This article presents the first national estimates of the overall burden of healthcare-associated infections and antibiotic resistance in NHS hospitals in Greece. It shows that, on any given day, about 9 out of every 100 patients hospitalized in Greece suffer from an infection acquired within the hospital. The total number of patients having a healthcare-associated infection on any given day in Greece was estimated at about 2,300 patients. This corresponds to approximately 121,000 newly affected patients each year in Greece. It was found that certain types of infections are associated with increased mortality, especially those caused by hard-to-treat superbugs (Carbapenem-resistant gram-negative bacteria) which double the risk of death in the hospital. Healthcare-associated infections were also shown to increase the length of hospitalization for more than 4 additional days on average, but for infections caused by carbapenem-resistant gram negative bacteria the increase exceeded 20 additional days of hospital stay.