Background and objective: Dermatophytes, fungi causing superficial skin diseases, cause millions of global infections, with a 10--20% lifetime risk of contracting fungus.The study aimed to identify and isolate dermatophytes from suspected cases of dermatophytosis, evaluate KOH microscopy and culture results, and diagnose dermatophyte infection.Material and method:A cross-sectional study involved 404 patients suspected of having dermatophytosis at Maharaja Yashwantrao Hospital in Indore, Madhya Pradesh. Samples were collected from December 2022 to January 2023, and the study was approved by scientific and ethical committees.Samples of skin, hair, and nails from 404 clinically suspected cases of dermatophytosis in both sexes and all age groups were collected, processed with KOH, and cultured on Sabouraud dextrose agar and Dermatophyte test media. Result: The study analyzeddermatophytosis in patients, with a slightly female preponderance. The most common clinical presentation was Tinea corporis (53.9%), followed by Tinea cruris (24.5%). Direct microscopy results showed 232 (57% of cases) were KOH positive, with a significant association between KOH mount and culture findings. The sensitivity of KOH positivity was 84.7%, and the specificity was 83.6%. The positive predictive value and negative predictive value were 87.14% and 80.70%, respectively.Conclusion: In conclusion, combining KOH microscopy with fungal culture in diagnostic strategies enhances the precision and speed of detecting dermatophytes in medical settings. Future studies could explore new diagnostic approaches or techniques for continuous improvement.