Objective: To study the knowledge, attitude and practice of antibiotic usage and resistance among undergraduate medical students at Gauhati Medical College and Hospital. Methods: It was a questionnaire-based, cross-sectional and observational study. A structured validated questionnaire was used as a study tool consisting of 22 questions. A Google form was then developed and circulated. 3rd Professional Part-I and Part-II MBBS students were included in the study. Results: A total of 268 students participated in the study. Data were analysed using a Microsoft Excel Sheet and Student’s t-test. The majority gave correct responses regarding the meaning of antibiotic resistance and agreed that improper use of antibiotics leads to the development of antibiotic resistance. The majority (86.57%) feel that a combination of factors gives rise to antibiotic resistance. Overall, 259 (96.64%) of students think all healthcare providers should have a good knowledge of the rational use of antibiotics and that the use of antibiotics should be guided by culture and sensitivity reports. However, it was seen that only about 33% of students in both groups had attended an educational programme or training on the topic. Conclusion: Most students in both groups have good knowledge and attitudes regarding antibiotic usage and resistance. However, they must attend more educational programs and training to improve practice.