Introduction: Prevention of infection and control is an important part of safe patient care. Health-care professionals are at an increased risk of cross infection and its transmission while treating the patients. AIM: To assess the knowledge, attitude, practices regarding sterilization among health care staff in a dental college in Thodupuzha. Materials and Methods: Cross sectional study was done among 40 nursing and paramedical staff of Al Azhar Dental College, Thodupuzha. The study was carried out with the help of the questionnaire consisting 15 questions. Descriptive statistics was used for analysis. Results: Among the participants, majority of them reported changing lab coats if visibly contaminated and also of sterilizing instruments after each dental procedure and used protective barriers when assisting treatment procedure and only 55% of the participants admit to have had any basic training. Conclusion: The present study concluded that the knowledge, attitude and practice of nursing staff was not sufficient.