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Published ahead of print on April 15, 2005, doi:10.1165/rcmb.2004-0352OC

Am. J. Respir. Cell Mol. Biol., Volume 33, Number 1, July 2005, 56-64

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Submitted on November 10, 2004
Revised on April 15, 2005

Identification of DNA markers for a transmissible Pseudomonas aeruginosa cystic fibrosis strain

Deborah A Lewis1, Andrew Jones2, Julian Parkhill3, David P Speert4, John R.W. Govan5, John J LiPuma6, Stephen Lory7, A. Kevin Webb2, and Eshwar Mahenthiralingam8*

1 Cardiff School of Biosciences, University of Cardiff, Cardiff, Wales, United Kingdom; Faculty of Medical and Veterinary Sciences, University of Bristol, Bristol, Avon, United Kingdom, 2 Manchester Adult Cystic Fibrosis Centre, Wythenshawe Hospital, Manchester, Lancashire, United Kingdom, 3 Pathogen Sequencing Unit, The Sanger Institute, Cambridge, Cambridgeshire, United Kingdom, 4 Department of Pediatrics, Division of Infectious and Immunological Diseases, University of British Columbia, Vancouver, British Columbia, Canada, 5 Department of Medical Microbiology, University of Edinburgh Medical School, Edinburgh, Scotland, United Kingdom, 6 Department of Pediatrics, University of Michigan, Ann Arbor, Michigan, USA, 7 Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts, USA, 8 Cardiff School of Biosciences, University of Cardiff, Cardiff, Wales, United Kingdom

* To whom correspondence should be addressed. E-mail: MahenthiralingamE{at}cardiff.ac.uk.

A number of transmissible P. aeruginosa strains have been identified which potentially constitute an emerging threat to patients with cystic fibrosis (CF). We sought to identify DNA markers that were specific to a transmissible P. aeruginosa CF clone and evaluate these probes on a large collection of genotypically distinct P. aeruginosa strains. Using subtractive DNA hybridisation, in combination with analysis using the P. aeruginosa PAO1 genome chip, DNA markers specific for or absent from the Manchester transmissible CF strain (MA) were identified. Five subtractive DNA hybridisation markers were located within a novel 13,318 bp genomic island, designated the MA island. The MA island encoded 18 genes, including a Vibrio cholera-like toxin gene and consisted of two bacteriophage-like regions. All five MA island markers were found to be specific to strain MA when a collection of 141 P. aeruginosa strains was examined. In contrast, a DNA marker for the Liverpool transmissible CF strain, PS21, was not found to be specific. Both the Manchester and Liverpool strain types were not encountered in CF populations outside the United Kingdom. The MA genomic island and V. cholera-like toxin gene within it constitute novel genetic factors associated with a transmissible P. aeruginosa strain.




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