DNA-DNA hybridization study of Burkholderia species using genomic DNA macro-array analysis coupled to reverse genome probing.
The present study was aimed at simplifying procedures to delineate species and identify isolates based on DNA-DNA reassociation. DNA macro-arrays harbouring genomic DNA of reference strains of several Burkholderia species were produced. Labelled genomic DNA, hybridized to such an array, allowed multiple relative pairwise comparisons. Based on the relative DNA-DNA relatedness values, a complete data matrix was constructed and the ability of the method to discriminate strains belonging to different species was assessed. This simple approach led successfully to the discrimination of Burkholderia mallei from Burkholderia pseudomallei, but also discriminated Burkholderia cepacia genomovars I and III, Burkholderia multivorans, Burkholderia pyrrocinia, Burkholderia stabilis and Burkholderia vietnamiensis. Present data showed a sufficient degree of congruence with previous DNA-DNA reassociation techniques. As part of a polyphasic taxonomic scheme, this straightforward approach is proposed to improve species definition, especially for application in the rapid screening necessary for large numbers of clinical or environmental isolates.
Ramisse
Vincent
V
Centre d'Etudes du Bouchet (CEB), BP3, 91710 Vert le Petit, France.
Balandreau
Jacques
J
Écologie Microbienne, UMR 5557 CNRS, Bât. 741, Université Claude Bernard Lyon I, 69622 Villeurbanne cedex, France.
Thibault
François
F
Centre de Recherches du Service de Santé des Armées, BP87, 38702 La Tronche cedex, France.
Vidal
Dominique
D
Centre de Recherches du Service de Santé des Armées, BP87, 38702 La Tronche cedex, France.
Vergnaud
Gilles
G
Centre d'Etudes du Bouchet (CEB), BP3, 91710 Vert le Petit, France.
Normand
Philippe
P
Écologie Microbienne, UMR 5557 CNRS, Bât. 741, Université Claude Bernard Lyon I, 69622 Villeurbanne cedex, France.
D015373:Bacterial Typing Techniques; D019117:Burkholderia / Q000145:classification* / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal; D020459:Genes, rRNA; D016680:Genome, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D012336:RNA, Ribosomal, 16S; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S