Influence of incubation time, inoculum size, and glucose concentrations on spectrophotometric endpoint determinations for amphotericin B, fluconazole, and itraconazole

Nguyen, M. Hong; Yu, Christine Y.

doi:10.1128/jcm.37.1.141-145.1999

Cited by 48Antifungal resistance and susceptibilityBacterial Identification and Susceptibility TestingFungal Infections and Studies

Influence of incubation time, inoculum size, and glucose concentrations on spectrophotometric endpoint determinations for amphotericin B, fluconazole, and itraconazole

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Nguyen, M. Hong, Yu, Christine Y.·1999·Journal of Clinical Microbiology

DOI 10.1128/jcm.37.1.141-145.1999PMC PMC84190

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Abstract

We addressed the influence of the incubation time (24 h versus 48 h), starting inoculum size (standard inoculum size, approximately 10(3) CFU/ml, versus large inoculum size, approximately 10(4) CFU/ml), and supplementation with 2% glucose of RPMI 1640 medium on the spectrophotometric determination of the MICs of amphotericin B, fluconazole, and itraconazole. We compared the MICs determined spectrophotometrically with those determined by the standard broth macrodilution method (National Committee for Clinical Laboratory Standards approved guideline M27-A). The agreement between the results of the spectrophotometric and standard methods for amphotericin B testing was 100%; this agreement was independent of the inoculum size and incubation time. On the other hand, the agreement for the results for fluconazole testing and itraconazole testing was dependent on the inoculum size and incubation time. With large inoculum size, excellent agreement can be achieved at 24 h. With standard inoculum size, acceptable agreement can be achieved only at 48 h. In contrast to previous observations, the addition of 2% glucose did not have any significant impact on the growth density at 24 h, nor did it improve the agreement with the standard method. Furthermore, supplemental glucose might falsely elevate the MIC at 48 h.

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Inoculum size can affect susceptibility results.
“It is well recognized that the inoculum size can affect susceptibility results [130,144,145,146,147].”
From Candida glabrata Biofilms: How Far Have We Come? (2017) · cites this paper as [144]

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Authors: Nguyen, M. H., & Yu, C. Y.
Affiliations: University of Florida Health (US)
ORCID: Nguyen, M. Hong
Yu, Christine Y.
Venue: Journal of Clinical Microbiology
Publisher: American Society for Microbiology
Published: 1999-01
Volume: 37
Issue: 1
Pages: 141-145
ISSN: 0095-1137 (ISSN-L)
1098-660X
DOI: 10.1128/jcm.37.1.141-145.1999
PMC: PMC84190
OpenAlex: W2968724175
Cited by: 47
References: 18
Citations by year: 2025: 1 · 2024: 1 · 2023: 3 · 2022: 1 · 2021: 1 · 2018: 3 · 2017: 2 · 2015: 2 · 2014: 1 · 2013: 2 · 2012: 2
FWCI: 2.97
Type: journal article
Language: en
Open access: green
europepmc.org
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
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
Health Sciences › Medicine › Epidemiology › Fungal Infections and Studies
Keywords: minimum inhibitory concentration, amphotericin B, fluconazole, itraconazole, incubation time, inoculum size, glucose supplementation, RPMI 1640, broth macrodilution, antifungal susceptibility testing, Candida
MeSH terms: Amphotericin B, Antifungal Agents, Candida, Culture Media, Glucose, Humans, Microbial Sensitivity Tests, Spectrophotometry, Time Factors, Fluconazole, Itraconazole

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Influence of incubation time, inoculum size, and glucose concentrations on spectrophotometric endpoint determinations for amphotericin B, fluconazole, and itraconazole

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