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Journal of Clinical Microbiology, January 2003, p. 403-409, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.403-409.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Evaluation of Broth Microdilution Testing Parameters and Agar Diffusion Etest Procedure for Testing Susceptibilities of Aspergillus spp. to Caspofungin Acetate (MK-0991)

Ana Espinel-Ingroff*

Medical College of Virginia Campus, Virginia Commonwealth University, Richmond, Virginia 23298

Received 21 August 2002/ Returned for modification 24 September 2002/ Accepted 14 October 2002

The NCCLS M38-A document does not describe guidelines for testing caspofungin acetate (MK-0991) and other echinocandins against molds. This study evaluated the susceptibilities of 200 isolates of Aspergillus fumigatus, A. flavus, A. nidulans, A. niger, and A. terreus to caspofungin (MICs and minimum effective concentrations [MECs]) by using standard RPMI 1640 (RPMI) and antibiotic medium 3 (M3), two inoculum sizes (103 and 104 CFU/ml), and two MIC determination criteria (complete [MICs-0] and prominent growth inhibition [MICs-2]) at 24 and 48 h. Etest MICs were also determined. In general, caspofungin MIC-2 and MEC pairs were comparable with both media and inocula (geometric mean ranges of MECs and MICs, respectively, with larger inoculum: 0.12 to 0.64 µg/ml and 0.12 to 0.44 µg/ml with RPMI versus 0.04 to 0.51 µg/ml and 0.03 to 0.21 µg/ml with M3); however, MEC results were less influenced by testing conditions than MICs, especially with the larger inoculum. Overall, the agreement between caspofungin Etest MICs and broth dilution values was higher with MECs obtained with M3 (>90%) and the large inoculum than under the other testing conditions. Because RPMI is a more stable and chemically defined medium than M3, the determination at 24 h of the easier visual MECs with RPMI and the inoculum recommended in the M38-A document appears to be a suitable procedure at present for in vitro testing of caspofungin against Aspergillus spp. Future in vitro correlations with in vivo outcome of both microdilution and Etest procedures may detect more-relevant testing conditions.

* Mailing address: Division of Infectious Diseases, Medical Mycology Research Laboratory, Medical College of Virginia Campus/VCU, Richmond, VA 23398. Phone (804) 828-9711. Fax (804) 828-3097. E-mail: avingrof{at}hsc.vcu.edu.

Journal of Clinical Microbiology, January 2003, p. 403-409, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.403-409.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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