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The aim of the study was to determine the recurrence rate of denture stomatitis and persistence of Candida in 22 patients (5 male and 17 female, mean age 71 years) over a 3-year period.Denture hygiene practice, denture cleanliness, and the presence of palatal erythema were assessed for each patient at the start of the study (baseline). The oral cavity was sampled for yeasts by imprint culture and denture discs. Ten patients received a capsular form of itraconazole (100 mg twice daily for 15 days) and 12 patients were provided with 100 mg of itraconazole in the form of a mouthwash (10 mL twice daily), which was then swallowed. No further antifungal treatment was administered to any of the patients. Clinical and microbiological assessments were repeated for each patient at 6 months and 3 years after the original appointment. Yeasts were identified by colony color on CHROMagar Candida, germ-tube formation, and API-32C profiling. Selected isolates were then typed by inter-repeat polymerase chain reaction (IR PCR).Candida albicans was isolated at baseline from all patients either alone (12 patients) or in combination with another species (10 patients). Other yeast species recovered were C glabrata (5 patients), C tropicalis (1 patient), C guilliermondii (1 patient), C krusei (1 patient), C parapsilosis (1 patient), C kefyr (1 patient), and Saccharomyces cerevisiae (2 patients). Candida albicans and/or C glabrata were recovered from 11 of the 22 patients after 6 months or 3 years. A complete and consistent change of yeast species from baseline was observed in 6 patients after 6 months and at 3 years. The remaining 5 patients were yeast-free at the follow-up assessments. PCR fingerprinting of C albicans and C glabrata indicated strain persistence over 6 months in 10 patients and in 4 patients after 3 years. A switch in strain type occurred for 1 patient after 6 months and for 3 patients after 3 years.The recurrence of denture stomatitis in patients who maintained a high standard of denture cleanliness was low. Although itraconazole was beneficial in reducing the fungal load, there may be strain persistence or subsequent recolonization of the oral cavity by a broader range of potentially less sensitive yeast species.