Low Rates of Pseudomonas aeruginosa Misidentification in Isolates from Cystic Fibrosis Patients

Author:

Kidd Timothy J.12,Ramsay Kay A.12,Hu Honghua3,Bye Peter T. P.45,Elkins Mark R.4,Grimwood Keith26,Harbour Colin3,Marks Guy B.5,Nissen Michael D.26,Robinson Philip J.7,Rose Barbara R.3,Sloots Theo P.2,Wainwright Claire E.68,Bell Scott C.19

Affiliation:

1. School of Medicine, University of Queensland, Brisbane, Queensland, Australia

2. Qpid Laboratory, Queensland Children's Medical Research Institute, Royal Children's Hospital and Clinical Medical Virology Centre, University of Queensland, Brisbane, Queensland, Australia

3. Department of Infectious Diseases and Immunology, The University of Sydney, Sydney, New South Wales, Australia

4. Department of Respiratory Medicine, Royal Prince Alfred Hospital, Sydney, New South Wales, Australia

5. The Woolcock Institute of Medical Research, Sydney, New South Wales, Australia

6. Discipline of Paediatrics and Child Health, University of Queensland, Queensland, Australia

7. Department of Respiratory Medicine, Royal Children's Hospital, Melbourne, Victoria, Australia

8. Department of Respiratory Medicine, Royal Children's Hospital and Queensland Children's Medical Research Institute, Brisbane, Queensland, Australia

9. Adult Cystic Fibrosis Centre, Department of Thoracic Medicine, The Prince Charles Hospital, Brisbane, Queensland, Australia

Abstract

ABSTRACT Pseudomonas aeruginosa is an important cause of pulmonary infection in cystic fibrosis (CF). Its correct identification ensures effective patient management and infection control strategies. However, little is known about how often CF sputum isolates are falsely identified as P. aeruginosa . We used P. aeruginosa -specific duplex real-time PCR assays to determine if 2,267 P. aeruginosa sputum isolates from 561 CF patients were correctly identified by 17 Australian clinical microbiology laboratories. Misidentified isolates underwent further phenotypic tests, amplified rRNA gene restriction analysis, and partial 16S rRNA gene sequence analysis. Participating laboratories were surveyed on how they identified P. aeruginosa from CF sputum. Overall, 2,214 (97.7%) isolates from 531 (94.7%) CF patients were correctly identified as P. aeruginosa . Further testing with the API 20NE kit correctly identified only 34 (59%) of the misidentified isolates. Twelve (40%) patients had previously grown the misidentified species in their sputum. Achromobacter xylosoxidans ( n = 21), Stenotrophomonas maltophilia ( n = 15), and Inquilinus limosus ( n = 4) were the species most commonly misidentified as P. aeruginosa . Overall, there were very low rates of P. aeruginosa misidentification among isolates from a broad cross section of Australian CF patients. Additional improvements are possible by undertaking a culture history review, noting colonial morphology, and performing stringent oxidase, DNase, and colistin susceptibility testing for all presumptive P. aeruginosa isolates. Isolates exhibiting atypical phenotypic features should be evaluated further by additional phenotypic or genotypic identification techniques.

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

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