ABSTRACTThis study assessed the knowledge, acceptance, and uptake of Premarital Genotype Counselling and Testing (PGCT) among Youth Corps members in Osogbo, Nigeria. A descriptive cross-sectional design was employed. A validated, semi-structured questionnaire was used to assess knowledge, acceptance and uptake of premarital genotype counselling and testing. The sample consisted of 333 youth corps selected from the National Youth Corps Secretariat in Osogbo, Osun State. Descriptive and inferential statistics were used for data analysis. Most respondents (67.0%) had good knowledge of PGCT. Acceptance was high (71.8%). Uptake was notably high, with the majority (74.8%) reporting having undergone testing. Statistically, good knowledge was significantly associated with high acceptance (χ² = 21.587, p < .001), and high acceptance was significantly associated with increased uptake (χ² = 18.367, p < .001). No significant associations were found between sociodemographic variables (age, gender, marital status, religion) and uptake. Low partner discussion (39.3%) was perceived to be a critical barrier to testing. Knowledge, acceptance, and uptake of PGCT were high. However, communication gaps between partners were perceived as affecting testing uptake. Targeted education and couple-based counselling are needed to enhance understanding and maximise the benefits of PGCT among Nigerian Youths.