De Brucker Katrijn, De Cremer Kaat, Cammue Bruno P A, Thevissen Karin
Centre for Microbial and Plant Genetics (CMPG), KU Leuven - University of Leuven, Kasteelpark Arenberg 20, Box 2460, Leuven, 3001, Belgium.
Department of Plant Systems Biology, VIB, Technologiepark 927, Ghent, 9052, Belgium.
Methods Mol Biol. 2016;1333:67-72. doi: 10.1007/978-1-4939-2854-5_6.
In contrast to planktonic cultures of the human fungal pathogen Candida albicans, C. albicans biofilms can contain a persister subpopulation that is tolerant to high concentrations of currently used antifungals. In this chapter, the method to determine the persister fraction in a C. albicans biofilm treated with an antifungal compound is described. To this end, a mature biofilm is developed and subsequently treated with a concentration series of the antifungal compound of interest. Upon incubation, the fraction of surviving biofilm cells is determined by plating and plotted versus the used concentrations of the antifungal compound. If a persister subpopulation in the biofilm is present, the dose-dependent killing of the biofilm cells results in a biphasic killing pattern.
与人类真菌病原体白色念珠菌的浮游培养物不同，白色念珠菌生物膜可包含一个对目前使用的抗真菌药物高浓度具有耐受性的持留菌亚群。在本章中，描述了测定用抗真菌化合物处理的白色念珠菌生物膜中持留菌比例的方法。为此，先形成成熟生物膜，随后用一系列感兴趣的抗真菌化合物浓度进行处理。孵育后，通过平板接种确定存活生物膜细胞的比例，并与所使用的抗真菌化合物浓度作图。如果生物膜中存在持留菌亚群，生物膜细胞的剂量依赖性杀伤会导致双相杀伤模式。
