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Long-term adherence to antiretrovirals is critical for sustained virologic response to HIV therapy in blood. Although antiretroviral therapy (ART) reduces HIV seminal shedding, little is known about the relationship between adherence to ART and HIV suppression in semen.To determine predictors of seminal HIV RNA suppression after 6 months of ART.Prospective observational cohort of 93 HIV-infected subjects before and after introduction of ART. Seminal HIV RNA was measured at baseline and 1, 2, 3, and 6 months after the introduction of therapy. Adherence to therapy was measured by self-report.A large academic HIV reference center in Rio de Janeiro, Brazil.Detectable HIV RNA in semen.In a multivariate logistic model with undetectable seminal HIV RNA after 6 months of therapy as the outcome variable, adjusting for baseline seminal viral load, both being adherent to therapy (OR = 11.8, P < 0.01) and using triple-drug ART (OR = 6.48, P = 0.04) were independently associated with seminal HIV RNA suppression.Inability to adhere to therapy was strongly associated with persistent shedding of HIV RNA in semen. Measures to improve adherence are urgently needed to reduce the sexual spread of potentially drug-resistant HIV among subjects using antiretrovirals.