PURPOSE: To analyze the distribution characteristics of ocular fungal isolates and antifungal susceptibility in vitro. DESIGN: A retrospective case-series descriptive study. METHODS: Two thousand one hundred and seventy-nine specimens collected from Tongren Hospital during 2001 to 2004 were identified at the Beijing Institute of Ophthalmology. Fungal culture-positive rate, antifungal susceptibility in vitro, and genus distribution of positive cultures were analyzed retrospectively. For the fungal culture-positive samples, the gender and age of the patients, the location of ocular involvement, and the season of the onset of mycotic ocular diseases were studied. The fungal positive isolates were subjected to antifungal susceptibility testing. RESULTS: Out of 2,179 specimens, 681 specimens were positive cultures. The culture-positive rate was 31.25%. Out of 681 specimens of positive cultures, 591 specimens were from the cornea, 29 from the aqueous humor, 22 from the conjunctiva, 22 from the vitreous body, one from the lacrimal sac, and 16 from the other sites. Fusarium species was the most common pathogen identified in 394 (57.86% of positive cultures), followed by Aspergillus species in 116 (17.03% of positive cultures). Out of 681 positive cultures, the sensitivity in vitro to natamycin, terbinafine, itraconazole, and fluconazole were 608/681 (89.28%), 467/681 (68.58%), 260/681 (38.18%), and 101/681 (14.83%), respectively. Three hundred and sixty-eight (93.4%) of Fusarium species were sensitive to natamycin, 107 (92.2%) of Aspergillus species were sensitive to itraconazole. CONCLUSIONS: Fusaruim species was the predominant pathogen, which resulted in mycotic ocular diseases in northern China, followed by Aspergillus species. Most of the Fusaruim species were sensitive to natamycin, and most of the Aspergillus species were sensitive to itraconazole.