title: Rapid genotyping of Pseudomonas aeruginosa isolates harboured by adult and paediatric patients with cystic fibrosis using repetitive-element-based PCR assays.
abstract: 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.
authors.affiliation: Clinical 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.
authors.forename: Melanie W
authors.initials: MW
authors.lastname: Syrmis
authors.affiliation: Clinical 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.
authors.forename: Mark R
authors.initials: MR
authors.lastname: O'Carroll
authors.affiliation: Clinical 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.
authors.forename: Theo P
authors.initials: TP
authors.lastname: Sloots
authors.affiliation: Clinical 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.
authors.forename: Chris
authors.initials: C
authors.lastname: Coulter
authors.affiliation: Clinical 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.
authors.forename: Claire E
authors.initials: CE
authors.lastname: Wainwright
authors.affiliation: Clinical 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.
authors.forename: Scott C
authors.initials: SC
authors.lastname: Bell
authors.affiliation: Clinical 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.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Nissen
mesh_terms: D000293:Adolescent; D000328:Adult; D001315:Australia; D015373:Bacterial Typing Techniques; D002648:Child; D016000:Cluster Analysis; D003550:Cystic Fibrosis / Q000150:complications* / Q000382:microbiology; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D016521:Electrophoresis, Gel, Pulsed-Field; D005260:Female; D005838:Genotype; D006801:Humans; D020071:Interspersed Repetitive Sequences; D008297:Male; D008875:Middle Aged; D017720:Molecular Epidemiology; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011552:Pseudomonas Infections / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial