Objective To describe trends in AIDS-related knowledge and the prevalence of sexual behaviors among senior university students in Guangzhou city and to explore key issues in relevant publicity, education and intervention programs.

Methods We conducted a self-administered anonymous questionnaire survey among 6 202 senior students recruited with stratified multistage cluster sampling in 10 universities in Guangzhou city of Guangdong province between 2014 – 2018.

Results During the 5-year period among the students, the awareness rates of both overall and domain AIDS-related knowledge increased year by year significantly (P < 0.01) but 16.92% of the students reported heterosexual behavior, with an annual upward trend (χ² = 78.961, P < 0.001) . Of the male students with sexual behaviors during the period, 3.3% reported having homosexual anal sex in the previous one year. Condom use in every sex was reported by 58.1% (272/468), 62% (137/221), and 56.4% (22/39) of the male students having sex with regular, occasional, and commercial sexual partner during previous one year. The proportion of the students receiving and participating in AIDS publicity service increased yearly during the period significantly (χ² = 324.194, 32.383; P < 0.001). Multivariate logistic regression analysis revealed that main influencing factors for awareness rate of AIDS-related knowledge were whether having been involved in AIDS publicity service during previous one year (yes vs. no: odds ratio OR = 2.062, 95% confidence interval 95% CI: 1.621 –2.624) and age ( < 24 vs. ≥ 24 years: OR = 2.045, 95% CI: 1.607 – 2.602).

Conclusion Senior university students in Guangzhou city are lack of comprehensive AIDS-related knowledge and at a risk of having unsafe sexual behavior; the situation could be improved by promoting the students to have active participation in AIDS control publicity services.