The health service sector has a vital role to play in delivering human immunodeficiency virus (HIV) and tuberculosis (TB) prevention, treatment and care, yet evidence indicates that healthcare workers (HCWs) themselves lack adequate access to HIV and TB services. HCWs are also at increased risk from TB and other infectious diseases at work, and therefore accessing HIV services is particularly important. A systematic review was therefore conducted to inform the development of World Health Organization (WHO) guidelines to improve access to HIV and TB services, and specifically, to assess the evidence regarding providing such services through workplace-based programmes. We identified any study published since 1984 that addressed outcomes of interest as defined through multi-stakeholder consultations, and were related to workplace interventions in (1) the healthcare workplace and (2) any workplace that included HIV and/or TB diagnosis and/or treatment. Interventions focusing solely on primary prevention with no diagnostic or treatment services were excluded, as they were the subject of other guidelines. A minimum of two reviewers independently extracted data and assessed the articles against pre-set selection criteria; studies were also profiled and quality assessed by a minimum of two reviewers. Three studies met these criteria specifically for HCWs; all showed a preponderance of positive benefits, with minimal negative outcome. Seven studies met these criteria regarding workplace HIV and/or TB diagnosis and/or treatment from other sectors, public or private. Again, all showed positive results. The paucity of high-quality evidence in this field of research was itself an important finding, beckoning further research on workplace-based programmes for health workers. Nonetheless, while more well-designed intervention studies are definitely desirable, providing programmes for HCWs to obtain HIV and TB diagnosis and treatment at the workplace is supported by the literature and is consistent with the values of the stakeholders, justifying the WHO-International Labour Organization-UNAIDS guidelines that emerged.