Denture-related stomatitis caused by Candida spp. affects elderly individuals using partial/total prosthesis, provoking several discomforts including burning sensation and altered taste. Herein, we have studied 52 denture-wearing individuals (>60 years-old), attended at the dentistry clinic of UNIVALE, aiming to isolate Candida spp. directly from the stomatitis lesions and to evaluate their potential to produce virulence attributes. A low prevalence of denture-related stomatitis was reported in these patients (4/52; 7.7%). Candida albicans was isolated in the 4 selected patients, with the ability to form biofilm over a polystyrene surface and to produce aspartic protease, esterase and hemolysin. However, neither phospholipase nor caseinase activities were detected. Planktonic-growing yeasts were susceptible to amphotericin B and caspofungin, while the susceptibility to azoles (fluconazol, itraconazole and voriconazole) varied depending on either the isolate or antifungal. Relevantly, biofilm-forming C. albicans cells exhibited resistance to all studied antifungals. So, new effective drugs against resistant C. albicans isolates causing denture-related stomatitis are urgently required.