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A New Approach to Drug Discovery: High-Throughput Screening of Microbial Natural Extracts against Aspergillus fumigatus Using Resazurin
Open accessAbstract
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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“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].”
From Methods for in vitro evaluating antimicrobial activity: A review (2016) · cites this paper as [59]
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Self-citation
Metadata
- 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
- ORCID
- 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
- hybrid
- doi.org
- Licence
- 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
- Life Sciences › Immunology and Microbiology › Applied Microbiology and Biotechnology › 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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