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Introduction After the Taliban took over the Afghan government in August 2021, funds for the public healthcare system were suspended, the economy collapsed and female education and mobility were restricted. We examined the impact of the takeover on the availability, accessibility, acceptability and quality of public healthcare in Afghanistan.Methods We conducted a mixed-methods study to assess the impact of the Taliban takeover on Afghanistan’s public healthcare system. Key informant interviews with healthcare workers and managers were combined with interrupted time series analysis on maternal and child health services and the availability of medicines and healthcare workers from January 2019 to September 2023.Results Disruptions in service provision and availability of healthcare workers were minimal, temporary and mostly due to interrupted funding. Following the political change, more patients visited public healthcare facilities as safety improved accessibility and private healthcare became less affordable. Acceptability and quality of care were affected by continued increased patient load, medicine shortages and insufficient training planning. Healthcare workers reported incidents where chaperones obstructed access to healthcare facilities for female patients and healthcare workers.Conclusion Afghanistan’s public healthcare system showed incredible resilience amidst funding disruptions and conflict until 2023. Healthcare facilities and underserved communities are more accessible due to improved safety, but the increased patient load and economic collapse add pressure to the system. Development partners must safeguard access to and participation in healthcare for women.