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Int J Syst Evol Microbiol 53 (2003), 739-746; DOI  10.1099/ijs.0.02483-0
© 2003 International Union of Microbiological Societies

DNA–DNA hybridization study of Burkholderia species using genomic DNA macro-array analysis coupled to reverse genome probing

Vincent Ramisse1, Jacques Balandreau2, François Thibault3, Dominique Vidal3, Gilles Vergnaud1 and Philippe Normand2

1 Centre d'Etudes du Bouchet (CEB), BP3, 91710 Vert le Petit, France
2 Écologie Microbienne, UMR 5557 CNRS, Bât. 741, Université Claude Bernard Lyon I, 69622 Villeurbanne cedex, France
3 Centre de Recherches du Service de Santé des Armées, BP87, 38702 La Tronche cedex, France

Correspondence
Vincent Ramisse
vramisse{at}ceb.etca.fr

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.

Abbreviations: CF, cystic fibrosis; MF, microfluorimetry; NS1, nuclease S1; R, hybridization signal ratio; SP, spectrophotometry

Published online ahead of print on 19 September 2002 as DOI 10.1099/ijs.0.02483-0.

The EMBL/GenBank accession number for the 16S rRNA gene sequence of CEB 01056 is AJ491304.




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