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Journal of Clinical Microbiology, August 2002, p. 2772-2778, Vol. 40, No. 8
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.8.2772-2778.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Genetic Analysis of Pseudomonas aeruginosa Isolates from the Sputa of Australian Adult Cystic Fibrosis Patients

Mario Anthony,1 Barbara Rose,1 Mary Beard Pegler,1 Mark Elkins,2 Helen Service,1 Keerthi Thamotharampillai,1 Jason Watson,1 Michael Robinson,2 Peter Bye,2 John Merlino,1 and Colin Harbour1*

Department of Infectious Diseases, Faculty of Medicine, University of Sydney,1 Department of Respiratory Medicine, Royal Prince Alfred Hospital, Sydney, Australia2

Received 4 January 2002/ Returned for modification 21 April 2002/ Accepted 18 May 2002

Genetic investigations were carried out with 50 phenotypically selected strains of Pseudomonas aeruginosa from 18 patients attending an Australian cystic fibrosis (CF) center. The isolates were analyzed by restriction fragment length polymorphism (RFLP) analysis by pulsed-field gel electrophoresis (PFGE). Phylogenetic analysis of the macrorestriction patterns showed rates of genetic similarity ranging from 76 to 100%; 24 (48%) of the strains from 11 patients had greater than 90% similarity. A dominant strain emerged: 15 isolates from seven patients had identical PFGE patterns, and 4 other isolates were very closely related. The 50 isolates were grouped into 21 pulsotypes on the basis of visual delineation of a three-band difference. Ten of the 18 (56%) patients were infected with clonal or subclonal strains. Sequence analysis of PCR products derived from the mucA gene showed 20 mutations, with the number of mutations in individual isolates ranging from 1 to 4; 19 of these changes are reported here for the first time. Potentially functional changes were found in 22 (44%) isolates. Eight changes (five transversions and three single base deletions) led to premature stop codons, providing support for the presence of mucA mutations as one pathway to mucoidy. There was a trend toward an association between the dominant strain and lack of potentially functional mucA mutations (P = 0.09 by the {chi}2 test) but no relationship between genotype and phenotype. This is the first study of genetic variation in P. aeruginosa isolates from adult Australian CF patients. The findings highlight the need for further investigations on the transmissibility of P. aeruginosa in CF patients.

* Corresponding author. Mailing address: Department of Infectious Diseases, Blackburn Bldg., D06, University of Sydney, Sydney, NSW, 2006, Australia. Phone: 61 02 93514334. Fax: 61 02 93514731. E-mail: charbour{at}infdis.usyd.edu.au.

Journal of Clinical Microbiology, August 2002, p. 2772-2778, Vol. 40, No. 8
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.8.2772-2778.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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