Abstract Introduction: Candidiasis is the commonest fungal infection affecting mucosa, skin, nails, and internal organs in the human host. Resistance to antifungal drugs is a growing problem, thus it important to study the burden of fungal infections and best therapeutic options available to treat them in a geographical location. Objectives: The purpose of this study was to determine the prevalence of infections caused by Candida species and its antifungal susceptibility profile in a tertiary care teaching hospital of rural Gujarat, India. Materials and Methods: This is an observational retrospective study. Data was collected for a period of five years (from January 2016 to December 2020) using Laboratory Information System (LIS) of Central Diagnostic laboratory of a tertiary care teaching hospital in rural Gujarat. The antifungal tested were Amphotericin B, Clo-trimazole, Fluconazole, Flucytosine, Voriconazole, Caspeofungin and Micafungin with isolated fungal species in our study. Results: A total of 53,195 culture samples were received during the study period with culture positivity rate of 37.27% (n=19,829) including bacterial and fungal isolates. Out of 19,829 isolates, 544 (2.74%) belonged to Candida species. Majority of candida isolates i.e., 70.8% were from critical care units. The most common species isolated was Candida tropicalis (44.30%) followed by Candida albicans (36.02%), Candida glabrata (8.08%) and Candida parapsilosis (4.59 %). The drug susceptibility of Candida tropicalis to Amphotericin B, Fluconazole, Flucytosine, Voriconazole, Caspeofungin and Micafungin were 96.4%, 91%, 95.4%,95.3%, 97.4% and 98.2% respectively. Percentage susceptibility of Candida albicans, ranged from 85.7% to 99.4% for various antifungal agents. The susceptibility rates for Candida glabrata ranged from 34.7% to 100% for all antifungal agents tested. The susceptibility rates for Candida parapsilosis were 42.8% to 100% for various antifungal agents tested. Conclusion: Majority of the infections were found in ICUs. Candida tropicalis was the most frequent isolate in our study and was highly susceptible to Micafungin i.e., 98.2%. Maximum resistance was found in Candida glabrata with Caspeofungin drug i.e., 30%.