Objective: To assess the prevalence of HIV infection, to highlight HIV-testing refusal rates among participants in a population-based tuberculosis survey and to assess the implication for programme implementation.Methods: This cross-sectional study on the characteristics of participants who refused HIV testing was conducted in a national survey in Zambia. All eligible participants were aged above 15 years and included in the analysis.Results: Out of the 44 791 tuberculosis survey participants, 14 164(31.6%) refused to participate in HIV testing. The unemployed, rural dwellers, married, and those aged 15-24 years were associated with higher refusal rates.Conclusions: Strategies to improve HIV testing acceptance are necessary. Qualitative research is recommended to understand the reasons for testing refusals so that remedial interventions can be implemented.