Optimal Susceptibility Testing Conditions for Detection of Azole Resistance in Aspergillus spp.: NCCLS Collaborative Evaluation
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The most important role of susceptibility testing is to identify potentially resistant isolates for the agent being evaluated. Standard testing guidelines recently have been proposed for antifungal susceptibility testing of filamentous fungi (molds). This collaborative (eight centers) study evaluated further newly proposed guidelines (NCCLS, proposed standard M38-P, 1998) and other testing conditions for antifungal susceptibility testing of Aspergillus spp. to itraconazole and three new triazoles, posaconazole (SCH56592), ravuconazole (BMS-207147), and voriconazole. MICs of itraconazole, posaconazole, ravuconazole, and voriconazole for 15 selected isolates of three species of Aspergillus ( A. fumigatus, A. flavus , and A. terreus ) with well documented in vitro, clinical, or animal data were determined in each center by using four medium formulations (standard RPMI-1640 [RPMI], RPMI with 2% dextrose, antibiotic medium 3 [M3], and M3 with 2% dextrose) and two criteria of MIC determination (complete [MIC-0s] and prominent [MIC-2s] growth inhibition) at 24, 48, and 72 h. The highest reproducibility (92 to 99%) was seen with the standard RPMI and M3 media. Moreover, the distinction between itraconazole-resistant (MICs of >8 μg/ml for clinically resistant strains) and -susceptible (MICs of 0.03 to 1 μg/ml) isolates, as well as between a voriconazole-resistant laboratory mutant and other isolates (voriconazole MICs of 2 to >8 versus 0.12 to 2 μg/ml), was more consistently evident with the standard RPMI medium and when MIC-0s were determined at 48 h. These results provide further refinement of the testing guidelines for susceptibility testing of Aspergillus spp. and warrant consideration for inclusion in the future NCCLS document M38-A.
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For the five molds and the yeast isolate, >99% of the inocula were within the target ranges (0.4 × 10^6 to 5 × 10^6 CFU/ml for molds and 1 × 10^6 to 5 × 10^6 CFU/ml for C. krusei).
“for the five molds and the yeast isolate, >99% of the inocula were within the target ranges (0.4 × 106 to 5 × 106 CFU/ml [molds] and 1 × 106 to 5 × 106 CFU/ml[C. krusei]) or similar reproducibility as in previous collaborative studies (92 to 95%) (1, 2, 7, 8).”
From Quality Control Guidelines for Amphotericin B, Itraconazole, Posaconazole, and Voriconazole Disk Diffusion Susceptibility Tests with Nonsupplemented Mueller-Hinton Agar (CLSI M51-A Document) for Nondermatophyte Filamentous Fungi (2011) · cites this paper as [8]
The Materials and Methods (page unavailable) give the stated target range for mold inocula and report that 95% of measured stock inocula fell within a narrower range. They do not report >99% within the target range for five molds and C. krusei; the study evaluated three Aspergillus species, while C. krusei is mentioned only in preliminary quality-control testing. (LLM, conf=0.86)
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Espinel-Ingroff, A., Canton, E., Fothergill, A., Ghannoum, M., Johnson, E., Jones, R. N., Ostrosky-Zeichner, L., Schell, W., Gibbs, D. L., Wang, A., et al. (2011) · Journal of Clinical Microbiology · doi:10.1128/jcm.00393-11 · Cited by 78 · see citing works
Self-citation
- Authors
- Espinel-Ingroff, A., Bartlett, M., Chaturvedi, V., Ghannoum, M., Hazen, K. C., Pfaller, M. A., Rinaldi, M., & Walsh, T. J.
- Affiliations
- <!--label omitted: 1-->Medical College of Virginia of Virginia Commonwealth University, Richmond,1 and
- Indiana University School of Medicine (US)
- New York State Department of Health (US)
- University Hospitals of Cleveland (US)
- University of Virginia Health System (US)
- University of Iowa (US)
- The University of Texas at San Antonio Health Science Center (US)
- National Cancer Institute (US)
- Venue
- Antimicrobial Agents and Chemotherapy
- Publisher
- American Society for Microbiology
- Published
- 2001-06
- Volume
- 45
- Issue
- 6
- Pages
- 1828-1835
- ISSN
- 0066-4804 (ISSN-L)
- 1098-6596
- PMC
- PMC90553
- OpenAlex
- W2163900733
- Cited by
- 83
- References
- 3
- Citations by year
- 2026: 1 · 2022: 1 · 2020: 1 · 2019: 2 · 2018: 3 · 2017: 2 · 2016: 4 · 2015: 3 · 2012: 3
- FWCI
- 7.20
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Antimicrobial Agents and Chemotherapy (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Pfizer
- Topics
- Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Bacterial Identification and Susceptibility Testing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Keywords
- antifungal susceptibility testing, Aspergillus, azole resistance, itraconazole, voriconazole, posaconazole, ravuconazole, triazoles, minimum inhibitory concentration, RPMI medium, dextrose supplementation, reproducibility, Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, filamentous fungi
- MeSH terms
- Antifungal Agents, Aspergillus, Microbial Sensitivity Tests, Triazoles
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