A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing
| Autor: | Jose L. Lopez-Ribot, Priya Uppuluri, Christopher G. Pierce, Amanda R Tristan, Gordon Ramage, Floyd L. Wormley, Eilidh Mowat |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Antifungal
Susceptibility testing Formazans medicine.drug_class Cell Culture Techniques Fungi Biofilm Biology Plant biology Article General Biochemistry Genetics and Molecular Biology Microbiology chemistry.chemical_compound Plant science Science research Biochemistry chemistry Drug Resistance Fungal Biofilms medicine Formazan 96 well plate |
| Zdroj: | Nature Protocols. 3:1494-1500 |
| ISSN: | 1750-2799 1754-2189 |
| Popis: | The incidence of fungal infections has increased significantly over the past decades. Very often these infections are associated with biofilm formation on implanted biomaterials and/or host surfaces. This has important clinical implications, as fungal biofilms display properties that are dramatically different from planktonic (free-living) populations, including increased resistance to antifungal agents. Here we describe a rapid and highly reproducible 96-well microtiter-based method for the formation of fungal biofilms, which is easily adaptable for antifungal susceptibility testing. This model is based on the ability of metabolically active sessile cells to reduce a tetrazolium salt (2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide) to water-soluble orange formazan compounds, the intensity of which can then be determined using a microtiter-plate reader. The entire procedure takes approximately 2 d to complete. This technique simplifies biofilm formation and quantification, making it more reliable and comparable among different laboratories, a necessary step toward the standardization of antifungal susceptibility testing of biofilms. |
| Databáze: | OpenAIRE |
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