Background: Clostridium difficile infection (CDI) is a common cause of diarrhea in hospitalized patients worldwide however it is uncommonly reported from Indian hospitals and clinical profile of Indian patients with CDI is not well studied. Methods: This was a retrospective study done in a tertiary care centre of South India between March 2016 and May 2017. Case records of all patients for whom stool samples were sent for either C. difficile toxin assay or GeneXpert C. difficile were analysed. Coloured immunoassay (CerTech) was used for detecting Glutamate dehydrogenase (GDH) and Toxin A and B and nucleic acid amplification assay (Xpert, Cepheid) were used. Results: A total of 112 were analyzed, out of which 18 were positive and negative cases (n = 94) were taken as controls. Prevalence of CDI was 16%. Presence of fever (OR = 8.5), tenesmus (OR = 8.5), hematochezia (OR = 4.5), cramps (OR = 8.27), use of immunosuppressive agents (OR = 3.62) and duration of antibiotics (7.38±6.50 vs 4.54±6.57 days; p=0.019) was significantly higher among cases. Conclusion: Although the prevalence of CDI is low in India compared to the west, it still contributes to significant morbidity in hospitalized patients as it is responsible for more than 1 in 6 cases of diarrhea in hospitalized patients. Fever, cramps, tenesmus, hematochezia, use of immunosuppressants and antibiotic therapy of more than 7 days suggest a diagnosis of CDI, and should trigger testing and treatment. Outcomes are good with appropriate therapy.