Development of a diagnostic test for the Midlands 1 cystic fibrosis epidemic strain of Pseudomonas aeruginosa.
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.
Smart
Catherine H M
CHM
Division of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, UK.
Scott
Fiona W
FW
Laboratory of HealthCare Associated Infection, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK.
Wright
Elli A
EA
Division of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, UK.
Walshaw
Martin J
MJ
Regional Adult Cystic Fibrosis Unit, Cardiothoracic Centre, Liverpool L14 3PE, UK.
Hart
C Anthony
CA
Division of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, UK.
Pitt
Tyrone L
TL
Laboratory of HealthCare Associated Infection, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK.
Winstanley
Craig
C
Division of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, UK.
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