Purpose Candida albicans has been regarded as the most predominant oral fungal pathogen and the main cause of denture stomatitis. This study aimed to investigate C. albicans adherence to three types of denture base polymers: heat‐cured polymethylmethacrylate (PMMA), CAD‐CAM milled and 3D‐printed. The efficacy of four common disinfection techniques, glutaraldehyde, brushing, microwave irradiation, and Polident overnight tablets, were also examined. Material and methods Sixty blocks of pink acrylic specimens were fabricated from each polymer group. To investigate the C. albicans adherence, as well as the efficacy of different disinfection techniques on removing the yeast from the different materials, specimens were cultured within the fungal culture overnight followed by disinfection. The adhered C. albicans on the materials were then obtained by vortexing in phosphate buffered saline (PBS), and the numbers of the yeast in the suspensions were evaluated by measuring the optical density and/or colony‐forming units on agar plates. Data were expressed as mean ± SEM (standard error of the mean). Statistical differences were evaluated by one‐way analysis of variance (ANOVA) followed by the post hoc Tukey HSD tests. Results Significant differences in C. albicans adherence to the three polymers were noted. CAD‐CAM milled and heat‐cured PMMA showed significantly less C. albicans adherence compared with 3D printed PMMA. No significant difference was noted between milled and heat‐cured PMMA. In the disinfection test, microwave irradiation, mechanical brushing, and Polident tablets were found to be effective in removing fungal attachment on the different denture materials, while glutaraldehyde was found to be the least effective. Conclusion C. albicans adherence to the polymers varies greatly based on the types of PMMA. 3D‐printed had the highest fungal biofilm attachment. Microwave irradiation, mechanical brushing, and Polident overnight tablets had comparable results in removing C. albicans from all types of PMMA, while glutaraldehyde was not as effective.