Clostridium difficile Occurrence, Toxin Profile and Antibiotic Susceptibility: An Egyptian Center Experience Background: Clostridium difficile (C. difficile) infection (CDI) is of concern in healthcare facilities. The role of binary toxin alone is controversial. Resistance to antibiotic therapy has emerged. Objectives: to determine the occurrence of C. difficile, their toxigenicity, antibiotic susceptibility pattern and the effect of binary toxin gene. Methodology: A cross section study, stools samples were collected from 211 patients. Strains were identified, antibiogram was determined using ATBTM ANA, and E-test for vancomycin. A multiplex PCR assay for (tpi), tcdA and tcdB genes, followed by a duplex PCR for cdtA and cdtB genes were done. Results: C. difficile was isolated from 29 samples (13.7%). One strain was resistant to metronidazole, and three had intermediate susceptibility. All isolates were positive for (tpi) gene. Fifteen isolates (51.7%) were non toxigenic and 14 (48.2%) were toxigenic. Three toxigenic isolates harbored binary toxin genes. It was not possible to identify any impact of binary toxin due to small number found to harbor it. Conclusions: C. difficile is not a major problem in our locality. Further studies are needed to detect the role of binary toxin. Proper infection control measures and rationale use of antimicrobials are mandatory to prohibit further exaggeration of the problem.