ObjectiveTo understand the distribution of PCR ribotyps and toxin gene polymorphism of Clostridium difficile (C. difficile) in some areas of China, provide the reference sequences for the establishment of appropriate molecular detection and typing methods, and support clinical decision for the complex C.difficile infections.MethodsSixtyfour C.difficile isolates collected from 3 cities(Beijing,Guangzhou,Jinan) were performed PCRribotyping analysis. Among them, tcdA and tcdB of 26 representative strains were amplified and sequenced.ResultsAmong 64 C.difficile isolates, 45(70.31%) were type A+B+,19(29.69%) were type A-B+. Nine different PCR ribotypes were obtained. The top 3 ribotypes were 017(21 isolates,32.81%), 001(13 isolates,20.31%), and 012(11 isolates,17.19%). Among A-B+ strains, 14（73.68%）were ribotype 017, 1 was ribotype 001. Toxin gene sequences showed polymorphism, there were 7 TSTA, 6 TSTB, and 8 TSTG.ConclusionThe dominant Chinese ribotype may be 017, and toxin genes show certain polymorphism among strains, there is a corresponding relationship between PCR ribotypes and toxin gene polymorphism. Further study will be performed with expanding strains, so as to find appropriate molecular detection and molecular typing method, and help hospitals to prevent and control C. difficile infection.