Background: Biosafety is a concern in all health sector services. Confronted with a high biological riskboth for patients and professionals in dentalcare and due to the constant development of new technologies, information, equipment, material and behavioral attitudes in this area. Aim: The aim of this study was to assess sterilization and infection control practices that are followed by private dental practitioners in East Delhi region. Methodology: A descriptive, cross-sectional survey was conducted to assess the attitude towards and sterilization practices among the private dental practitioners in East Delhi, India. A structured proforma was used for the collection of the information by the investigator. Results: A large proportion (68.0%) of the private dental practitioners carried out the pre-cleaning of the instruments before sterilization. Autoclave (66.0%) was the principle method of sterilization of the instruments among the private dental practitioners followed by Chemiclave (18.0%). Most (46.0%) of the private dental practitioners utilized the sterilized instruments within 1 week of sterilization. Autoclave was used more by the dental practitioners with more than 10 years of experience (35.7%) for sterilization of burs. Most (40.0%) of the private dental practitioners were not sterilizing endodontic files in their office. Most (52.0% and 66.0% respectively) of the private dental practitioners sterilized paper points and guttapercha respectively in their office. Conclusion: To achieve sterilization, it is necessary to adhere strictly to procedures for instrument cleaning and wrapping and to follow carefully the sterilizer manufacturer's operating and maintenance instructions.