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Journal of Clinical Microbiology, October 2003, p. 4718-4725, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4718-4725.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Use of Turbidimetric Growth Curves for Early Determination of Antifungal Drug Resistance of Filamentous Fungi

Joseph Meletiadis,1,2 Debbie T. A. te Dorsthorst,1,2,3 and Paul E. Verweij1,2*

Department of Medical Microbiology, University Medical Center Nijmegen,1 Department of Medical Microbiology and Infectious Diseases, Canisius-Wilhelmina Hospital,3 Nijmegen University Center for Infectious Diseases, Nijmegen, The Netherlands2

Received 22 January 2003/ Returned for modification 12 March 2003/ Accepted 24 July 2003

A previously described microbroth kinetic system (J. Meletiadis, J. F. Meis, J. W. Mouton, and P. E. Verweij, J. Clin. Microbiol. 39:478-484, 2001) based on continuous monitoring of changes in the optical density of fungal growth was used to describe turbidimetric growth curves of different filamentous fungi in the presence of increasing concentrations of antifungal drugs. Therefore, 24 clinical mold isolates, including Rhizopus oryzae, Aspergillus fumigatus, Aspergillus flavus, and Scedosporium prolificans, were tested against itraconazole, terbinafine, and amphotericin B according to NCCLS guidelines. Among various parameters of the growth curves, the duration of the lag phase was strongly affected by the presence of antifungal drugs. Exposure to increasing drug concentrations resulted in prolonged lag phases of the turbidimetric growth curves. The lag phases of the growth curves at drug concentrations which resulted in more than 50% growth (for itraconazole and terbinafine) and more than 75% growth (for amphotericin B) after 24 h of incubation for R. oryzae, 48 h for Aspergillus spp., and 72 h for S. prolificans were 4 h longer than the lag phases of the growth curves at the corresponding drug-free growth controls which varied from 4.4 h for R. oryzae, 6.5 h for A. flavus, 7.9 h for A. fumigatus, and 11.6 h for S. prolificans. The duration of the lag phases showed small experimental and interstrain variability, with differences of less than 2 h in most of the cases. Using this system, itraconazole and terbinafine resistance (presence of >50% growth) as well as amphotericin B resistance (presence of >75% growth) was determined within incubation periods of 5.0 to 7.7 h for R. oryzae (for amphotericin B resistance incubation for up to 12 h was required), 8.8 to 11.4 h for A. fumigatus, 6.7 to 8.5 h for A. flavus, and 13 to 15.6 h for S. prolificans while awaiting formal MIC determination by the NCCLS reference method.

* Corresponding author. Mailing address: Department of Medical Microbiology, University Medical Center Nijmegen, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands. Phone: 31-24-3619627. Fax: 31-24-3540216. E-mail: p.verweij{at}mmb.umcn.nl.

Journal of Clinical Microbiology, October 2003, p. 4718-4725, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4718-4725.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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