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Journal of Clinical Microbiology, September 2004, p. 4121-4126, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.4121-4126.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Correlation of wbiI Genotype, Serotype, and Isolate Source within Species of the Burkholderia cepacia Complex

Arlene D. Vinion-Dubiel,1 Theodore Spilker,2 Charles R. Dean,1,{dagger} Henri Monteil,3 John J. LiPuma,2 and Joanna B. Goldberg1*

Department of Microbiology, University of Virginia Health Sciences Center, Charlottesville, Virginia,1 Department of Pediatrics and Communicable Diseases, University of Michigan Medical School, Ann Arbor, Michigan,2 Institute of Bacteriology, Strasbourg University Hospital, Louis Pasteur University, Strasbourg, France3

Received 6 February 2004/ Returned for modification 12 March 2004/ Accepted 20 April 2004

Gram-negative bacteria of the Burkholderia cepacia complex (Bcc) are opportunistic pathogens that can infect the lungs of cystic fibrosis (CF) patients and can be transmitted among these patients, causing epidemics in the CF community. Lipopolysaccharide (LPS) is an important virulence factor of many gram-negative bacteria, with the O antigen component of LPS being responsible for serotype specificity. The goal of this work was to develop a genetic method of determining the serotype of Bcc isolates based on the conserved gene wbiI. Homologues of wbiI are found in polysaccharide biosynthesis gene clusters in other bacteria. Primers to a conserved region of the Bcc wbiI gene were able to amplify by PCR a single product in 67 of 80 Bcc isolates tested. Sequencing and restriction enzyme digestion of this wbiI PCR product revealed sufficient DNA polymorphisms to distinguish and group various isolates. In five of nine instances, Bcc isolates of a single serotype had a single wbiI restriction fragment length polymorphism (RFLP) pattern, while isolates of the other four serotypes could have multiple wbiI RFLP types. Species determination of the Bcc isolates revealed no obvious correlation between wbiI RFLP type and species. There was also no apparent correlation between wbiI RFLP type and the ability of a single Bcc isolate to infect an individual with CF. However three of five Bcc outbreaks involved isolates with the same wbiI RFLP type, indicating that wbiI RFLP typing may be a useful tool to help track Bcc outbreaks.

* Corresponding author. Mailing address: Department of Microbiology, University of Virginia, P.O. Box 800734, Charlottesville, VA 22908-0734. Phone: (434) 243-2774. Fax: (434) 982-1071. E-mail: jbg2b{at}virginia.edu.

{dagger} Present address: Novartis Institute for Biomedical Research, Cambridge, Mass.

Journal of Clinical Microbiology, September 2004, p. 4121-4126, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.4121-4126.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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