The focus was on HIV-related knowledge, attitudes and practices among health workers in the city of Gao. Objectives: The aim was to assess knowledge of HIV among health workers in the city of Gao, to describe the practices of health workers in the management of HIV, and to assess the HIV training needs of health workers in the city of Gao. Methodology: We conducted a prospective cross-sectional study of staff in the city of Gao. Results: We surveyed 104 health workers. The majority of the staff surveyed were women (54.8%) with a sex ratio (F/H) = 1.21. The majority were between 30 and 39 years of age. In our series, 62.1% of staff surveyed knew about ARV treatment. Biological monitoring was cited as a tool for measuring CD4 counts by 45.2%. Our study led us to conclude that it would be advisable to provide ongoing training to improve knowledge and reduce HIV-related misconduct among health workers in the town of Gao.