 Inserm, UMR1043, Toulouse, France segonds.c@chu-toulouse.fr.
authors.forename: Christine
authors.initials: C
authors.lastname: Segonds
authors.affiliation: Service de Bactériologie et Hygiène Hospitalière, CHU Besançon, Besançon, France.
authors.forename: Michelle
authors.initials: M
authors.lastname: Thouverez
authors.affiliation: Laboratoire de Bactériologie-Hygiène, Hôpital Purpan, CHU Toulouse, Toulouse, France.
authors.forename: Antoine
authors.initials: A
authors.lastname: Barthe
authors.affiliation: U.1043 Inserm, UMR1043, Toulouse, France Université Toulouse III-Paul Sabatier, Toulouse, France.
authors.forename: Nadège
authors.initials: N
authors.lastname: Bossuet-Greif
authors.affiliation: U.1043 Inserm, UMR1043, Toulouse, France Université Toulouse III-Paul Sabatier, Toulouse, France.
authors.forename: Lenka
authors.initials: L
authors.lastname: Tisseyre
authors.affiliation: Service de Bactériologie et Hygiène Hospitalière, CHU Besançon, Besançon, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Plésiat
authors.affiliation: Université Paris-Sud, Institut de Génétique et Microbiologie, UMR 8621, Orsay, France CNRS, Orsay, France Université Paris-Saclay, ENSTA ParisTech, Palaiseau, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Vergnaud
authors.affiliation: Université Toulouse III-Paul Sabatier, Toulouse, France.
authors.forename: Gérard
authors.initials: G
authors.lastname: Chabanon
authors.affiliation: Université Paris-Sud, Institut de Génétique et Microbiologie, UMR 8621, Orsay, France CNRS, Orsay, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Pourcel
mesh_terms: D019121:Burkholderia Infections / Q000453:epidemiology / Q000382:microbiology*; D016956:Burkholderia cepacia / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D016000:Cluster Analysis; D019295:Computational Biology; D003550:Cystic Fibrosis / Q000150:complications; D016172:DNA Fingerprinting / Q000379:methods*; D014644:Genetic Variation; D016680:Genome, Bacterial; D005838:Genotype; D006801:Humans; D018598:Minisatellite Repeats; D017720:Molecular Epidemiology / Q000379:methods; D058889:Molecular Typing / Q000379:methods*