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Antimicrobial Agents and Chemotherapy, September 2001, p. 2475-2479, Vol. 45, No. 9
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.9.2475-2479.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Standardized Method for In Vitro Antifungal Susceptibility Testing of Candida albicans Biofilms

Gordon Ramage,1 Kacy Vande Walle,2 Brian L. Wickes,1 and José L. López-Ribot2,*

Department of Microbiology1 and Department of Medicine, Division of Infectious Diseases,2 The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78245

Received 29 March 2001/Returned for modification 9 May 2001/Accepted 5 June 2001

Candida albicans is implicated in many biomaterial-related infections. Typically, these infections are associated with biofilm formation. Cells in biofilms display phenotypic traits that are dramatically different from those of their free-floating planktonic counterparts and are notoriously resistant to antimicrobial agents. Consequently, biofilm-related infections are inherently difficult to treat and to fully eradicate with normal treatment regimens. Here, we report a rapid and highly reproducible microtiter-based colorimetric assay for the susceptibility testing of fungal biofilms, based on the measurement of metabolic activities of the sessile cells by using a formazan salt reduction assay. The assay was used for in vitro antifungal susceptibility testing of several C. albicans strains grown as biofilms against amphotericin B and fluconazole and the increased resistance of C. albicans biofilms against these antifungal agents was demonstrated. Because of its simplicity, compatibility with a widely available 96-well microplate platform, high throughput, and automation potential, we believe this assay represents a promising tool for the standardization of in vitro antifungal susceptibility testing of fungal biofilms.

* Corresponding author. Mailing address: Department of Medicine/Division of Infectious Diseases, The University of Texas Health Science Center at San Antonio, South Texas Centers for Biology in Medicine, Texas Research Park, 15355 Lambda Dr., San Antonio, TX 78245. Phone: (210) 562-5017. Fax: (210) 562-5016. E-mail: RIBOT{at}UTHSCSA.EDU.

Antimicrobial Agents and Chemotherapy, September 2001, p. 2475-2479, Vol. 45, No. 9
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.9.2475-2479.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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