In this study, the rapid detection of Clostridium difficile and its toxins (A/B) in admitting patients in hospitals were offered the possibility of prevention of the bacterium spread into other patients. This bacterium is the most common cause of hospital-acquired diarrhea in patients treated with antibiotics, chemotherapeutic agents and other drugs that alter the normal equilibrium of the intestinal flora. The Stool samples were cultured on specific agar media with anaerobically growth equipment. The C. difficile toxins A and B were detected in fecal samples by using enzyme-linked immunosorbent assay (EIA). In this study, there was a higher incidence of C. Difficile infection among the hospitalized patients in different hospital wards, especially in the renal ward, hepatic ward and oncology ward. The using of EIA technique for pathogen detection is very useful and significant method for the rapid detection of the C. difficile strains.