Clostridium difficile is considered to be the causative pathogen in 20-30% of antibiotic-associated diarrhoea, and the leading cause of nosocomial diarrhoea, related to mortality rate of 1-5%. C. difficile is a gram-positive, anaerobic, sporogenous bacillus present in the soil, water and intestinal tract of animals, healthy children and adults. Vegetative forms of C. difficile, capable of toxin production, are found in the host's intestine, and when they are excreted to a dry and cooler outdoor environment with adverse conditions for survival, they transform into spores highly resistant to high temperature, desiccation and alcohol disinfectants, which can survive in the environment for up to 5 months.The most common mode of C. difficile transmission in the hospital setting is via contaminated hands of healthcare personnel. Strict and proper implementation of bundles of prevention measures can significantly prevent the emergence of nosocomial C. difficile infections (CDIs). The most important measures for prevention of nosocomial CDI include hygienic hand washing and disinfection of objects and surfaces with sporicidal chlorine disinfectants, however, the use of personal protective equipment and the proper procedure with hospital laundry are of great importance. A continuous education of medical and non-medical staff, patients and their family members is needed, as well as providing necessary material prerequisites so that the risk for nosocomial CDI transmission is minimized.