The results of the bio-behavioral monitoring study among prisoners were presented. The performed work is unique in its volume and in quality of the data obtained. Unlike all previous surveys in the institutions of the State Penitentiary Service, for the first time in Ukraine at the national level a coherent study was conducted on awareness and behavior of convicts as a component of the second generation HIV surveillance. According to a specially developed technique convicted were surveyed and tested for HIV-infection allowing the analysis of relationship between knowledge, behavior and the level of HIV-infection among detainees in penal institutions. National indicators for monitoring and evaluation of the effectiveness of HIV/AIDS control measures in penal institutions were also calculated, and recommendations were made to improve the strategy and tactics of HIV/AIDS prevalence prevention in prison facilities.