The use of PCR ribotyping for molecular typing of clinically significant Clostridium difficile Romanian isolates

摘要

Clostridium difficile infections (CDI) became a significant infectious pathology in Romania in2010-2011, when first major outbreaks were reported. The increase of case numbers made moleculartyping of C. difficile strains necessary for epidemiological purposes. In this study, 96 C. difficile strains,isolated from CDI patients, were identified using PCR ribotyping. The method was selected consideringthe context of efforts being made to standardise C. difficile typing in Europe, using PCR ribotyping asthe method of choice, with a type nomenclature for cross-country comparison in CDI surveillance, butalso taking into account technical accessibility. The study results showed a majority of isolates (n=63)with the toxigenic profile A+/B+/CDT+, most of them (n=61) identified as hyper virulent ribotype 027.Eighteen more ribotypes were identified, in lower proportions. Fourteen corresponded to ribotypesfrom the Brazier nomenclature: 002, 012, 014, 015, 017, 018, 031, 046, 053, 085, 087, 106, 115 and140. Ribotype 014 was the most representative (7 isolates), followed by 002 (4 isolates). Three isolateswere non-toxigenic: ribotypes 031, 085 and 140. Other four ribotypes remained unidentified and werenamed U.01, U.02, U.03 and U.04. Two isolates clustering as ribotype U.02 were A+/B+/CDT+. Byselecting this typing method, a prevalent, highly epidemic ribotype and 18 other ribotypes weredetected, providing information about the C. difficile type diversity of the analysed group of strains andproving its usefulness as an instrument in epidemiological studies.