Objective To explore the clinical application effect of rapid diagnostic protocol for superficial cutaneous fungal infections based on fluorescent staining method. Methods A total of 90 patients with superficial cutaneous fungal infections admitted to Guangzhou Dermatology Hospital from January 2024 to December 2025 were selected and received rapid histological diagnosis. The diagnostic performance of potassium hydroxide wet mount method and fluorescent staining method was compared. Results The positive detection rates of fluorescent staining method for lesions on head and face, hands and feet, nail lesions and the overall detection rate were higher than those of potassium hydroxide wet mount method (P <0.05). The positive detection rates for onychomycosis, tinea corporis, tinea manus and pedis, tinea versicolor, Malassezia folliculitis and the overall detection rate of fluorescent staining method were higher than those of potassium hydroxide wet mount method (P <0.05). Conclusion Compared with potassium hydroxide wet mount method, fluorescent staining method has definite efficacy in the rapid diagnosis of superficial cutaneous fungal infections, with high positive detection rate for lesions of different types and sites. It can provide objective evidence for the formulation of clinical diagnosis and treatment schemes.