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Antimicrobial Agents and Chemotherapy, June 2001, p. 1828-1835, Vol. 45, No. 6
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.6.1828-1835.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Optimal Susceptibility Testing Conditions for Detection of Azole Resistance in Aspergillus spp.: NCCLS Collaborative Evaluation

A. Espinel-Ingroff,1,* M. Bartlett,2 V. Chaturvedi,3 M. Ghannoum,4 K. C. Hazen,5 M. A. Pfaller,6 M. Rinaldi,7 and T. J. Walsh8

Medical College of Virginia of Virginia Commonwealth University, Richmond,1 and University of Virginia Health System, Charlottesville,5 Virginia; Indiana University School of Medicine, Indianapolis, Indiana2; New York Health Department, Albany, New York3; University Hospital of Cleveland, Case Western Reserve University, Cleveland, Ohio4; University of Iowa College of Medicine, Iowa City, Iowa6; University of Texas Health Science Center, San Antonio, Texas7; and National Cancer Institute, Bethesda, Maryland8

Received 13 October 2000/Returned for modification 6 December 2000/Accepted 27 March 2001

The most important role of susceptibility testing is to identify potentially resistant isolates for the agent being evaluated. Standard testing guidelines recently have been proposed for antifungal susceptibility testing of filamentous fungi (molds). This collaborative (eight centers) study evaluated further newly proposed guidelines (NCCLS, proposed standard M38-P, 1998) and other testing conditions for antifungal susceptibility testing of Aspergillus spp. to itraconazole and three new triazoles, posaconazole (SCH56592), ravuconazole (BMS-207147), and voriconazole. MICs of itraconazole, posaconazole, ravuconazole, and voriconazole for 15 selected isolates of three species of Aspergillus (A. fumigatus, A. flavus, and A. terreus) with well documented in vitro, clinical, or animal data were determined in each center by using four medium formulations (standard RPMI-1640 [RPMI], RPMI with 2% dextrose, antibiotic medium 3 [M3], and M3 with 2% dextrose) and two criteria of MIC determination (complete [MIC-0s] and prominent [MIC-2s] growth inhibition) at 24, 48, and 72 h. The highest reproducibility (92 to 99%) was seen with the standard RPMI and M3 media. Moreover, the distinction between itraconazole-resistant (MICs of >8 µg/ml for clinically resistant strains) and -susceptible (MICs of 0.03 to 1 µg/ml) isolates, as well as between a voriconazole-resistant laboratory mutant and other isolates (voriconazole MICs of 2 to >8 versus 0.12 to 2 µg/ml), was more consistently evident with the standard RPMI medium and when MIC-0s were determined at 48 h. These results provide further refinement of the testing guidelines for susceptibility testing of Aspergillus spp. and warrant consideration for inclusion in the future NCCLS document M38-A.

* Corresponding author. Mailing address: Medical Mycology Research Laboratory, Medical College of Virginia/VCU, P.O. Box 980049, 1101 E. Marshall St., Sanger Hall, Room 7-049, Richmond, VA 23298-0049. Phone: (804) 828-9711. Fax: (804) 828-3097. E-mail: avingrof{at}hsc.vcu.edu.

Antimicrobial Agents and Chemotherapy, June 2001, p. 1828-1835, Vol. 45, No. 6
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.6.1828-1835.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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