The South African antiretroviral treatment guidelines recommend the use of human immunodeficiency virus type 1 (HIV-1) load testing for patient monitoring and, in particular, to assist in switching to second-line treatment regimens. There are significant challenges to implementing HIV load testing on the scale that is required in South Africa. To put this in context, approximately 560,000 HIV-infected individuals are receiving antiretroviral therapy, and program recommendations include viral load testing twice per year. Currently, a 3-tiered laboratory infrastructure exists with tertiary facilities and, to some extent, secondary laboratories able to implement quantitative HIV nucleic acid testing. Challenges include high sample volumes, transportation logistics from remote sites, costs, phlebotomy in children, a national skills shortage, and sample throughput of technology platforms. Several approaches are thus being explored simultaneously: (1) the feasibility of establishing higher throughput and more automated central laboratories; (2) improvement of current sample collection, transportation, and storage techniques; (3) alternative viral load technologies, including flow-based marker screening approaches to reduce testing volumes, and (4) point-of-care viral load testing strategies for clinics. The development of appropriate solutions for each laboratory tier in South Africa will require close collaboration between researchers in the field and partners in industry.