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Characterization of Heteroresistance to Fluconazole among Clinical Isolates of Cryptococcus neoformans
Journal article   Open access   Peer reviewed

Characterization of Heteroresistance to Fluconazole among Clinical Isolates of Cryptococcus neoformans

T Yamazumi, M. A Pfaller, S. A Messer, A. K Houston, L Boyken, R. J Hollis, I Furuta and R. N Jones
Journal of clinical microbiology, Vol.41(1), pp.267-272
01/2003
DOI: 10.1128/JCM.41.1.267-272.2003
PMCID: PMC149577
PMID: 12517859
url
https://doi.org/10.1128/JCM.41.1.267-272.2003View
Published (Version of record) Open Access

Abstract

Strains of Cryptococcus neoformans expressing heteroresistance to fluconazole have been described previously. The present study was conducted to investigate the prevalence of heteroresistance among clinical isolates of C. neoformans and to characterize the heteroresistant phenotypes. A total of 107 clinical isolates of C. neoformans for which the MICs of fluconazole ranged from 0.25 to 32 μg/ml were selected. The isolates were chosen to represent a broad geographic distribution. Of the 107 C. neoformans isolates tested, 4 grew on medium containing fluconazole at concentrations that were four to eight times higher than the MICs for each strain. A fifth isolate, for which the fluconazole MIC was 32 μg/ml, grew on agar with 64 μg of fluconazole per ml. These five isolates (4.7% of the total number) were confirmed to exhibit heteroresistant compositions by population analysis. The degree and frequency of resistance varied among the isolates. Stepwise selection by exposure to fluconazole resulted in subclones of all five strains for which the fluconazole MIC was >64 μg/ml. Subclones of three strains demonstrated a homogenous population of resistant cells on medium containing 64 μg of fluconazole/ml. The resistance was sensitive to incubation temperature, that is, heteroresistance was demonstrable only at 30°C by agar-based tests, and was reversible through serial transfers on fluconazole-free medium over a period of 8 days. These results suggest that the fluconazole-heteroresistant phenotype of C. neoformans exists in a significant proportion of clinical isolates and that fluconazole resistance can be developed by selection from heteroresistant clones and induction by exposure to fluconazole.
Mycology

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