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Toxinotyping of C. difficile strains 1234567 B1 PCR fragment 12349556778 A3 PCR fragment PCR method for screening changes in PaLoc markers for toxinotyping B1 B2 B3 PL3 A1 A2 A3 PL4 tcdRtcdBtcdAtcdEtcdC 2 kb 2.5 kb3 kb2 kb3 kb3 kb 2.1 kb2 kb1.5 kb2 kb1.6 kb H N HcHcHH R H R Ec P X hhS E E PL2 PL1 M. Rupnik Institute of Public Health Maribor Medical Faculty, University of Maribor© www.clostridia.net
Types of A-B+ C. difficile strains
C. difficile typing – mostly used methods Ribotyping REA PFGE (Europe) (USA) (North America) PCR of 16S-23S rDNA intergenic spacer region 160 ribotypes Stubbs, JCM 1999 Bidet, JCM 2000 HindIII restriction of whole DNA >100 REA groups (Rea Types) Gerding D., Chicago, USA SmaI restriction of whole DNA no large international collection © www.clostridia.net
Large clostridial toxins (LCT) C. difficile (TcdA, TcdB) C. sordellii (TcsH, TcsL) C. novyi (Tcn ) size (250-300 kDa)size (250-300 kDa) cytotoxicitycytotoxicity glycosyltransferasesglycosyltransferases autocatalytic proteolysisautocatalytic proteolysis © www.clostridia.net
C. difficile pathogenicity locus (PaLoc) © www.clostridia.net
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