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Journal of Clinical Microbiology, November 2009, p. 3444-3448, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.00459-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Cross-Sectional and Longitudinal Multilocus Sequence Typing of Pseudomonas aeruginosa in Cystic Fibrosis Sputum Samples{triangledown}

David J. Waine,1,2,3* David Honeybourne,1,2 E. Grace Smith,4 Joanna L. Whitehouse,2 and Chris G. Dowson1

Department of Biological Sciences, Warwick University, Coventry, CV4 7AL, United Kingdom,1 West Midlands Adult CF Unit, Heart of England Foundation Trust, Birmingham, B9 5SS, United Kingdom,2 Chest Clinic, Derriford Hospital, Plymouth, PL6 8DH, United Kingdom,3 Department of Microbiology, Heart of England Foundation Trust, Birmingham, B9 5SS, United Kingdom4

Received 4 March 2009/ Returned for modification 9 April 2009/ Accepted 19 August 2009

Multilocus sequence typing (MLST) is a genetic typing tool designed to provide information about the relatedness of isolates at the core genome level. The utility of MLST in regard to cystic fibrosis (CF)-related infection with Pseudomonas aeruginosa is unknown. The molecular clock speed of the MLST genes was studied using 219 colonies isolated longitudinally from 49 patients with CF. A cross-sectional study examining 27 to 46 colonies per sputum sample for samples from 16 patients was also undertaken. The molecular clock speed was estimated to be 2.05 x 10–5 (upper 95% confidence limit) or 4.75 x 10–6 (50% confidence limit) point mutations per nucleotide per year. In the cross-sectional study, 50% of patients were infected with more than one sequence type. There was evidence of point mutations, recombination events, and coinfection with epidemic and unique strains. A clonal complex that was highly genetically distinct from the rest of the P. aeruginosa population was identified. The MLST scheme uses genes with an appropriate clock speed and provides useful information about the genetic variation of P. aeruginosa within and between patients with CF.

* Corresponding author. Mailing address: Chest Clinic, Derriford Hospital, Plymouth, PL6 8DH, United Kingdom. Phone: 44 (0)1752 431490. Fax: 44 (0)1752 763864. E-mail: David.Waine{at}phnt.swest.nhs.uk

{triangledown} Published ahead of print on 26 August 2009.

Journal of Clinical Microbiology, November 2009, p. 3444-3448, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.00459-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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