Characterization of Heteroresistance to Fluconazole among Clinical Isolates of Cryptococcus neoformans
Open accessAbstract
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.
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C. neoformans heteroresistance was detected at 35 °C but not at 30 °C.
“For example it was found that C. neoformans heteroresistance was detected at 35 °C but not at 30 °C, indicating the importance of environmental temperature [19].”
From Insight into the Mechanisms and Clinical Relevance of Antifungal Heteroresistance (2025) · cites this paper as [19]
The Results state the opposite of the cited claim: heteroresistance was absent at 35°C and demonstrable only at 30°C in agar-based tests. The paper's opening summary reports the same temperature dependence. (LLM, conf=0.95)
Checked by a reasoning model — may be wrong.
Experimental — accuracy not yet measuredEnvironmental temperature is important for C. neoformans heteroresistance.
“For example it was found that C. neoformans heteroresistance was detected at 35 °C but not at 30 °C, indicating the importance of environmental temperature [19].”
From Insight into the Mechanisms and Clinical Relevance of Antifungal Heteroresistance (2025) · cites this paper as [19]
The Results section reports the opposite temperature pattern: heteroresistance was absent at 35°C and demonstrable only at 30°C in agar-based tests. The Discussion confirms that incubation temperature affected the phenotype, but the citing sentence reverses the study’s finding. (LLM, conf=0.90)
Checked by a reasoning model — may be wrong.
Experimental — accuracy not yet measuredFor C. neoformans and C. gattii, heteroresistance to fluconazole is intrinsic and increased resistance can be induced.
“For C. neoformans and C. gattii, the heteroresistance to fluconazole is intrinsic and increased resistance can be induced [19,102].”
From Insight into the Mechanisms and Clinical Relevance of Antifungal Heteroresistance (2025) · cites this paper as [19]
The Results identify pre-existing fluconazole-heteroresistant subpopulations in 5 of 107 C. neoformans isolates, and the Discussion reports increased resistance after fluconazole exposure in vitro. The paper does not establish that heteroresistance is intrinsic to C. neoformans generally, and it does not address C. gattii. (LLM, conf=0.88)
Experimental — accuracy not yet measured
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- Authors
- Yamazumi, T., Pfaller, M. A., Messer, S. A., Houston, A. K., Boyken, L., Hollis, R. J., Furuta, I., & Jones, R. N.
- Affiliations
- University of Iowa (US)
- Kindai University (JP)
- Tufts University (US)
- Venue
- Journal of Clinical Microbiology
- Publisher
- American Society for Microbiology
- Published
- 2003-01
- Volume
- 41
- Issue
- 1
- Pages
- 267-272
- ISSN
- 0095-1137 (ISSN-L)
- 1098-660X
- PMC
- PMC149577
- OpenAlex
- W2134155337
- Cited by
- 62
- References
- 31
- Citations by year
- 2026: 2 · 2025: 5 · 2024: 1 · 2023: 2 · 2022: 6 · 2021: 1 · 2020: 2 · 2019: 3 · 2018: 2 · 2017: 3 · 2016: 4 · 2015: 1 · 2014: 2 · 2013: 5 · 2012: 2
- FWCI
- 1.41
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Journal of Clinical Microbiology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Centers for Disease Control and Prevention
- Pfizer Pharmaceuticals
- Topics
- Health Sciences › Medicine › Epidemiology › Fungal Infections and Studies
- Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
- Life Sciences › Agricultural and Biological Sciences › Ecology, Evolution, Behavior and Systematics › Fungal Plant Pathogen Control
- Keywords
- Cryptococcus neoformans, fluconazole, heteroresistance, antifungal resistance, clinical isolates, minimum inhibitory concentration, subclonal selection, incubation temperature
- MeSH terms
- Antifungal Agents, Cryptococcosis, Cryptococcus neoformans, Humans, Microbial Sensitivity Tests, Temperature, Fluconazole, Prevalence, Drug Resistance, Fungal
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