Clostridium difficile is responsible for more than 90 % of cases of antibiotic-associated diarrhea and pseudomembranous colitis. The most important virulence factors are two toxins called enterotoxin A and cytotoxin B; some C. difficile strains contain the C. difficile binary toxin (CDT). The aim of our study was to prospectively analyze C. difficile clinical isolates in a single center to determine the molecular features of collected strains. Among the 252 isolates, 217 were A + B + (86.1 %), 33 were A + B + cdt + (13.1 %) and 2 were A - B + (0.8 %). There were 15 different ribotypes with a predominance of 018.