体外甲真菌病模型的建立及氟康唑甲板内药敏试验

Establishment of the extrinsic onychomycosis model and the drug sensitivity test about fluconazole in the nail

目的建立体外甲真菌病模型并探讨抗真菌药物在甲板内的抗真菌活性。方法建立体外甲真菌病模型:剪取正常人甲,用2.5%NaOH浸泡变软,分别接种红色毛癣菌、石膏样毛癣菌、白色念珠菌和克柔念珠菌,在马铃薯葡萄糖(PD)液体培养基培养30d后取病理切片并行PAS染色,观察各甲板中是否有真菌侵入。甲板内氟康唑药敏试验:根据体外药基法最小抑菌浓度(MIC)值,配制100×MIC、10×MIC、1×MIC3个梯度的10%二甲基亚砜(DMSO)氟康唑透甲液,浸泡软化的甲板,再接种4种致病菌培养,镜下观察甲切片。只有管内全部甲板均未见真菌结构,方判定该菌种对相应浓度下氟康唑敏感。结果在体外甲真菌病模型试验中,甲板角质层遭破坏,可见数目不等的菌丝孢子,形态与甲真菌病病甲切片所见一致,证明离体甲真菌病模型建立。在测定甲板中氟康唑抗真菌活性时,只有石膏样毛癣组菌100×MIC浓度管内甲板均未见真菌结构,将此值与体外所测MIC值对比,二者差异有统计学意义(P<0.05)。结论(1)在正常人离体甲板上接种真菌,可建立体外甲真菌病的感染模型。(2)要达到抑制真菌的作用,甲板内氟康唑浓度必须要高于体外所测MIC值。

Objective To establish extrinsic onychomycosis model and investigate the drug s antifungal effect in the nail.Methods First we established an extrinsic onychomycosis model.The normal nails are destroyed and put into the potato dextrose(PD)fluid medium.Trichophyton rubrum,trichophyton gypseum,candida albicans and candida krusei were cultured with them.After 30 days,the nails were cut into sections and stained through PAS staining method.In the second step,we attempted to testify fluconazole s antifungal abil...