Abstract
Clostridium difficile shows considerable variability in the PaLoc region encoding two main virulence factors, toxins TcdA and TcdB. Strains with changes in PaLoc are defined as variant toxinotypes and currently 27 such groups are recognized (I to XXVII). Toxinotype 0 includes strains with PaLoc identical to the reference laboratory strain VPI 10463. Toxinotyping is a RFLP-PCR-based method using a combination of restriction patterns of part of tcdB and tcdA genes for determination of toxinotype. Variations in PaLoc can affect the toxin production or could result in production of toxins with altered properties.
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References
Braun V, Hundesberger T, Leukel P, Sauerborn M and von Eichel–Streiber C. (1996) Definition of the single integration site of the pathogenicity locus in Clostridium difficile. Gene 181, 29–38.
Rupnik M, Dupuy B, Fairweather NF, Gerding DN, Johnson S, Just I, Lyerly DM, Popoff MR, Rood JI, Sonenshein AL, Thelestam M, Wren BW, Wilkins TD and Eichel-Streiber CV. (2005) Revised nomenclature of Clostridium difficile toxins and associated genes. J Med Microbiol 54, 113–117.
Borriello SP, Wren BW, Hyde S, Seddon SV, Sibbons P, Krishna MM, Tabaqchali S, Manek S and Price AB. (1992) Molecular, immunological, and biological characterization of a toxin A-negative, toxin B-positive strain of Clostridium difficile. Infect Immun 60, 4192–4199.
Lyerly DM, Barroso LA, Wilkins TD, Depitre C and Corthier G. (1992) Characterization of a toxin A-negative, toxin B-positive strain of Clostridium difficile. Infect Immun 60, 4633–4639.
Depitre C, Delmée M, Avesani V, L’Haridon R, Roels A, Popoff M and Corthier G. (1993) Serogroup F strains of Clostridium difficile produce toxin B but not toxin A. J Med Microbiol 38, 434–441.
Rupnik M, Avesani V, Janc M, von Eichel–Streiber C and Delmée M. (1998) A novel toxinotyping scheme and correlation of toxinotypes with serogroups of Clostridium difficile isolates. J Clin Microbiol 36, 2240–2247.
Rupnik M. (2008) Heterogeneity of large clostridial toxins: importance of Clostridium difficile toxinotypes. FEMS Microbiol Rev 32, 541–555.
Spigaglia P and Mastrantonio P. (2002) Molecular analysis of the pathogenicity locus and polymorphism in the putative negative regulator of toxin production (TcdC) among Clostridium difficile clinical isolates. J Clin Microbiol 40, 3470–3475.
Curry SR, Marsh JW, Muto CA, O’Leary MM, Pasculle AW and Harrison LH. (2007) tcdC genotypes associated with severe TcdC truncation in an epidemic clone and other strains of Clostridium difficile. J Clin Microbiol 45, 215–221.
Chaves-Olarte E, Freer E, Parra A, Guzman-Verri C, Moreno E and Thelestam M. (2003) R-Ras glucosylation and transient RhoA activation determine the cytopathic effect produced by toxin B variants from toxin A-negative strains of Clostridium difficile. J Biol Chem 278, 7956–7963.
Perelle S, Gibert M, Bourlioux P, Corthier G and Popoff MR. (1997) Production of a complete binary toxin (actin-specific ADP-ribosyltransferase) by Clostridium difficile CD196. Infect Immun 65, 1402–1407.
Stubbs S, Rupnik M, Gibert M, Brazier J, Duerden B and Popoff M. (2000) Production of actin-specific ADP-ribosyltransferase (binary toxin) by strains of Clostridium difficile. FEMS Microbiol Lett 186, 307–312.
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This work was supported by grant J3-0194-0377-08.
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Rupnik, M. (2010). Clostridium difficile Toxinotyping. In: Mullany, P., Roberts, A.P. (eds) Clostridium difficile. Methods in Molecular Biology™, vol 646. Humana Press. https://doi.org/10.1007/978-1-60327-365-7_5
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DOI: https://doi.org/10.1007/978-1-60327-365-7_5
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