PCR-based detection of a cystic fibrosis epidemic strain of Pseudomonas Aeruginosa.
BACKGROUND
The Liverpool epidemic strain (LES) of Pseudomonas aeruginosa is widespread among patients with cystic fibrosis (CF) in specialist centers around Liverpool and elsewhere in the UK. This study evaluates a new diagnostic PCR assay based on a unique DNA sequence (PS21) of LES, for its identification of colonies directly from sputum.


METHODS
One hundred and fifty-eight sputum samples from 92 patients were cultured and P. aeruginosa isolates were typed by PS21 PCR and pulsed-field gel electrophoresis (PFGE). Subsequently, PS21 PCR was performed directly on sputum and the results were compared with culture, PFGE, and PS21 PCR typing.


RESULTS
Eighty patients were colonized with P. aeruginosa, 63 by LES (79%). There was 100% concordance between PS21 PCR on colonies and PFGE typing. The sensitivity and specificity of PS21 PCR directly on sputum was 98.2% and 93.6%, respectively.


CONCLUSIONS
This study shows that PS21 PCR can be used for simple and rapid screening of LES colonization in CF patients.
Panagea
Stavroula
S
Department of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Liverpool, UK. vivkypanagea@doctors.org.uk
Winstanley
Craig
C
Parsons
Yasmin N
YN
Walshaw
Martin J
MJ
Ledson
Martin J
MJ
Hart
C Anthony
CA
D001483:Base Sequence; D003550:Cystic Fibrosis / Q000382:microbiology*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D016521:Electrophoresis, Gel, Pulsed-Field; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D011552:Pseudomonas Infections / Q000453:epidemiology*; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D013183:Sputum / Q000382:microbiology
D017931:DNA Primers; D004269:DNA, Bacterial