Cited by 144Antifungal resistance and susceptibilityMicrobial Natural Products and BiosynthesisEssential Oils and Antimicrobial Activity

A New Approach to Drug Discovery: High-Throughput Screening of Microbial Natural Extracts against Aspergillus fumigatus Using Resazurin

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Monteiro, Maria Cândida, de la Cruz, Mercedes, Cantizani, Juan, Moreno, Catalina, Tormo, José R., Mellado, Emilia, De Lucas, J. Ramón, Asensio, Francisco +5 more·2012·SLAS Discovery

Abstract

Natural products are an inexhaustible source for drug discovery. However, the validation and selection of primary screening assays are vital to guarantee a selection of extracts or molecules with relevant pharmacological action and worthy of following up. The assay must be rapid, simple, easy to implement, and produce quick results and preferably at a low cost. In this work, we developed and validated a colorimetric microtiter assay using the resazurin viability dye. The parameters of the resazurin method for high-throughput screening (HTS) using natural extracts against Aspergillus fumigatus were optimized and set up. The extracts plus RPMI-1640 modified medium containing the spores and 0.002% resazurin were added per well. The fluorescence was read after 24 to 30 h of incubation. The resazurin proved to be as suitable as Alamar Blue for determining the minimal inhibitory concentration of different antifungals against A. fumigatus and effective to analyze fungicidal and fungistatic compounds. An HTS of 12 000 microbial extracts was carried out against two A. fumigatus strains, and 2.7% of the extracts displayed antifungal activity. Our group has been the first to use this methodology for screening a collection of natural extracts to identify compounds with antifungal activity against the medically important human pathogen A. fumigatus.

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Crossref · 10.1177/1087057111433459

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15 of 15 references verified0 citation errors · 0 not checkable

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  • Tetrazolium salts, MTT and XTT are often used in MIC endpoint determination for both antifungal and antibacterial microdilution assays.

    “Tetrazolium salts, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 2,3-bis {2-methoxy-4-nitro-5-[(sulfenylamino) carbonyl]-2H-tetrazolium-hydroxide} (XTT), are often used in the MIC endpoint determination for both antifungal and antibacterial microdilution assays [57], [58], [59], [60].”

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Authors
Monteiro, M. C., de la Cruz, M., Cantizani, J., Moreno, C., Tormo, J. R., Mellado, E., De Lucas, J. R., Asensio, F., Valiante, V., Brakhage, A. A., Latgé, J.-P., Genilloud, O., & Vicente, F.
Affiliations
  • Fundación Medina (ES)
  • Instituto de Salud Carlos III (ES)
  • Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie e. V. - Hans-Knöll-Institut (HKI) (DE)
  • Institut Pasteur (FR)
Corresponding author
Monteiro, Maria Cândida
Venue
SLAS Discovery
Publisher
Elsevier BV
Published
2012-04
Volume
17
Issue
4
Pages
542-549
ISSN
2472-5552 (ISSN-L)
OpenAlex
W2012818739
Cited by
144
References
15
Citations by year
2026: 4 · 2025: 6 · 2024: 17 · 2023: 17 · 2022: 11 · 2021: 19 · 2020: 16 · 2019: 6 · 2018: 11 · 2017: 7 · 2016: 8 · 2015: 9 · 2014: 7 · 2013: 6
FWCI
4.88
Type
journal article
Language
en
Open access
Locations
  • SLAS DISCOVERY (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • Universidad de Granada
  • Fundación MEDINA
  • Junta de Andalucía
Topics
  • Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
  • Health Sciences › Medicine › Pharmacology › Microbial Natural Products and Biosynthesis
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Essential Oils and Antimicrobial Activity
Keywords
Aspergillus fumigatus, resazurin, high-throughput screening, antifungal activity, minimum inhibitory concentration, drug discovery
MeSH terms
Antifungal Agents, Aspergillus fumigatus, Fungi, Humans, Indicators and Reagents, Microbial Sensitivity Tests, Oxazines, Xanthenes, Complex Mixtures, Drug Discovery, High-Throughput Screening Assays

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