This study aimed to determine the minimum inhibitory concentrations (MICs) of commonly used antifungal agents against microbial isolates from corneal scrapings, providing valuable insights into the current antifungal efficacy landscape. Corneal scrapings from patients with suspected fungal keratitis were collected and cultured to isolate fungal pathogens. AFST was performed using the broth microdilution method to determine the MICs all total of 5 fungal isolates. MIC50 and MIC90 values were calculated for each antifungal agent against the isolated fungal species. A. Amphotericin B showed the lowest MIC range (0.125-8 μg/ml), with MIC50 and MIC90 values of 1 μg/ml, indicating strong antifungal activity. Itraconazole and ketoconazole had higher MIC values (MIC50 and MIC90 at 32 and 16 μg/ml, respectively), suggesting potential resistance. This study highlights the varying susceptibility of fungal pathogens to commonly used antifungal agents, underscoring the need for routine AFST in managing fungal keratitis. Amphotericin B remains highly effective, while the higher MIC values observed for azoles like itraconazole and ketoconazole suggest emerging resistance. Tailoring antifungal therapy based on susceptibility testing is crucial to improving treatment outcomes in fungal keratitis.