ABSTRACT The Africa Centres for Disease Control and Prevention Biosafety and Biosecurity Initiative being implemented across the 55 African Union Member States is presented. Based on consultations with Member States between 2019 and 2021, national-level capacity gaps were identified which informed the biosafety and biosecurity 5-year (2021–2025) strategic plan. The process of identifying national gaps, development, implementation and monitoring of the 5-year strategic plan is described. Notable achievements include development of a regional biosafety and biosecurity legislative framework now approved by African Union structures and process of domestication that has started in some countries; establishment and operationalisation of multi-sectoral regional biosafety and biosecurity technical working groups tasked with coordinating and monitoring implementation of the initiative, development and implementation of an accessible and regionally endorsed Regional Training and Certification Program for Biosafety and Biosecurity Professionals; establishment of a Regional Centre of Excellence for biosafety and biosecurity from where two cohorts of students in the two areas of biorisk management (16 students from 8 countries) and biological waste management (19 students from 10 countries) have been trained and development of a Regulatory and Certification Framework for Institutions Handling High-Risk Pathogens with three components of minimum standards of biosafety and biosecurity for high-containment facilities, a standard evaluation checklist for checking compliance and a certification framework for authorising performance of tasks related to dangerous pathogens. A guidance document with step-by-step process on how to translate regional successes to national implementation was developed and published. Based on notable regional successes of the initiative, Africa CDC co-chaired the Global Health Security Agenda Action Package 3 on Prevention and the Africa Signature Initiative Biosafety and Biosecurity Working Group. Challenges of delayed implementation due to the COVID-19 pandemic, limited resources to implement all planned activities and limited staff dedicated to biosafety and biosecuruty at Africa CDC.