BackgroundAmong Latin America’s concentrated HIV epidemics, little is known about men who have sex with men (MSM) in Colombia, the region’s third largest country. To date, surveillance studies have been limited to Bogota, while 80% of HIV cases and deaths originate from Colombia’s other cities and departments. The extent to which interventions should prioritize MSM outside of Bogota is unknown.MethodsWe recruited 2603 MSM using respondent-driven sampling from seven of Colombia’s largest cities. HIV prevalence was estimated by site from dried blood spot samples. Behavioral data were collected through face-to-face interviews and risk factors for HIV infection analyzed using weighted, multi-level logistical regression models accounting for recruitment patterns.ResultsAcross cities, HIV prevalence averaged 15%, varied from 6% to 24% and was highest in Cali, Bogota, and Barranquilla. In the past 12 months, 65% of MSM had ≥ 5 casual male partners and 23% had a female partner. Across partnerships (i.e., casual, stable, and commercial), the proportion of MSM engaging in unprotected sex was ≥ 52% with male partners and ≥ 66% with female partners. Self-reported history of STI (24%) and past-year illicit drug use (38%) were also common. In multivariate analysis, age ≥ 35 (adjusted odds ratio [AOR], 19.2) and 25–39 (AOR, 5.6) relative to ≤ 18–24 years, identifying as homosexual relative to heterosexual (AOR 0.1), meeting casual partners on the Internet (adjusted odds ratio [AOR], 3.1) and age of sexual debut of ≤ 13 years (AOR, 3.1) predicted HIV infection. HIV testing and prevention messaging reached just 24% of MSM in the past year.ConclusionsFindings support consistently elevated HIV burden among MSM throughout Colombia’s largest cities and a need for enhanced behavioral prevention and HIV testing, emphasizing men who use the Internet as well as physical venues to meet sex partners.