Isolation and ribotyping of Clostridium difficile from patient stool samples

Clostridium difficile infections range from mild diarrhea to life-threatening pseudomembranous colitis. Recent outbreaks of Clostridium difficile infections are characterized by increased severity, high relapse rates, and significant mortality in North America and Europe. These infections have been related to the emergence of new hypervirulent C. difficile strain characterized as PCR – ribotype 027. More recently, another hypervirulent C. difficile strain PCR – ribotype 078 has emerged. The distribution of PCR ribotypes varies among European and North American countries. There has recently been ongoing European Clostridium difficile infection surveillance in the European Union, including Slovakia. The current situation on the presence of hypervirulent strains of C. difficile in Slovakia is unknown. Presently in hospitals, a patient's clinical symptoms and toxin positive diarrheal stool are used to diagnose C. difficile infection. This poses the problem of false cytotoxicity test results, and no isolation and further characterization of C. difficile isolates are done (e.g. ribotype, antimicrobial resistance profile). To begin collecting this data on the C. difficile ribotypes found in Slovakia, fifty samples of C. difficile toxin positive patient stools obtained from the HPL Microbiological Laboratory will be tested. If the stool sample is found to contain C. difficile, the bacteria will be isolated and the PCR ribotyping method for C. difficile will be introduced and recovered isolates of C. difficile will be characterized. The profile of these isolates from Slovakia will be compared to C. difficile isolates from other countries in the European Union as well as the United States and Canada.