Use of turbidimetric growth curves for early determination of antifungal drug resistance of filamentous fungi.
SourceJournal of Clinical Microbiology, 41, 10, (2003), pp. 4718-4725
Article / Letter to editor
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Journal of Clinical Microbiology
SubjectUMCN 4.1: Microbial pathogenesis and host defense
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.
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