Morbidity and mortality due to the rise in the incidence of fungal infections has increased significantly. The genus Candida is the most common cause of fungal infection worldwide. Recent evidence suggests that majority of infections produced by this pathogen are associated with biofilm growth. Biofilms are colonies of microbial cells encased in a self produced organic polymeric matrix. The biofilm production is more important for Non Albicans Candida (NAC); as C. albicans possess many other mechanisms to establish infections. Correct identification of Candida species has gained importance due to persistent rise in infections caused by NAC. The development of resistance in Candida is also raising due to the increase in immunocompromised status and critical patients. To study the prevalence of Candida species, biofilm formation for Candida isolates and their antifungal susceptibility pattern in a tertiary care centre.Sixty eight clinical isolates of Candida were collected from various clinical samples in a tertiary care centre, Chennai for a period of six months. Identification of Candida species was done by germ tube tests, test for chlamydoconidia production, sugar assimilation tests and CHROM agar isolation method. Antifungal susceptibility tests against fluconazole, Amphotericin B and Flucytosine were performed by disc diffusion method and for the resistant Candida strains Minimum inhibitory concentration (MIC) was performed using E-strip for fluconazole, Amphotericin B and Flucytosine. Out of 68 isolates C.albicans tops the first 67.6%, C. tropicalis ranks the next most predominant species 17.64% and the least common isolates C.parapsilosis 8.8% and C.glabrata 5.8%. Candidial infections were more commonly observed in female patients 57.3% and hospitalized in- patients 69.1% with increased number from non – catheterized urine samples. The Fluconazole susceptibility using disc diffusion showed 69% of C.albicans and 58.3% of C.tropicalis and 75.5% of C.parapsilosis were sensitive to fluconazole and sensitivity to Flucytosine were as follows C. albicans (86.60%) C. tropicalis (75%) C.parapsilosis and C.glabrata (100%).Though a higher degree of antifungal resistance noted among the various Candida species, still can be used as a treatment drug for critical care patients as the MIC of those anti fungal drugs were found to be in the susceptible range. Isolation of NAC from clinical samples, their biofilm formation and antifungal resistance are on a rise, they should be treated as a pathogen and not dismissed.