Background Medical students are considered to be personnel with a high level of risk for developing latent tuberculosis infection (LTBI). One possible reason is lack of knowledge about the transmission, prevention, and biosafety standards for tuberculosis disease. Objective This research aimed to determine the rate of LTBI among medical students studying in a private School of Medicine in Monterrey, Mexico. Methods In this cross-sectional study, we obtained blood samples from 174 medical students. LTBI was diagnosed using the QuantiFERON®-TB Gold Plus test. The prevalence of LTBI was compared with the socio-demographic data of the students and their level of knowledge and use of personal protective equipment (PPE). Results The proportion of LTBI in the students was 20.6%. Medical students in their first few years of medical school had a lower prevalence of LTBI than students in their final years of medical school. Additionally, students with a low level of knowledge on LTBI and low use of proper PPE had a higher prevalence of LTBI. Conclusions In a School of Medicine in Monterrey, Mexico, the proportion of medical students with LTBI was low but the proportion increased in advanced students. Students who demonstrated adequate knowledge and use of respiratory protective masks had lower prevalence rates for LTBI.