A 96 well microtiter plate-based method for monitoring formation and antifungal susceptibility testing of Candida albicans biofilms.
A 96 well microtiter plate-based method for monitoring formation and antifungal susceptibility testing of Candida albicans biofilms.
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DOI：
10.3791/2287
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发表时间：
2010-10-21
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影响因子：
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通讯作者：
Lopez-Ribot, Jose L
中科院分区：
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作者：
Pierce, Christopher G;Uppuluri, Priya;Lopez-Ribot, Jose L
白色念珠菌仍然是真菌感染的最常见原因，在不断扩大的患者群体中，念珠菌病现在是美国医院第三大常见感染。念珠菌病的不同表现与宿主组织和/或医疗器械（即导管）上的生物膜形成有关。生物膜的形成具有负面的临床意义，因为生物膜内的细胞免受宿主免疫反应和抗真菌药物的作用。我们建立了一种简单、快速、可靠的体外白色念珠菌生物膜形成的96孔滴度板模型，该模型也可用于生物膜抗真菌药敏试验。该试验的读数是比色法，基于代谢活性真菌生物膜细胞对XTT（一种四氮唑盐）的还原。一个典型的实验大约需要24小时来形成生物膜，另外24小时用于抗真菌药敏试验。由于其简单性和使用常见的实验室材料和设备，该技术使生物膜研究大众化，并代表了朝着真菌生物膜抗真菌药敏测试标准化迈出的重要一步。
Candida albicans remains the most frequent cause of fungal infections in an expanding population of compromised patients and candidiasis is now the third most common infection in US hospitals. Different manifestations of candidiasis are associated with biofilm formation, both on host tissues and/or medical devices (i.e. catheters). Biofilm formation carries negative clinical implications, as cells within the biofilms are protected from host immune responses and from the action of antifungals. We have developed a simple, fast and robust in vitro model for the formation of C. albicans biofilms using 96 well microtiter-plates, which can also be used for biofilm antifungal susceptibility testing. The readout of this assay is colorimetric, based on the reduction of XTT (a tetrazolium salt) by metabolically active fungal biofilm cells. A typical experiment takes approximately 24 h for biofilm formation, with an additional 24 h for antifungal susceptibility testing. Because of its simplicity and the use of commonly available laboratory materials and equipment, this technique democratizes biofilm research and represents an important step towards the standardization of antifungal susceptibility testing of fungal biofilms.