Aims and objectives: Onychomycosis is a chronic fungal infection of nails. The clinical diagnosis of onychomycosis needs to be confirmed by direct microscopy and culture for identification of specific pathogen and proper selection of antifungal treatment. The study aimed to determine the prevalence of onychomycosis and cultural positivity of onychomycosis with respect to various etiological agents. Introduction: Dermatophytes are the most common pathogens of onychomycosis, yeasts and nondermatophyte moulds can also be found as causative agents of onychomycosis. Materials and methods: Nail clippings from 90 clinically suspected cases of onchomycosis were collected from OPD at Department of Dermatology, Venerology and Leprology, SMS and ESIC hospital, Jaipur. Detailed history of trauma, infection, occupation, diabetes, personal habits (smoking etc.) were taken Diagnosis was based on direct microscopy by 20% KOH examination method and fungal culture on SDA and Dermatophyte Test Media. Results: Out of 90 cases of onychomycosis patients, 80 (88.9%) were found KOH positive and 33 (36.7%) were positive in culture. In the present study, dermatophytes, nondermatophytes and yeasts were found etiological agents of onychomycosis. Distal subungual onchomycosis was found common type of onychomycosis in patients. Among the culture positive cases, Trichophyton rubrum was found the most prevalent species followed by T.mentagrophyte and T.tonsurans. Nondermatophyte molds were also isolated from white superficial onychomycosis and Candida species was isolated from Candida onychomycosis. Majority of patients were more prevalent in the age group of 41-50 and 51-60 years and males were commonly affected in our study. In the study, it was reported that, majority of patients that are infected with onychomycosis infection were service class and sportsmen followed by labours, housewives, unemployed, students and farmers. Conclusion: Based on this study it could be concluded that, onychomycosis is an important public health problem and accurate diagnosis of onychomycosis by direct microscopy and fungal culture is an essential requirement for appropriate management of such cases.