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To determine the prevalence and distribution of HIV in South African workplaces.Cross-sectional HIV prevalence and knowledge, attitudes and practices surveys, conducted in 22 public and private sector organizations in all nine provinces of South Africa on full-time, formally employed personnel who provided consent to participate.The primary outcome was HIV prevalence.The crude HIV prevalence among the 32 015 participants was 10.9%. HIV prevalence was higher among men (11.3%) than among women (9.8%) and among black Africans (16.6%) than among other race groups (2.7%). Although managers and employees with post-school education had a lower HIV prevalence than lower skilled employees, this only partly accounted for the race differences.The HIV prevalence within an organization is not entirely explained by the race, age and sex structure of the workforce. This indicates that there is some other factor that is associated with the organization and has an impact on HIV prevalence.