Toxigenic Clostridium difficile culture is considered to be the standard diagnostic method for the detection of C. difficile infection (CDI). Culture methods are time-consuming and although enzyme immunoassay is rapid and easy to use, it has low sensitivity. In the present study, the AdvanSure CD real-time (RT)-PCR kit (LG Life Sciences) was evaluated for its ability to detect C. difficile toxin A (tcdA) and B (tcdB) genes, simultaneously. A total of 127 fresh diarrhoeal stool specimens, submitted to the clinical microbiology laboratory for C. difficile culture, were tested. C. difficile toxins and toxin genes were detected with a VIDAS C. difficile A&B (VIDAS-CDAB) enzyme-linked fluorescent immunoassay (ELFA) and the AdvanSure RT-PCR kit, respectively, according to the manufacturers' instructions. Their performance was compared with a standard toxigenic culture method as a reference. The sensitivity, specificity and positive and negative predictive values using the AdvanSure RT-PCR kit were 100 %, 98.3 %, 84.6 % and 100 %, respectively, while those of the VIDAS-CDAB system were 63.6 %, 100 %, 100 % and 96.6 %, respectively. Four tcdA(+)/tcdB(+) strains of C. difficile were detected with the AdvanSure RT-PCR kit, which offers comparable sensitivity and specificity to the reference method with a turnaround time of ~3 hours.