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Multilocus Sequence Typing Scheme That Provides Both Species and Strain Differentiation for the Burkholderia cepacia Complex

Department of Biological Sciences, Warwick University, Coventry CV4 7AL, United Kingdom,¹ Cardiff School of Biosciences, Cardiff University, Cardiff CF10 3TL, United Kingdom,² Department of Respiratory Medicine, Birmingham Heartlands Hospital, Birmingham, United Kingdom,³ Peter Medawar Building for Pathogen Research and Department of Zoology, University of Oxford, Oxford, United Kingdom,⁴ Department of Medical Microbiology, University of Edinburgh Medical School, Teviot Place, Edinburgh, Scotland, United Kingdom,⁵ Department of Pediatrics, University of British Columbia, Vancouver, British Columbia, Canada,⁶ Department of Pediatrics and Communicable Diseases, University of Michigan Medical School, 1150 W. Medical Center Drive, 8323 MRSB III, Box 0646, Ann Arbor, Michigan 48109-0646,⁷ Laboratorium voor Microbiologie, Universiteit Gent, Ledeganckstraat 35, B-9000 Ghent, Belgium⁸

Received 19 April 2005/ Returned for modification 25 May 2005/ Accepted 21 June 2005

A single multilocus sequence typing (MLST) scheme was developed for precise characterization of the opportunistic pathogens of Burkholderia cepacia complex (BCC), a group composed of at least nine closely related species. Seven conserved housekeeping genes were selected after a comparison of five Burkholderia species, and a collection of strains was subjected to nucleotide sequence analysis using a nested PCR amplification approach for each gene. MLST differentiated all nine current BCC species and identified 114 sequence types within a collection of 119 strains. No differentiation was found between strains recovered from environmental or clinical sources. The improved resolution in strain identification offered by MLST was able to identify previously characterized epidemic strain lineages and also demonstrated the presence of four novel potential species groups within the complex. There was also evidence for recombination having an important role in the recent evolution of individual BCC species. This highly transferable, validated, MLST scheme provides a new means to assist in species identification as well as unambiguous strain discrimination of the BCC by a single approach. It is also the first MLST scheme designed at the outset to incorporate multiple species and should facilitate global epidemiological investigations of the BCC.

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