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Journal of Clinical Microbiology, July 1999, p. 2201-2208, Vol. 37, No. 7
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Differentiation of Burkholderia Species by PCR-Restriction Fragment Length Polymorphism Analysis of the 16S rRNA Gene and Application to Cystic Fibrosis Isolates

Christine Segonds,1,* Thierry Heulin,2,dagger Nicole Marty,1 and Gerard Chabanon1

Laboratoire de Bactériologie-Virologie-Hygiène, CHU Rangueil, 31403 Toulouse Cedex 4,1 and Centre de Pédologie Biologique, UPR 6831 CNRS, 54501 Vandoeuvre-les-Nancy,2 France

Received 28 September 1998/Returned for modification 7 December 1998/Accepted 17 March 1999

Burkholderia cepacia, which is an important pathogen in cystic fibrosis (CF) owing to the potential severity of the infections and the high transmissibility of some clones, has been recently shown to be a complex of five genomic groups, i.e., genomovars I, II (B. multivorans), III, and IV and B. vietnamiensis. B. gladioli is also involved, though rarely, in CF. Since standard laboratory procedures fail to provide an accurate identification of these organisms, we assessed the ability of restriction fragment length polymorphism (RFLP) analysis of amplified 16S ribosomal DNA (rDNA), with the combination of the patterns obtained with six endonucleases, to differentiate Burkholderia species. This method was applied to 16 type and reference strains of the genus Burkholderia and to 51 presumed B. cepacia clinical isolates, each representative of one clone previously determined by PCR ribotyping. The 12 Burkholderia type strains tested were differentiated, including B. cepacia, B. multivorans, B. vietnamiensis, and B. gladioli, but neither the genomovar I and III reference strains nor the genomovar IV reference strain and B. pyrrociniaT were distinguishable. CF clinical isolates were mainly distributed in RFLP group 2 (which includes B. multivoransT) and RFLP group 1 (which includes B. cepacia genomovar I and III reference strains, as well as nosocomial clinical isolates). Two of the five highly transmissible clones in French CF centers belonged to RFLP group 2, and three belonged to RFLP group 1. The remaining isolates either clustered with other Burkholderia species (B. cepacia genomovar IV or B. pyrrocinia, B. vietnamiensis, and B. gladioli) or harbored unique combinations of patterns. Thus, if further validated by hybridization studies, PCR-RFLP of 16S rDNA could be an interesting identification tool and contribute to a better evaluation of the respective clinical risks associated with each Burkholderia species or genomovar in patients with CF.

* Corresponding author. Mailing address: Laboratoire de Bactériologie-Virologie-Hygiène, CHU Rangueil, 1 avenue Jean Poulhès, 31403 Toulouse Cedex 4, France. Phone: 33-5-61-32-21-55. Fax: 33-5-61-32-26-20. E-mail: segonds{at}cict.fr.

dagger Present address: Laboratoire d'Ecologie Microbienne de la Rhizosphère, DSV-DEVM, UMR CNRS-CEA, Commissariat à l'Energie Atomique, Centre de Cadarache, 13108 Saint Paul-lez-Durance Cedex, France.

Journal of Clinical Microbiology, July 1999, p. 2201-2208, Vol. 37, No. 7
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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