title: Development of a diagnostic test for the Midlands 1 cystic fibrosis epidemic strain of Pseudomonas aeruginosa.
abstract: In a previous study of isolates from cystic fibrosis (CF) patients in England and Wales, the Midlands 1 strain of Pseudomonas aeruginosa was identified as the second most common clone, representing 10% of isolates and found in nearly one-third of all CF centres [Scott, F. W. & Pitt, T. L. (2004). J Med Microbiol 53, 609-615]. Using suppression subtractive hybridization, 54 sequences were identified as present in a Midlands 1 strain but were absent from strain PAO1. The distribution of 14 of these sequences amongst representatives of Midlands 1, other CF epidemic strains and unrelated P. aeruginosa CF isolates was determined using PCR assays. Using these data, a PCR-based test was developed that was specific for the Midlands 1 clone, which was confirmed using dot-blot hybridization. By applying the test to CF isolates from a CF centre in Liverpool, a Midlands 1 clone was identified. The identity was confirmed using typing by PFGE. The PCR test should facilitate a greater understanding of the distribution of the Midlands 1 strain in the UK and elsewhere.
authors.affiliation: Division of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, UK.
authors.forename: Catherine H M
authors.initials: CHM
authors.lastname: Smart
authors.affiliation: Laboratory of HealthCare Associated Infection, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK.
authors.forename: Fiona W
authors.initials: FW
authors.lastname: Scott
authors.affiliation: Division of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, UK.
authors.forename: Elli A
authors.initials: EA
authors.lastname: Wright
authors.affiliation: Regional Adult Cystic Fibrosis Unit, Cardiothoracic Centre, Liverpool L14 3PE, UK.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Walshaw
authors.affiliation: Division of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, UK.
authors.forename: C Anthony
authors.initials: CA
authors.lastname: Hart
authors.affiliation: Laboratory of HealthCare Associated Infection, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK.
authors.forename: Tyrone L
authors.initials: TL
authors.lastname: Pitt
authors.affiliation: Division of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, UK.
authors.forename: Craig
authors.initials: C
authors.lastname: Winstanley
mesh_terms: D003550:Cystic Fibrosis / Q000150:complications*; D016680:Genome, Bacterial; D006801:Humans; D017403:In Situ Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D011552:Pseudomonas Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000209:etiology; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D017421:Sequence Analysis; D013045:Species Specificity; D006113:United Kingdom / Q000453:epidemiology