Multicenter evaluation of proposed standardized procedure for antifungal susceptibility testing of filamentous fungi
Open accessAbstract
A multicenter study was conducted to expand the generation and analysis of data that supports the proposal of a reference method for the antifungal susceptibility testing of filamentous fungi. Broth microdilution MICs of amphotericin B and itraconazole were determined in 11 centers against 30 coded duplicate pairs of Aspergillus spp., Fusarium spp., Pseudallescheria boydii, and Rhizopus arrhizus. The effect of inoculum density (approximately 10(3) and 10(4) CFU/ml), incubation time (24, 48, and 72 h), and procedure of MIC determination (conventional and colorimetric [Alamar Blue] evaluation of growth inhibition) on intra- and interlaboratory agreement was analyzed. Based on intra- (97 to 100%) and interlaboratory (94 to 95%) agreement for both drugs, the overall optimal testing conditions identified were determination of colorimetric MICs after 48 to 72 h of incubation with an inoculum density of approximately 10(4) CFU/ml. These testing conditions are proposed as guidelines for a reference broth microdilution method.
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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 [7]
The Results (Table 1) report percentages within the target inoculum range of 87% to 100% across six molds, not >99% for five molds and a yeast isolate. Candida krusei appears in Materials and Methods as a quality-control isolate, but the cited Results do not report a corresponding yeast inoculum percentage. (LLM, conf=0.94)
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- Authors
- Espinel-Ingroff, A., Bartlett, M., Bowden, R., Chin, N. X., Cooper, C., Fothergill, A., McGinnis, M. R., Menezes, P., Messer, S. A., Nelson, P. W., Odds, F. C., Pasarell, L., Peter, J., Pfaller, M. A., Rex, J. H., Rinaldi, M. G., Shankland, G. S., Walsh, T. J., & Weitzman, I.
- Affiliations
- Virginia Commonwealth University (US)
- Johnson & Johnson (United States) (US)
- National Institutes of Health (US)
- Venue
- Journal of Clinical Microbiology
- Publisher
- American Society for Microbiology
- Published
- 1997-01
- Volume
- 35
- Issue
- 1
- Pages
- 139-143
- ISSN
- 0095-1137 (ISSN-L)
- 1098-660X
- PMC
- PMC229526
- OpenAlex
- W2132463351
- Cited by
- 228
- References
- 14
- Citations by year
- 2025: 2 · 2023: 1 · 2021: 1 · 2020: 2 · 2019: 2 · 2018: 3 · 2017: 2 · 2016: 2 · 2015: 6 · 2014: 5 · 2013: 4 · 2012: 4
- FWCI
- 19.89
- 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)
- 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, filamentous fungi, reference method, broth microdilution, minimum inhibitory concentration, amphotericin B, itraconazole, Aspergillus, Fusarium, Pseudallescheria boydii, Rhizopus oryzae, interlaboratory agreement, inoculum size, incubation time, Alamar Blue
- MeSH terms
- Antifungal Agents, Drug Resistance, Microbial, Evaluation Studies as Topic, Fungi, Microbial Sensitivity Tests
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