BACKGROUND: Oral hygiene plays a pivotal role in maintaining overall health, with toothbrushes being a fundamental tool in daily dental care. However, little attention has been given to the critical issue of toothbrush contamination and its implications for public health. This study aims to understand the knowledge, attitude, and practice regarding toothbrush contamination and disinfection among the public. MATERIALS AND METHODS: This was a cross-sectional questionnaire study consisting of 15 questions that cover points of knowledge, attitude, and practice towards contamination and disinfection of toothbrush reporting was developed in Google form format among 350 participants who gave consent to participate in this study. The collected data was analyzed using the SPSS version 26 software package. RESULTS: There was a statistically significant gap regarding toothbrush contamination, mode of storing the toothbrush, and various practices used for toothbrush disinfection among each participant. It was also seen that general awareness and practice of toothbrush disinfection was higher among female participants in the study, which was also a statistically significant result. CONCLUSION: Understanding these aspects is essential in promoting healthier oral hygiene practices and subsequently improving public health. This study underscores the need for targeted educational campaigns and emphasizes the role of individuals in safeguarding their oral well-being.