title: A novel 96-well gel-based assay for determining antifungal activity against filamentous fungi.
abstract: In recent years the global rise in antibiotic resistance and environmental consciousness lead to a renewed fervour to find and develop novel antibiotics, including antifungals. However, the influence of the environment on antifungal activity is often disregarded and many in vitro assays may cause the activity of certain antifungals to be overestimated or underestimated. The general antifungal test assays that are economically accessible to the majority of scientists primarily rely on visual examination or on spectrophotometric analysis. The effect of certain morphogenic antifungals, which may lead to hyperbranching of filamentous fungi, unfortunately renders these methods unreliable. To minimise the difficulties experienced as a result of hyperbranching, we developed a straightforward, economical 96-well gel-based method, independent of spectrophotometric analysis, for highly repeatable determination of antifungal activity. For the calculation of inhibition parameters, this method relies on the visualisation of assay results by digitisation. The antifungal activity results from our novel micro-gel dilution assay are comparable to that of the micro-broth dilution assay used as standard reference test of The Clinical and Laboratory Standard Institute. Furthermore, our economical assay is multifunctional as it permits microscopic analysis of the preserved assay results, as well as rendering highly reliable data.
authors.affiliation: BIOPEP Peptide Group, Department of Biochemistry, University of Stellenbosch, Matieland 7600, South Africa.
authors.forename: Anscha Mari
authors.initials: AM
authors.lastname: Troskie
authors.forename: Nicolas Maré
authors.initials: NM
authors.lastname: Vlok
authors.forename: Marina
authors.initials: M
authors.lastname: Rautenbach
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology*; D005658:Fungi / Q000187:drug effects* / Q000254:growth & development; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009181:Mycoses / Q000382:microbiology
chemical_list: D000935:Antifungal Agents