Background: Egypt is reported to have the fastest growing epidemic of HIV in the Middle East and North Africa Region. Nurses who play a crucial role in delivering health care to persons with HIV/AIDS should be equipped with adequate knowledge about HIV/AIDS, in order to have sense of comfort to enable positive attitude towards such patients. Objective(s): The aim of this study was to assess knowledge and attitude regarding HIV/AIDS among nursing students in Sohag University. Methods: Across-sectional study was conducted among 324 randomly selected students of Faculty of Nursing, Sohag University, Egypt during the academic year 2019/2020. A self-administered questionnaire was used for collecting data about socio-demographic characteristics, HIV/AIDS knowledge and attitude of the study participants. Results: Most of the participants (78.4%)had a good overall knowledge about HIV/AIDS. However, some misconceptions were found as only 12.7% of them knew that HIV/AIDS cannot be prevented by vaccine. About 19% of the respondents mentioned that HIV cannot be transmitted by using toothbrushes. Less than one third were aware that HIV cannot be transmitted by either mosquito bites or by using swimming pools or toilets with infected persons and that persons may be symptoms free for more than 10 years. Concerning attitude, more than half of the students (52.6%) had negative attitude towards HIV/AIDS patients. On conducting logistic regression analysis, male sex and residing in urban areas were significant predictors of good knowledge and positive attitude towards HIV/AIDS, respectively. Information about HIV/AIDs was gained from TV (60.6%) followed by internet (45%), university curriculum (27%), friends (22%) and family (18%) Conclusion: The studied nursing students had moderately good general knowledge about HIV, but several misconceptions were prevalent among them. However, more than half of them exhibited negative attitude towards HIV/AIDS indicating urgent need for training programs to improve their knowledge and change their attitude towards persons with HIV/AIDS.