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Journal of Clinical Microbiology, September 2003, p. 4252-4258, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4252-4258.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Modification of Rapid Susceptibility Assay for Antifungal Susceptibility Testing of Aspergillus fumigatus

Tracy J. Wetter,1,3* Kevin C. Hazen,2 and Jim E. Cutler3

Department of Microbiology, Montana State University, Bozeman, Montana 59718,1 Department of Pathology, University of Virginia Health System, Charlottesville, Virginia 22908,2 Research Institute for Children, Children's Hospital, New Orleans, Louisiana 701183

Received 3 March 2003/ Returned for modification 7 April 2003/ Accepted 2 June 2003

To improve objectivity and speed of current antifungal mold susceptibility testing, the yeast Rapid Susceptibility Assay (RSA) was adapted for Aspergillus species. The RSA is based on glucose utilization in the presence of an antifungal drug. Aspergillus fumigatus conidia were incubated in 0.2% glucose RPMI 1640 containing 0.03 to 16 µg of amphotericin B or itraconazole/ml. Drug-related inhibition of glucose utilization correlated with suppression of conidial germination. Following incubation of conidia with various concentrations of antifungal drug, the percentage of residual glucose in the growth medium was determined colorimetrically and plotted against drug concentration to determine the MIC (MICRSA). National Committee for Clinical Laboratory Standards (NCCLS) M38-P testing was also performed to obtain NCCLS MICs (MICNCCLS) for direct comparison with MICRSAs. Conidial inocula of an optical density at 530 nm (OD530) of 0.11 facilitated determination of amphotericin B and itraconazole MICRSAs at 16 h equal to or within a single twofold dilution of MICNCCLSs obtained at 48 h. Preliminary testing with a 0.11-OD530 conidial inoculum of the slower-growing Aspergillus terreus resulted in itraconazole and amphotericin B MICRSAs at 16 h equal to or within a single twofold dilution of MICNCCLSs obtained at 48 h. These data indicate that the mold RSA provides a more objective and rapid method for Aspergillus spp. susceptibility testing than the NCCLS M38-P assay.

* Corresponding author. Mailing address: Research Institute for Children, 200 Henry Clay Ave., New Orleans, LA 70118. Phone: (504) 896-2712. E-mail: twetter{at}chnola-research.org.

Journal of Clinical Microbiology, September 2003, p. 4252-4258, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4252-4258.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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