Published ahead of print on April 15, 2005, doi:10.1165/rcmb.2004-0352OC
© 2005 American Thoracic Society DOI: 10.1165/rcmb.2004-0352OC Identification of DNA Markers for a Transmissible Pseudomonas aeruginosa Cystic Fibrosis StrainCardiff School of Biosciences, Cardiff University, Cardiff, Wales; Manchester Adult Cystic Fibrosis Centre, Wythenshawe Hospital, Southmoor Road, Manchester; Pathogen Sequencing Unit, The Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge; Department of Medical Microbiology, University of Edinburgh Medical School, Teviot Place, Edinburgh, Scotland, United Kingdom; Division of Infectious and Immunological Diseases, Department of Pediatrics, University of British Columbia, Vancouver, British Columbia, Canada; Department of Pediatrics, University of Michigan, Ann Arbor, Michigan; and Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts Correspondence and requests for reprints should be addressed to Eshwar Mahenthiralingam, Ph.D, Senior Lecturer, Cardiff School of Biosciences, Main Building, Museum Avenue, Cardiff University, Cardiff, Wales CF10 3TL, UK. E-mail: MahenthiralingamE{at}cardiff.ac.uk A number of transmissible Pseudomonas 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 hybridization, 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 hybridization markers (MA15, MA18, MA21, MA22, and MA30) were found to be specific to strain MA and were located within a novel 13,318-bp genomic island, designated the MA island. The MA island encoded 18 genes and consisted of two bacteriophage-like regions; one region encoded the MA-specific subtractive hybridization markers, while the other bacteriophage-like region contained a Vibrio cholera-like toxin gene. Probes MA15, MA18, MA21, MA22, and MA30 were all found to be specific to strain MA when a collection of 141 P. aeruginosa strains was examined by hybridization with each DNA marker. In contrast, a previously isolated DNA marker for the Liverpool transmissible CF strain, PS21, was not found to be specific, detecting two additional strain types in the collection screened. Both the Manchester and Liverpool strain types were not encountered in CF populations outside the United Kingdom. The MA genomic island and Vibrio cholera–like toxin gene within it constitute novel genetic factors associated with a transmissible P. aeruginosa strain and their role in pathogenesis remains to be determined.
Key Words: cystic fibrosis • transmission • identification • genomic island This article has been cited by other articles:
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