Distinguishing Species of the Burkholderia cepacia Complex and Burkholderia gladioli by Automated Ribotyping

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Journal of Clinical Microbiology, May 2000, p. 1876-1884, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Distinguishing Species of the Burkholderia cepacia Complex and Burkholderia gladioli by Automated Ribotyping

Sylvain Brisse,¹ Cees M. Verduin,² Dana Milatovic,¹ Ad Fluit,¹ Jan Verhoef,¹ Severine Laevens,³ Peter Vandamme,³ Burkhard Tümmler,⁴ Henri A. Verbrugh,² and Alex van Belkum²^,^*

Eijkman-Winkler Institute for Microbiology, Infectious Diseases and Inflammation, University Medical Centre Utrecht, 3584 CX Utrecht,¹ and Department of Medical Microbiology and Infectious Diseases, Erasmus University Medical Center Rotterdam, 3015 GD Rotterdam,² The Netherlands; Laboratory for Microbiology, University of Gent, B-9000 Gent, Belgium³; and Klinische Forscher Gruppe, Medizinische Hochschule Hannover, 30623 Hannover, Germany⁴

Received 22 November 1999/Returned for modification 2 February 2000/Accepted 11 February 2000

Several species belonging to the genus Burkholderia are clinically relevant, opportunistic pathogens that inhabit major environmentalreservoirs. Consequently, the availability of means for adequateidentification and epidemiological characterization of individualenvironmental or clinical isolates is mandatory. In the presentcommunication we describe the use of the Riboprinter microbialcharacterization system (Qualicon, Warwick, United Kingdom) forautomated ribotyping of 104 strains of Burkholderia species fromdiverse sources, including several publicly accessible collections.The main outcome of this analysis was that all strains were typeableand that strains of Burkholderia gladioli and of each speciesof the B. cepacia complex, including B. multivorans, B. stabilis,and B. vietnamiensis, were effectively discriminated. Furthermore,different ribotypes were discerned within each species. Ribotypingresults were in general agreement with strain classification basedon restriction fragment analysis of 16S ribosomal amplicons, butthe resolution of ribotyping was much higher. This enabled automatedmolecular typing below the species level. Cluster analysis ofthe patterns obtained by ribotyping (riboprints) showed that withinB. gladioli, B. multivorans, and B. cepacia genomovar VI, thedifferent riboprints identified always clustered together. Riboprintsof B. cepacia genomovars I and III, B. stabilis, and B. vietnamiensisdid not show distinct clustering but rather exhibited the formationof loose assemblages within which several smaller, genomovar-specificclusters were delineated. Therefore, ribotyping proved usefulfor genomovar identification. Analysis of serial isolates fromindividual patients demonstrated that infection with a singleribotype had occurred, despite minor genetic differences thatwere detected by pulsed-field gel electrophoresis of DNA macrorestrictionfragments. The automated approach allows very rapid and reliableidentification and epidemiological characterization of strainsand generates an easily manageable database suited for expansionwith information on additional bacterialisolates.

^* Corresponding author. Mailing address: Department of Medical Microbiology and Infectious Diseases, Erasmus University MedicalCenter Rotterdam (EMCR), Dr. Molewaterplein 40, 3015 GD Rotterdam,The Netherlands. Phone: 31-10-4635813. Fax: 31-10-4633875. E-mail:vanbelkum{at}bacl.azr.nl.

Journal of Clinical Microbiology, May 2000, p. 1876-1884, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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