Compared to HIV research on men who have sex with men, less is known about the risks and vulnerabilities for HIV among Male to Female (MTF) transgender persons, particularly in different geographic regions like Mexico. In Tijuana, Mexico, a border city experiencing a dynamic HIV epidemic, no precedent data exists on the MTF transgender population. Our aims were to estimate HIV prevalence and examine the behaviors and characteristics of the population. We conducted a cross-sectional study of 100 MTF transgender persons recruited through time location sampling in 2012. Participants underwent interviewer-administered (paper and pen) surveys and rapid tests for HIV. Descriptive univariate analyses were conducted on various factors, including sociodemographics, substance use, accessing social services (requested vs. received), stigma, and sex behaviors. A total of 22% tested positive for HIV, a prevalence higher than other key populations at risk for HIV in Tijuana.