Abstract: Objective: To evaluate the effects of fluorescent staining method in detecting skin superficial fungal infection. Methods: Suspected patients with fungal infection were collected in the department of dermatology, and all the samples were detected by fluorescent staining method, KOH wet method and fungal culturing method. Results: The total positivity rate of 236 samples by the fluorescent staining method, KOH wet method and fungal culturing method were 90.6%,67.4% and 60.2% respectively. The positivity rate of 179 lesions by the fluorescent staining method, KOH wet method and fungal culturing method were 92.1%，69.3%，64.2%.The positivity rate of 52 nails were 84.6%，57.7% and 44.2% respectively.. There were significant differences of above positivity rate between fluorescent staining method and KOH wet method or fungal culturing method (s<0.01). The positivity rate of 5 hear samples by the fluorescent staining method, KOH wet method and fungal culturing method were 100%,100% and 80% respectively.There was no significant difference of hear positivity rate among three methods (P>0.05). Conclusions: Fluorescent staining is a simple and effective method ,and it can reduce false negativity rate.