Background: Health care workers are key players in the prevention and management of HIV-infection. Materials and method: Descriptive cross-sectional hospital based study was conducted in Omdruman Teaching Hospital, Aim to assess the knowledge, attitude and practice of nurses towards HIV/AIDS’ patients, in the period between September to December 2019. Data was collected from 125 nurses, by using two methods: Self-administered questionnaire and observation check list. Results: Study result showed that, there was high knowledge about causative organism (96%), poor knowledge regarded the body fluids contains virus and methods of sterilization that kill the virus (19.2%). Satisfied knowledge was found about mode of transmissions, nursing procedures may lead to exposure to infection (72.8%), precaution measures taken by the nurses against HIV/AIDS while there was a poor practice of universal precautions. On the other hand, observation of hospital wards revealed insufficient measures to reduce the risk of HIV transmission (e.g. non-functioning water taps, lack of supplies and equipment). Study indicates there was a considerable percentage incidence of accidental needle sticks injuries (48.8%), but measures to be taken (post-exposure prophylaxis guidelines) after occurrence was poor. Conclusions: In general, negative attitudes and misconceptions were observed. 46.4% of nurses were afraid of caring an AIDS’ patients. It is suggested that continuous in-service training be instituted to improve practice, to dispel misconceptions and to develop favorable attitude.