A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing

Pierce, C.G., Uppuluri, P., Tristan, A.R., Wormley, F.L., Mowat, E., Ramage, G. and Lopez-Ribot, J.L. (2008) A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nature Protocols, 3(9), pp. 1494-1500. (doi:10.1038/nport.2008.141)

Pierce, C.G., Uppuluri, P., Tristan, A.R., Wormley, F.L., Mowat, E., Ramage, G. and Lopez-Ribot, J.L. (2008) A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nature Protocols, 3(9), pp. 1494-1500. (doi:10.1038/nport.2008.141)

Full text not currently available from Enlighten.

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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Ramage, Professor Gordon
Authors: Pierce, C.G., Uppuluri, P., Tristan, A.R., Wormley, F.L., Mowat, E., Ramage, G., and Lopez-Ribot, J.L.
College/School:College of Medical Veterinary and Life Sciences > School of Medicine, Dentistry & Nursing > Dental School
Journal Name:Nature Protocols
ISSN:1754-2189
ISSN (Online):1750-2799
Published Online:28 August 2008

University Staff: Request a correction | Enlighten Editors: Update this record