Background: Clostridium difficile (C. difficile) is the most common antibiotic-associated diarrhoea agent in selected patients. C. difficile strains are classified as toxigenic or nontoxigenic based on their ability to produce toxins. Aim: The frequency of C. difficile and CDAD among patients in a tertiary hospital in Patna, India was investigated in this study. Methods: From the patients, a total of 233 diarrheal samples were extracted. The samples were cultivated on Clostridium difficile medium with cycloserine (500 mg/L), cefoxitin (16 mg/L), and lysozyme (5mg/L) and 5 per cent defibrinated sheep blood. Polymerase chain reaction (PCR) of the 16s rRNA gene identified the isolates as C. difficile, as did the presence of toxins genes (tcdA, tcdB, cdtA, and cdtB). The toxin production of isolates was then assessed using Rapid Card (CerTest BioTech SL Spain).