Aim: The purpose of this study was to evaluate 2% chlorhexidine disinfectant (CHX), chitosan, and octenidine dihydrochloride (as cavity disinfectants) on microleakage in cavities restored with universal self-etch adhesive. Materials and methods: Eighty extracted human permanent premolars were selected. Class V cavities were prepared on the facial surface of each tooth. The teeth were then divided into four groups of 20 teeth each. For the control group after cavity preparation, no disinfectant was applied. The other 3 groups were treated with 0.1% chitosan, 2% CHX, and 0.1% octenidine dihydrochloride (OCT). All the groups were restored with universal adhesive followed by composite resin. The teeth were then immersed in 1% methylene blue dye and were sectioned buccolingually. Microleakage was checked under a stereomicroscope on both occlusal and gingival margins. Result: Among all the groups chitosan-treated cavities showed the least microleakage. Chlorhexidine treated cavities showed less leakage as compared to control, OCT group at both the margins. Conclusion: Chitosan as a cavity disinfectant improves the sealing ability of the self-etch adhesive. Furthermore, in vivo studies need to be conducted to examine the interaction and long-term effect of chitosan with the other self-etch adhesive systems. Clinical significance: Chitosan a natural polysaccharide can be used as a cavity disinfectant as it improves the sealing ability of self-etch adhesive.