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Objective: To evaluate the vulnerability of male prisoners to HIV, risk behaviour and access to prevention. Methods: This cross-sectional descriptive study was conducted in July and August 2012 in Ouagadougou Prison in Burkina Faso. Two trained investigators collected data by means of individual interviews in the prison visiting room using a questionnaire administered to male inmates 18 years and older, imprisoned for more than three months. Two focus groups were conducted with prison guards and healthcare personnel. Results: A total of 165 male prisoners were interviewed. The mean prison sentence was 19 months, the median age of the inmates was 28 years and 45% of them were illiterate. About 4% of male prisoners reported having had homosexual relations during their imprisonment. However, data indicate underreporting and denial of homosexual behaviour by prisoners. 49% of prisoners shared razors or razorblades in prison. None of the interviewees reported injected drug use or tattoos in prison. The majority (84%) of prisoners had a good knowledge of HIV/AIDS and 6% were aware of the risk of sexually transmitted infections. Only 5% of prisoners had had a screening test during their stay in prison. Conclusion: Prison conditions, homosexual behaviour and absence of condoms in prison accentuate the vulnerability of prisoners to HIV/AIDS. Implementation of a prevention programme and management HIV-positive prisoners would help to reduce significantly the risk of HIV transmission in prison.