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ObjectiveTo assess the impact of an interprofessional case-based training programme to enhance clinical knowledge and confidence among clinicians working in high HIV-burden settings in sub-Saharan Africa (SSA).SettingHealth professions training institutions and their affiliated clinical training sites in 12 high HIV-burden countries in SSA.ParticipantsCohort comprising preservice and in-service learners, from diverse health professions, engaged in HIV service delivery.InterventionA standardised, interprofessional, case-based curriculum designed to enhance HIV clinical competency, implemented between October 2019 and April 2020.Main outcome measuresThe primary outcomes measured were knowledge and clinical confidence related to topics addressed in the curriculum. These outcomes were assessed using a standardised online assessment, completed before and after course completion. A secondary outcome was knowledge retention at least 6 months postintervention, measured using the same standardised assessment, 6 months after training completion. We also sought to determine what lessons could be learnt from this training programme to inform interprofessional training in other contexts.ResultsData from 3027 learners were collected: together nurses (n=1145, 37.9%) and physicians (n=902, 29.8%) constituted the majority of participants; 58.1% were preservice learners (n=1755) and 24.1% (n=727) had graduated from training within the prior year. Knowledge scores were significantly higher, postparticipation compared with preparticipation, across all content domains, regardless of training level and cadre (all p6 months, knowledge and self-reported confidence scores were greater compared with precourse scores (all pConclusionTo our knowledge, this is the largest interprofessional, multicountry training programme established to improve HIV knowledge and clinical confidence among healthcare professional workers in SSA. The findings are notable given the size and geographical reach and demonstration of sustained confidence and knowledge retention post course completion. The findings highlight the utility of interprofessional approaches to enhance clinical training in SSA.