Identification of variable-number tandem-repeat (VNTR) sequences in Legionella pneumophila and development of an optimized multiple-locus VNTR analysis typing scheme.
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/.
Pourcel
Christine
C
Institut de Génétique et Microbiologie, Bât 400, Université Paris-Sud, 91405 Orsay cedex, France. christine.pourcel@igmors.u-psud.fr
Visca
Paolo
P
Afshar
Baharak
B
D'Arezzo
Silvia
S
Vergnaud
Gilles
G
Fry
Norman K
NK
D015373:Bacterial Typing Techniques / Q000379:methods*; D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D005838:Genotype; D006801:Humans; D016952:Legionella pneumophila / Q000145:classification* / Q000235:genetics*; D007877:Legionnaires' Disease / Q000453:epidemiology / Q000382:microbiology; D018598:Minisatellite Repeats; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004269:DNA, Bacterial