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Background: Clostridium difficile ( C. difficile ) is a spore-forming, Gram-positive rod that is known to be associated with antibiotic use. It is one of the leading causes of nosocomial diarrhea in the industrialized world and therefore warrants further study of its nature. It isn't clear if co-infection by other organisms can affect the outcome of C. difficile infection (CDI)., Methods: A single center retrospective study was done and it used inclusion criteria of 18 years of age and being tested positive for CDI on FilmArray® multiplex gastro-intestinal (GI) panel. Exclusion criteria were a GI panel performed on an outpatient basis, recurrent CDI, and the presence of end-stage renal disease, cirrhosis, or a non-GI infection. The stool sample for all patients were collected within 48 h of presentation to the hospital. There were 235 of 2576 GI panels selected for a retrospective chart review based on the above criteria. Among these 235 patients, 38 had a co-infection (CDI+ another GI infection = group A or cases) and the rest had only CDI (group B or controls). Group A was compared with group B for CDI severity, its response to treatment, recurrence, and length of the hospital stay, using 0.05 as the alpha criterion., Results: Most patients with CDI were female and above the age of 60 years. Co infection did not increase the severity of CDI based both on the American College of Gastroenterology criteria (p 0.16) as well as Infectious Disease Society of America criteria (p 0.77). Co infection group also didn't have significantly different CDI related treatment failure rate (p 0.23), or CDI recurrence rate (p 0.49). Co-infection was also not associated with lengthier hospital stay (p 0.41)., Conclusion: Our study suggests that co-infection doesn't affect the severity of CDI or can cause treatment failures. Additionally, there was no significant increase in hospital stay, or increase in CDI recurrence associated with co-infection. Therefore, if CDI is the leading clinical diagnosis and a patient is tested positive for co-infection in addition to CDI on FilmArray® multiplex GI panel, this co-infection shouldn't change the management for CDI. Limitations of this study (including retrospective nature of the study, small sample size, single site study, not including all microbiome and non-inclusion of race) should also be taken into account, while considering the applicability of the results of this study., Competing Interests: Competing interestsThe authors declare that they have no competing interests., (© The Author(s) 2020.)