title: A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing.
abstract: 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.
authors.affiliation: Department of Biology and South Texas Center for Emerging Infectious Diseases, The University of Texas at San Antonio, San Antonio, Texas 78249, USA.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Pierce
authors.forename: Priya
authors.initials: P
authors.lastname: Uppuluri
authors.forename: Amanda R
authors.initials: AR
authors.lastname: Tristan
authors.forename: Floyd L
authors.initials: FL
authors.lastname: Wormley
authors.forename: Eilidh
authors.initials: E
authors.lastname: Mowat
authors.forename: Gordon
authors.initials: G
authors.lastname: Ramage
authors.forename: Jose L
authors.initials: JL
authors.lastname: Lopez-Ribot
mesh_terms: D018441:Biofilms / Q000254:growth & development*; D018929:Cell Culture Techniques / Q000379:methods*; D025141:Drug Resistance, Fungal; D005562:Formazans; D005658:Fungi / Q000187:drug effects / Q000254:growth & development*
chemical_list: D005562:Formazans
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