ABSTRACTThere are growing concerns that the use of performance-enhancing substances among amateur athletes may be on the rise globally. While most studies done in Nigeria have been limited to elite athletes, the fact remains that Nigeria university athletes constitute about 40% of the country’s representatives in international competitions. This study assessed the perception of university athletes about performance-enhancing substances and drug testing.A cross-sectional design was employed with the aid of semi-structured, self-administered questionnaires to gather data from 400 university athletes; information on socio-demographics, awareness and knowledge of performance-enhancing substances, perceptions about the use of performance-enhancing substances, drug testing and perceived factors influencing the use of performance-enhancing substances were obtained.The age range of respondents was 15–33 years with the majority (81.8%) of them being Christians. The majority (71.3%) of the respondents were aware of performance-enhancing substances; males (74.0%) and females (66.0%). About three-quarters (74.0%) had at least a fair knowledge of performance-enhancing substances. Two-thirds (68.0%) of the respondents disagreed that all athletes use performance-enhancing substances in competitions, while almost all respondents (90.5%) agreed that athletes can perform excellently at competitions without the use of performance-enhancing substances. A majority (81.3%) of the respondents agreed that there should be a ban on all performance-enhancing substances. Three-quarters (76.8%) identified fear and anxiety as the major factors influencing the use of performance-enhancing substances, while the majority were in support of and willing to undertake drug testing. Most Nigeria university athletes are aware of side effects and perceive drug testing as appropriate and relevant.