Abstract: Objective To explore the clinical significance of rapid diagnosis of fungal otitis externa using fluorescence staining method. Method: A retrospective analysis was conducted on 30 children with suspected fungal otitis externa who were diagnosed at the Otolaryngology Outpatient Department of Xiamen Children's Hospital from January 2021 to December 2023. Each case underwent external ear canal secretion fluorescence staining and fungal culture, and the positive rates of fungal infection detected by fluorescence staining and fungal culture were compared. Result: Among the 30 suspected children, 28 cases were detected as fungal infections by fluorescence staining, and 26 cases were detected as fungal infections by fungal culture. The positive rate of fluorescence staining for fungal otitis externa in children (93.3%) was higher than that of fungal culture (86.7%), and the difference was statistically significant（ χ 2=0.04, P<0.05). Conclusion: The fluorescence staining method has high sensitivity in diagnosing fungal otitis externa, and the operation is simpler and faster, which is worthy of widespread clinical promotion.