Background: This study was conducted for a comparative evaluation of antimicrobial efficacy of commercially available four types of denture cleanser. Material and methods: In this study, the antimicrobial efficacy of three types of denture cleaning agents was assessed. The agents used were named Viclean (Group D1), Clinsodent (Group D2) and Fittydent (Group D3). The control group comprised of distilled water. A custom metal mold was created to produce wax plates with consistent dimensions of 15 mm × 15 mm and a thickness of 1.5 mm, which were subsequently transformed into heat-cured acrylic resin samples. The mold featured a square window measuring 15 mm × 15 mm and a thickness of 1.5 mm, designed to replicate the thickness of standard dentures.To facilitate the process, the metal mold was coated with a thin layer of petroleum jelly and positioned on a glass slab that had also been treated with petroleum jelly. Molten modeling wax was poured into the mold's window, and a second glass slab, similarly coated with petroleum jelly, was promptly placed on top of the metal mold. The molten wax was allowed to solidify without disturbance. Once fully hardened, the top glass slab was carefully slid off the metal mold, followed by the removal of the metal mold from the first glass slab, allowing for the extraction of the wax sample.Fungal cells adhering to acrylic resin surfaces of samples (Group D1) were fixed with formaldehyde. Samples along with yeast cells adhered to them were examined and the total number of colonies formed over sample was counted by microscopy. Now the samples were immersed in freshly prepared denture cleanser solution, D1 for 8 h. After 8 h, again samples were examined and total number of colonies remaining on sample was counted. The same procedure was performed using denture cleansers D2 and D3. For the control group, samples were immersed in distilled water for 8 h. Results: Control solution showed almost no change in number of count pre- and post-treatment. Denture cleanser solution D2 on the other hand showed highest reduction in the number ofcounts.Among three denture cleanser solution, D1and D2 showed a greater decrease in percent OD when compared with D3. Conclusion: Control solution showed almost no change in number of count pre- and post-treatment. Denture cleanser solution D2 on the other hand showed highest reduction in the number ofcounts.Among three denture cleanser solution, D1and D2 showed a greater decrease in percent OD when compared with D3.