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Journal of Clinical Microbiology, May 2004, p. 1915-1922, Vol. 42, No. 5
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.5.1915-1922.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Clinically Feasible Biofilm Susceptibility Assay for Isolates of Pseudomonas aeruginosa from Patients with Cystic Fibrosis

Samuel M. Moskowitz,1,2 Jessica M. Foster,3 Julia Emerson,1,2 and Jane L. Burns1,3*

Department of Pediatrics, University of Washington School of Medicine, Seattle, Washington 98195,1 Divisions of Pulmonary Medicine,2 Infectious Diseases, Children's Hospital and Regional Medical Center, Seattle, Washington 981053

Received 26 September 2003/ Returned for modification 27 October 2003/ Accepted 6 February 2004

Pseudomonas aeruginosa is the predominant cause of chronic airway infection in cystic fibrosis (CF). CF airway isolates are often tested for antibiotic susceptibility but are rarely eradicated by the antibiotics identified as potentially effective. The growth state of P. aeruginosa in CF airways is probably different from that exhibited under conventional susceptibility testing conditions and may represent a bacterial biofilm. Biofilm susceptibility testing methods were adapted to create an assay for implementation in a clinical microbiology laboratory. This assay gave reproducible results when examined in 300 paired determinations with 12 antimicrobial agents, with a serious error rate of 5.7%. The biofilm assay was used retrospectively to test these 12 agents against 94 isolates from 41 CF patients. The biofilm inhibitory concentrations (BICs) were much higher than the corresponding conventionally determined MICs for the ß-lactam antibiotics (median values: aztreonam, >128 µg/ml versus 4 µg/ml; ceftazidime, 128 µg/ml versus 2 µg/ml; piperacillin-tazobactam, 256 µg/ml versus 4 µg/ml; and ticarcillin-clavulanate, 512 µg/ml versus 16 µg/ml, respectively) and doxycycline (>64 µg/ml versus 16 µg/ml); and similar for meropenem (4 µg/ml versus <= 1 µg/ml), ciprofloxacin (0.5 µg/ml versus 1 µg/ml), and the aminoglycosides amikacin (32 µg/ml versus 16 µg/ml), gentamicin (16 µg/ml versus 8 µg/ml), and tobramycin (4 µg/ml versus 2 µg/ml). The median BIC for azithromycin was 2 µg/ml, whereas isolates were uniformly resistant when tested by standard methods. This demonstrates the feasibility of adapting biofilm susceptibility methods to the clinical microbiology laboratory and opens the way to examining whether biofilm testing might be used to select more effective antibiotic combinations for CF airway infections than methods in current use.

* Corresponding author. Mailing address: Division of Infectious Diseases, Mailstop CW, Children's Hospital and Regional Medical Center, 4800 Sand Point Way, NE, Seattle, WA 98105-0371. Phone: (206) 987-2073. Fax: (206) 987-7311. E-mail: jane.burns{at}seattlechildrens.org.

Journal of Clinical Microbiology, May 2004, p. 1915-1922, Vol. 42, No. 5
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.5.1915-1922.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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