Guatemala has a concentrated HIV epidemic disproportionately affecting men who have sex with men (MSM) and transgender women. We recruited 205 self-identified MSM, bisexuals, transvestites, and transgender women in western Guatemala using long-chain peer referral, wherein "seed" participants were asked to invite as many as three acquaintances to participate in the study. Self-reported sexual or gender identity was MSM, 46%; bisexual, 28%; transvestite, 21%, and transgender woman, 5%. Median age of the participants was 23years, and 36% self-identified as being indigenous. Indigenous persons were more likely to self-identity as transvestite (32.9% vs 13.8%, P = 0.04), strongly perceive themselves at risk for HIV (87.7% vs 51.5%, P = 0.001), have had an HIV test in the last 12months and know the result (97.3% vs 85.4%, P = 0.008), and experience barriers to testing and treatment (86.3% vs 67.7%, P = 0.004). HIV prevention services for indigenous MSM should especially target transvestites and how to overcome stigmatization and barriers to care.