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Health Care Workers (HCWs) face a grave risk of acquiring infection including Human Immunodeficiency Virus (HIV) and Hepatitis B & C during the course of their work. A questionnaire based study was conducted amongst the 154 HCWs working in a teaching hospital to study Acquired Immunodeficiency Syndrome (AIDS) awareness, and to assess the knowledge, attitude, and practice in respect of application of universal precautions. This study has shown that definite lacunae exist regarding awareness and practice of universal precautions. Comprehensive in service educational programme for HCWs regarding the implementation of universal precautions is recommended.