These guidelines form part of efforts to institutionalize the prevention and containment of antimicrobial resistance (AMR) in healthcare facilities in South Africa, as outlined in the Antimicrobial Resistance Strategic Framework and Implementation Plan. The focus of these guidelines is on two interrelated aspects of prevention of healthcare associated infections (HAIs) and their spread; and the application of antimicrobial stewardship (AMS) practices at hospital level. They aim to serve as a practical, step-by-step or ‘how-to’ guide, addressing the infection prevention and AMS components of a robust response in a hospital. They draw on evidence from various international guidance documents and standards for interventions that have been shown to be successful in infection prevention and AMS programmes. These interventions have been customised to the South African hospital setting based on local experiences in the public and private health sectors. This was done through a series of workshops and requests for comment involving country-level experts.