Antibiotic-associated Clostridium difficile (CD) diarrhoea is one of the common causes of healthcare-acquired infection. Cephalosporins, piperacillin-tazobactam and aminoglycosides are the common antibiotics which have the maximum chances of producing Clostridium difficile infection (CDI). Most Asian countries have easy accessibility to many of these antibiotics without prescription. Broad spectrum antibiotics have been indiscriminately used as empirical therapy over the last two decades which has resulted in an increased risk of C. difficile infection. In India, the prevalence of CDI is highly underestimated. This study aims to understand the prevalence, risk factors and comorbidities associated with CD diarrhoea in a tertiary care hospital from western India. 196 patients were included in the study who were diagnosed with antibiotic-associated diarrhoea (AAD) clinically. Stool samples collected were processed for anaerobic culture of C. difficile and immunochromatography test was done to detect C. difficile toxins A and B. The comorbidities associated as well as the use of antibiotics like cephalosporin or proton pump inhibitors were also noted for the patients with CDI. 32 samples yielded CD (16%), out of which toxin production was detected only in 16 isolates. The prevalence rate of CDI in our hospital was 5%. Most of the patients had history of chronic illnesses like diabetes mellitus, chronic kidney disease, ischemic heart disease, systemic hypertension, autoimmune diseases, or malignancy. Avoiding empirical therapy with antibiotics prone to cause AAD, Antimicrobial stewardship programme with proper infection control practices and epidemiological surveillance of CDI will help to reduce the burden of CDI in our country.