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Rapid genotyping of Pseudomonas aeruginosa isolates harboured by adult and paediatric patients with cystic fibrosis using repetitive-element-based PCR assays

¹Clinical Virology and Molecular Microbiology Research Unit, Sir Albert Sakzewksi Virus Research Centre, Royal Children's Hospital and Clinical Medical Virology Centre, University of Queensland, Herston, Queensland, Australia 4029 ^2,6Department of Paediatrics and Child Health² and Department of Medicine⁶, University of Queensland, Brisbane, Queensland, Australia ³Adult Cystic Fibrosis Unit, The Prince Charles Hospital, Brisbane, Queensland, Australia ⁴Department of Respiratory Medicine, Royal Children's Hospital, Brisbane, Queensland, Australia ⁵Department of Microbiology, Queensland Health Pathology Service, The Prince Charles Hospital Campus, Brisbane, Queensland, Australia

Correspondence Michael D. Nissen theniss{at}uq.edu.au

Received January 22, 2004
Accepted July 7, 2004

In this study, the suitability of two repetitive-element-based PCR (rep-PCR) assays, enterobacterial repetitive intergenic consensus (ERIC)-PCR and BOX-PCR, to rapidly characterize Pseudomonas aeruginosa strains isolated from patients with cystic fibrosis (CF) was examined. ERIC-PCR utilizes paired sequence-specific primers and BOX-PCR a single primer that target highly conserved repetitive elements in the P. aeruginosa genome. Using these rep-PCR assays, 163 P. aeruginosa isolates cultured from sputa collected from 50 patients attending an adult CF clinic and 50 children attending a paediatric CF clinic were typed. The results of the rep-PCR assays were compared to the results of PFGE. All three assays revealed the presence of six major clonal groups shared by multiple patients attending either of the CF clinics, with the dominant clonal group infecting 38 % of all patients. This dominant clonal group was not related to the dominant clonal group detected in Sydney or Melbourne (pulsotype 1), nor was it related to the dominant groups detected in the UK. In all, PFGE and rep-PCR identified 58 distinct clonal groups, with only three of these shared between the two clinics. The results of this study showed that both ERIC-PCR and BOX-PCR are rapid, highly discriminatory and reproducible assays that proved to be powerful surveillance screening tools for the typing of clinical P. aeruginosa isolates recovered from patients with CF.

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