Sexual prejudice toward men who have sex with men (MSM) is a pressing concern in sub-Saharan Africa (SSA). Given the high HIV infection risk among this population, sexual prejudice perpetuated by healthcare providers, affects access to and willingness of MSM to seek HIV care services. However, data on healthcare providers’ attitudes towards MSM in SSA are limited, and there are no locally-adapted measures of sexual prejudice. We adapted a scale to measure sexual prejudice with a sample of 147 healthcare providers in western Kenya. Results from exploratory factor analysis revealed a single-factor structure. The scale demonstrated high internal consistency with Cronbach’s α = 0.91. Healthcare providers who had prior interpersonal contact with MSM, had ever been trained on counselling MSM, and had higher knowledge about MSM health needs reported lower sexual prejudice scores, compared with peers who lacked these experiences (p