Clostridium difficile (Cd) infections rates and severity have been related to the emergence and spread of a hypervirulent toxigenic strain B1/NAP1/027. This strain has been related to outbreaks in North America and Europe, and most recently in Latin America. Our objective was to describe the prevalence of Cd strain BI/NAP1/027 among patients in Terciary care hospital in Cali-Colombia. Between January 2012 and May 2016 patients with Hospital acquiere diarrhea were screen for Cd. Molecular diagnosis of Cd strains was done through real-time polymerase chain reaction by detecting the toxin B gen, binary toxin gen and tcdC 117 nucleotide (GeneXpert; Cepheid, Sunnyvale, CA, USA). A total of 337 adult patients were studied. Median age was 61 years (25–87). Women accounted for 57.2%. The diagnosis of C. difficile was done in 48 patients (12.2%), of them 14 tested positive for the BI/NAP1/027 strain (29.2%). 92% of the cases scored severely. Two patients died due to Cd 027 strain infection (14.3%). Although the proportion of patients with C. difficile was low, almost one-third of them tested positive for BI/NAP1/027 strain. Emerging prevalence of Hypervirulent strains could be a major threat in Latin America, All authors: No reported disclosures.