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This study was conducted to identify the prevalence of and potential risk factors for HIV-1 infection among drug users visiting drug treatment centers in Tehran, Iran.Cross-sectional quantitative study.Between October 2003 and May 2004, drug users who visited three public drug treatment centers in Tehran were interviewed and information about their sociodemographics, drug use characteristics, incarceration history, sexual history, and HIV/AIDS knowledge and risk perception were collected. Specimens of oral mucosal transudate were then collected from participants to be tested for HIV-1 antibodies. Logistic regression analysis was conducted on the association between the demographic and behavioral factors with HIV-1 infection.Overall, 611 (588 male and 23 female) drug users participated in the study. Among male injectors with HIV-1 prevalence of 15.2%, a history of shared injection inside prison [adjusted odds ratio (OR), 12.37; 95% confidence interval (CI), 2.94–51.97] was the main factor associated with HIV-1 infection. Among those who reported no history of injecting drug use, HIV-1 prevalence was 5.4%, and lack of condom use during sex was significantly associated with the infection (adjusted OR, 3.42; 95% CI, 1.25–9.36).HIV-1 infection is already prevalent among drug users in Tehran, Iran and shared injection inside prison has been revealed to be a particular risk factor for HIV-1 infection among injecting drug users. Harm reduction programs which have been started in Iran should be urgently expanded particularly in correctional settings and strengthened by condom use promotion to prevent sexual acquisition or transmission of HIV-1 among drug users.