Vancomycin-resistant enterococci (VRE) have been a major public health concern for years. These pathogens are highly resistant, which facilitates their spread in health care settings where they may cause infections difficult to treat with antibiotics, particularly in immunocompromised patients. Over recent years, a considerable increase in VRE has also been observed in the Hradec Králové University Hospital. In 2017, 59 cases of VRE colonization or infection were newly diagnosed in inpatients. Based on a retrospective investigation, infected patients were identified, and from the hospital information system data, clinical and epidemiological factors possibly linked to VRE were analysed. It was concluded that one of the reasons behind the increase in VRE was poor compliance with the hygienic antiepidemic rules in general and with hand hygiene in particular, which was confirmed by the changed epidemiological situation the following year after more stringent rules were adopted. Raising awareness of health care workers about good practices to reduce the risk of the spread of resistant pathogens is crucial for the prevention of these infections.