Molecular Diagnosis

, Volume 7, Issue 3, pp 195–200 | Cite as

PCR-Based Detection of a Cystic Fibrosis Epidemic Strain of Pseudomonas Aeruginosa

  • Stavroula PanageaEmail author
  • Craig Winstanley
  • Yasmin N. Parsons
  • Martin J. Walshaw
  • Martin J. Ledson
  • C.Anthony Hart
Original Research Article


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.


Cystic Fibrosis Cystic Fibrosis Patient Sputum Sample Epidemic Strain PFGE Typing 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank J.E. Corkill for assistance with pulsed-field gel electrophoresis typing. Dr Panagea acknowledges funding from the Association of Medical Microbiologists, UK. Drs Winstanley and Hart acknowledge funding from the United Kingdom Cystic Fibrosis Trust and the Welcome Trust. This work was also supported by an R&D Support fund from Royal Liverpool Broadgreen University Hospitals Trust.

The authors have no conflicts of interest directly relevant to the content of this study.


  1. 1.
    Hart CA, Winstanley C. Persistent and aggressive bacteria in the lungs of cystic fibrosis children. Br Med Bull 2002; 61: 81–96PubMedCrossRefGoogle Scholar
  2. 2.
    Govan JRW. Infection control in cystic fibrosis: methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa and the Burkholderia cepacia complex. J R Soc Med 2000; 93 Suppl. 38: 40–5PubMedGoogle Scholar
  3. 3.
    Armstrong DS, Nixon GM, Carzino R, et al. Detection of a widespread clone of Pseudomonas aeruginosa in a pediatric cystic fibrosis clinic. Am J Respir Crit Care Med 2002; 166: 983–7PubMedCrossRefGoogle Scholar
  4. 4.
    Jones MA, Govan JRW, Doherty CJ, et al. Spread of a multiresistant strain of Pseudomonas aeruginosa in an adult cystic fibrosis clinic. Lancet 2001; 358: 557–8PubMedCrossRefGoogle Scholar
  5. 5.
    Cheng K, Smyth RL, Govan JRW, et al. Spread of a β-lactam-resistant Pseudomonas aeruginosa in a cystic fibrosis clinic. Lancet 1996; 348: 639–42PubMedCrossRefGoogle Scholar
  6. 6.
    McCallum SJ, Corkill JE, Gallagher M, et al. Superinfection with a transmissible strain of Pseudomonas aeruginosa in adults with cystic fibrosis chronically colonised by P. aeruginosa. Lancet 2001; 358: 558–60PubMedCrossRefGoogle Scholar
  7. 7.
    McCallum SJ, Gallagher MJ, Corkill JE, et al. Spread of an epidemic Pseudomonas. aeruginosa strain from a patient with cystic fibrosis (CF) to non-CF relatives. Thorax 2002; 57: 559–60Google Scholar
  8. 8.
    Scott F, Pitt TL. Identification and characterisation of transmissible Pseudomonas aeruginosa strains in UK cystic fibrosis centres [abstract 146]. J Cystic Fibrosis 2003; 2 Suppl. 1: 39Google Scholar
  9. 9.
    Cystic Fibrosis Trust Control of Infection Group. Pseudomonas aeruginosa infection in people with cystic fibrosis: suggestions for prevention and infection control. In: Report of the CF Trust’s Control of infection Group. London: CF Trust (UK), Cystic Fibrosis Trust, 2001 MayGoogle Scholar
  10. 10.
    The International Pseudomonas aeruginosa Typing Study Group. A multicenter comparison of methods for typing strains of Pseudomonas aeruginosa predominantly from patients with cystic fibrosis. J Infect Dis 1994 Jan; 169: 134–42CrossRefGoogle Scholar
  11. 11.
    Parsons YN, Panagea S, Smart CHM, et al. Use of subtractive hybridisation to identify a diagnostic probe for a cystic fibrosis epidemic strain of Pseudomonas aeruginosa. J Clin Microbiol 2002 Dec; 40: 4607–11PubMedCrossRefGoogle Scholar
