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Antimicrobial Agents and Chemotherapy, June 1999, p. 1383-1386, Vol. 43, No. 6
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The Trailing End Point Phenotype in Antifungal Susceptibility Testing Is pH Dependent

Kieren A. Marr,1,2,* Tige R. Rustad,3 John H. Rex,4 and Theodore C. White3,5

Fred Hutchinson Cancer Research Center,1 Departments of Medicine2 and Pathobiology,3 University of Washington, and Seattle Biomedical Research Institute,5 Seattle, Washington, and Division of Infectious Diseases, Department of Internal Medicine, Center for the Study of Emerging and Reemerging Pathogens, University of Texas Medical School, Houston, Texas4

Received 21 January 1999/Returned for modification 3 March 1999/Accepted 18 March 1999

The interpretation of end points in azole antifungal drug susceptibility testing is problematic, in part due to incomplete growth inhibition of Candida species. Such trailing growth can cause the MICs of fluconazole for some isolates to be low (<1 µg/ml) after 24 h of growth but much higher (>64 µg/ml) after 48 h. Isolates having this type of growth have been described as having a low-high phenotype. Although these isolates would be considered resistant by current National Committee of Clinical Laboratory Standards definitions, growing evidence suggests that they are susceptible in vivo. To further characterize these isolates in vitro, microdilution susceptibility testing comparing the complex defined medium RPMI 1640 to a defined minimal medium (yeast nitrogen broth) was performed. Isolates having trailing growth in MOPS (morpholinepropanesulfonic acid)-buffered RPMI 1640 (pH 7.0) were found to have clear end points in the minimal medium at its native pH of 4.5. The pH of the medium influenced the low-high phenotype, as these same isolates trailed in minimal medium adjusted to a pH of >= 6.0 but did not trail in RPMI 1640 adjusted to a pH of <= 5.0. This pH effect was independent of the medium buffering capacity, as trailing was decreased in both minimal medium and RPMI 1640 (pH 4.5) buffered in citrate. Adjustment in the pH of MOPS-buffered RPMI 1640 reduced trailing in multiple strains of Candida albicans without affecting the MICs for isolates having known susceptible (low-low) and resistant (high-high) phenotypes. Adjustment of the medium pH could be considered to eliminate trailing in azole drug susceptibility testing.

* Corresponding author. Mailing address: Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N. D3-100, Seattle, WA 98109-1024. Phone: (206) 667-2995. Fax: (206) 667-4411. E-mail: Kmarr{at}u.washington.edu.

Antimicrobial Agents and Chemotherapy, June 1999, p. 1383-1386, Vol. 43, No. 6
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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