Background: A positive relationship between a healthcare provider (HCP) and a patient is likely to build trust and improve care for People Living with HIV (PLWH). The study explored the individual and external factors influencing HCP behaviour towards PLWH and compares HCP in antiretroviral therapy clinics (AHCP) with general health care settings HCP (GHCP). Methods: This qualitative study used a semi-structured interview protocol to guide individual in-depth interviews among 33 HCPs. The interview protocol was informed by empirical literature and included topics such as perceptions and experiences working with PLWH, HIV-related training received, knowledge about HIV, emotions towards PLWH and support related to patient care. The interview data were thematically analysed. Results: Both AHCP and GHCP claimed that their thoughts and judgments never influenced the way they managed PLWH, but there appears to be reflections of either sympathy or positive discrimination, especially for AHCP, or stigmatizing behaviours such as using gloves for PLWH-only among GHCP. The findings from this study suggest that individual factors such as perceptions about HIV, attitude towards HIV patients, different emotions, HIV-related training received and external factors such as availability of guidelines, logistics, infrastructural and reimbursement challenges influenced HCP behaviour towards PLWH. Conclusion: The study suggests that individual and external factors influence AHCP and GHCP behaviour towards PLWH. We recommend the use of Intervention Mapping to develop and evaluate interventions addressing the behaviour and emotions of AHCPs and GHCPs to reduce stigmatization of PLWH in the healthcare sector, hence improving hospital visits and medication adherence.