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Background HIV/AIDS remains a modern-day global burden, killing almost 39 million people with over 36 million living with the disease, and two million new cases annually. The risk of infection in clinical practice among health workers has been noted to be high, compared to the general population. The study therefore seeks to determine the knowledge, attitude, and practice of HIV/AIDS prevention among nursing students along the north-eastern corridor of Ghana. Methods A cross-sectional descriptive design with a quantitative approach was employed. Multistage stratified random sampling technique was used to select 373 trainee nurses. A Pearson’s chi-square analysis was performed at 0.20 precision level. Determinants of knowledge, attitude, and practice was established with a multivariate logistic regression at 95% confidence level. Results Majority of participants had satisfactory knowledge (88.0%), good attitude (87.9%), and good practices (57.6%) towards HIV/AIDS prevention. Students in their final year of the study were 2.3 times more likely to demonstrate good attitudes towards HIV/AIDS prevention than first years (aOR: 2.28; CI: 1.10–4.72). Those with sufficient knowledge on HIV/AIDS were 3.5 times more likely to have good attitudes towards its prevention than those with insufficient knowledge levels (aOR: 3.46; 1.76–6.77). Conclusion There are satisfactory knowledge, attitude, and practices among trainee nurses regarding HIV/AIDS prevention in the study setting. Increase access to knowledge regarding HIV/AIDS can improve attitudes towards its prevention. However, there is a need for appraisal of the clinical placement programs of trainee nurses to ensure improved practice. Further studies are needed to ascertain the role of knowledge and attitudes in enhancing practices on HIV/AIDS prevention.