Colletotrichum is a relatively uncommon cause of human infection. Previous findings on Colletotrichum keratitis were scarce, and most diagnoses were based on morphological distinction, perhaps underestimating the incidence of Colletotrichum species. This research describes the clinical characteristics and treatment outcomes of 9 cases of Colletotrichum keratitis discovered in our hospital using next-generation sequencing (NGS).We reviewed 78 patients with NGS-proven fungal keratitis between September 1, 2021 and May 31, 2023, 9 patients (11.5 %) were verified as infected with Colletotrichum species, and their medical records were reviewed to identify the clinical characteristics. NGS revealed that 3 patients were infected with C. truncatum, 3 patients with C. gloeosporioides, and the other 3 patients with C. fructicola. Seven patients had a history of corneal plant trauma (all three patients with C. fructicola had corneal injury history due to chestnut burrs), one patient was infected by mosquitoes flying into the eye, and one patient had an unknown origin. Seven patients underwent penetrating keratoplasty, while two patients underwent lamellar keratoplasty. Eight patients healed after keratoplasty, but one required evisceration due to recurrence of fungal infection in the anterior chamber and intractable discomfort.In conclusion NGS allows for more precise diagnosis and enhances epidemiological awareness of Colletotrichum keratitis, which is not as rare as previously reported.