Background: Stress in medical education has been inevitable among medical students. The prevalence of mental health problems among medical students is increasing, requiring a reliable tool for accessing assessments. The General Health Questionnaire (GHQ-12), designed to screen for mental disorders and psychological stress, is a promising tool. Although the factor structure of GHQ-12 has been studied internationally, it is still unclear in Vietnam. Objectives: To evaluate the reliability, validity, and factor structure of the Vietnamese version of the GHQ-12 questionnaire. Materials and method: An analytic cross-sectional study of the online Microsoft survey on 1763 students majoring in medicine, traditional medicine, and preventive medicine at Can Tho University of Medicine and Pharmacy from May 2022 to May 2023. Results: Samples in the study comprised 1763 respondents, out of whom male respondents were 42.8%, and female respondents were 57.2%. The average age was 22.62 with the highest proportion being the age group 18-29 years old (95.7%). Our research subjects were mainly studying medicine (86.5%), and the remaining disciplines included Traditional Medicine (8.1%) and Preventive Medicine (5.4%). The average GHQ-12 score was 16.22 points with the mean score obtained on item 5 (1.74) being the highest, indicating that most students were under strain. Cronbach's alpha coefficient of 0.893 showed an appropriate level of confidence. A two-factor structure with nine categories of “negative items” (2, 5, 6, 9, 10) and “positive items” (1, 3, 4, 8) had a cumulative variance of 60.175% drawn from the study. Conclusions: The Vietnamese version of GHQ-12 has enough reliability and validity to assess the mental health of medical students. The results indicated that students had higher-than-average scores, meaning poorer mental health. Furthermore, the study revealed a two-factor structure with nine categories including “negative items” and “positive items” with a cumulative variance of 60.175%. The two-factor structure suggests that GHQ-12 can be a multidimensional measure.