Tampakakis Emmanouil, Okoli Ikechukwu, Mylonakis Eleftherios
Division of Infectious Diseases, Massachusetts General Hospital, Gray-Jackson 504, 55 Fruit Street, Boston, Massachusetts 02114, USA.
Nat Protoc. 2008;3(12):1925-31. doi: 10.1038/nprot.2008.193.
Traditional antimicrobial screens focus on compounds that block the growth of microbial organisms. A new Caenorhabditis elegans-based bioassay can be used for the identification of antifungal compounds that are effective against Candida albicans. According to the protocol, adult nematodes are infected with C. albicans and moved to 96-well plates containing the tested compounds. In the presence of compounds with no antifungal activity, the fungus kills the worms and develops filaments that penetrate through the cuticle. In contrast to traditional screening methods and mammalian models, this facile, time-efficient and less costly assay allows the study of Candida cells in nonplanktonic form and may allow the concurrent evaluation of toxicity and antifungal activity and identify compounds that target virulence factors or modify host immune response. The screening assay takes about 5-6 d depending on the experimental design.
传统的抗菌筛选聚焦于能阻止微生物生长的化合物。一种基于秀丽隐杆线虫的新生物测定法可用于鉴定对白色念珠菌有效的抗真菌化合物。根据该方案，将成年线虫感染白色念珠菌，然后转移至含有受试化合物的96孔板中。在没有抗真菌活性的化合物存在时，真菌会杀死线虫并形成穿透角质层的菌丝。与传统筛选方法和哺乳动物模型不同，这种简便、省时且成本较低的测定法能够研究非浮游形式的念珠菌细胞，还可能同时评估毒性和抗真菌活性，并鉴定靶向毒力因子或改变宿主免疫反应的化合物。根据实验设计，筛选测定大约需要5 - 6天。
