OBJECTIVE: This study aimed to assess the knowledge, attitude, and perceived barriers towards Evidence-Based Medicine (EBM) among healthcare professionals in southern India. METHODS: The study included doctors, dentists, nurses, and pharmacists who were willing to give informed consent. A total of 219 healthcare professionals were involved in the study. A questionnaire was prepared, and demographic details alongside the knowledge, attitude, and perceived barrier of the professionals towards EBM practice were collected and analyzed. The responses were scored, and the means were determined for each profession. RESULTS: The study found that dentists had the best knowledge about EBM practice, followed by doctors, pharmacists, and nurses. Pharmacists showed the best positive attitude towards EBM practice, followed by dentists, doctors, and nurses. Nurses were found to have the highest level of barriers to the practice of EBM, followed by doctors and pharmacists, while dentists tended to have the least barrier. CONCLUSION: The study recommends the need for increased awareness of the value and importance of EBM practice among healthcare professionals in southern India, and the provision of necessary requirements such as internet, computers, time, and training to increase the practice of EBM.