  12. 12.
    Antimicrobial susceptibility testing: British Society for Antimicrobial Chemotherapy Working Party Report. J Antimicrob Chemother 2001; 48 Suppl 1: 43–57CrossRefGoogle Scholar
  13. 13.
    Ledson MJ, Gallagher MJ, Corkill JE, et al. Cross infection between cystic fibrosis patients colonised with Burkholderia cepacia. Thorax 1998; 53: 432–6PubMedCrossRefGoogle Scholar
  14. 14.
    Tenover FC, Arbeit RD, Goering RV, et al. Interpreting chromosomal DNA restriction patterns produced by Pulsed-Field Gel electrophoresis: criteria for bacterial typing. J Clin Microbiol 1996 Sep; 33: 2233–9Google Scholar
  15. 15.
    Khan AA, Cerniglia CE. Detection of Pseudomonas aeruginosa from clinical and environmental samples by amplification of the exotoxin A gene using PCR. Appl Environ Microbiol 1994 Oct; 60: 3739–45PubMedGoogle Scholar
  16. 16.
    Campbell III PW, Phillips III JA, Heidecker GJ, et al. Detection of Pseudomonas (Burkholderia) cepacia using PCR. Pediatr Pulmonol 1995; 20: 44–9PubMedCrossRefGoogle Scholar
  17. 17.
    Grothues D, Kopman U, von der Hardt H, et al. Genome fingerprinting of Pseudomonas aeruginosa indicates colonisation of cystic fibrosis siblings with closely related strains. J Clin Microbiol 1988 Oct; 26: 1973–7PubMedGoogle Scholar
  18. 18.
    Grundmann H, Scheider C, Hartung D, et al. Discriminatory power of three DNA-based typing techniques for Pseudomonas aeruginosa. J Clin Microbiol 1995 Mar; 33: 528–34PubMedGoogle Scholar
  19. 19.
    Romling U, Fielder B, Bobhammer J, et al. Epidemiology of chronic Pseudomonas aeruginosa infections in cystic fibrosis. J Infect Dis 1994 Dec; 170: 1616–21PubMedCrossRefGoogle Scholar
  20. 20.
    McIntosh I, Govan JRW, Brock DJH. Detection of Pseudomonas aeruginosa in sputum from cystic fibrosis patients by the polymerase chain reaction. Mol Cell Probes 1992; 6: 299–304PubMedCrossRefGoogle Scholar
  21. 21.
    Kaprati F, Jonasson J. Polymerase chain reaction for the detection of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Burkholderia cepacia in sputum of patients with cystic fibrosis. Mol Cell Probes 1996; 10: 397–403CrossRefGoogle Scholar
  22. 22.
    McDowell A, Mahenthiralingam E, Moore JE, et al. PCR-based detection and identification of Burkholderia cepacia complex pathogens in sputum from cystic fibrosis patients. J Clin Microbiol 2001 Dec; 39: 4247–55PubMedCrossRefGoogle Scholar
  23. 23.
    Geddes DM. Of isolates and isolation: Pseudomonas aeruginosa in adults with cystic fibrosis. Lancet 2001; 358: 522–3PubMedCrossRefGoogle Scholar
  24. 24.
    Ramsey BW. To cohort or not to cohort: how transmissible is Pseudomonas aeruginosa? Am J Respir Crit Care Med 2002; 166: 906–7PubMedCrossRefGoogle Scholar
  25. 25.
    Pitt TL. Cross infection of cystic fibrosis patients with Pseudomonas aeruginosa [editorial]. Thorax 2002; 57: 921PubMedCrossRefGoogle Scholar

Copyright information

© Adis Data Information BV 2003

Authors and Affiliations

  • Stavroula Panagea
    • 1
    Email author
  • Craig Winstanley
    • 1
  • Yasmin N. Parsons
    • 1
  • Martin J. Walshaw
    • 2
  • Martin J. Ledson
    • 2
  • C.Anthony Hart
    • 1
  1. 1.Department of Medical Microbiology and Genitourinary MedicineUniversity of LiverpoolLiverpoolUK
  2. 2.Regional Adult Cystic Fibrosis UnitCardiothoracic CentreLiverpoolUK

Personalised recommendations