Isolation and molecular characterization of Clostridium difficile strains from patients and the hospital environment in Belarus

Toxigenic Clostridium difficile is the most common etiologic agent of hospi tal-acquired diarrhea in developed countries. The role of this pathogen in nosocomial diarrhea in Eastern Europe has not been clearly established. The goal of this study was to determine the prevalence of C. difficile in pati ents and the hospital environment in Belarus and to characterize these isol ates as to the presence of toxin genes and their molecular type. C. diffici le was isolated from 9 of 509 (1.8%) patients analyzed and recovered from 2 8 of 1,300 (2.1%) environmental sites cultured. A multiplex PCR assay was u sed to analyze the pathogenicity locus (PaLoc) of all isolates, and strain identity was determined by an arbitrarily primed PCR (AP-PCR), The targeted sequences for all the genes in the PaLoc were amplified in all C, difficil e strains examined. A predominantly homogenous group of strains was found a mong these isolates, with five major AP-PCR groups being identified. Eighty -three percent of environmental isolates were classified into two groups, w hile patient isolates grouped into three AP-PCR types, two of which were al so found in the hospital environment. Although no data on the role of C. di fficile infection or epidemiology of C. difficile associated diarrhea (CDAD ) in this country exist, the isolation of toxigenic C, difficile from the h ospital environment suggests that this pathogen may be responsible for case s of diarrhea of undiagnosed origin and validates our effort to further inv estigate the significance of CDAD in Eastern Europe.