The article provides information on an outbreak of infection with a strain of Staphylococcus aureus resistant to gentamicin (SARG) and a hyperproducer of enterotoxins A and D at a neonatal intensive care unit of the University Hospital in Hradec Králové. A retrospective investigation was carried out that provided an overview of the presence of the SARG strain in the patients, staff and hospital environment. Subsequently, strict infection precautions were implemented and the importance of rational antimicrobial therapy was emphasized.