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Journal of Clinical Microbiology, May 2004, p. 1933-1939, Vol. 42, No. 5
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.5.1933-1939.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Prevalence and Characterization of a Binary Toxin (Actin-Specific ADP-Ribosyltransferase) from Clostridium difficile

Carina Gonçalves,1 Dominique Decré,1,2* Frédéric Barbut,1,2,3 Béatrice Burghoffer,1 and Jean-Claude Petit1,2

UPRES EA2392, Faculté de Médecine, UFR Saint-Antoine, Université Paris 6,1 Laboratoire de Bactériologie,2 Unité d'Hygiène et de Lutte contre les Infections Nosocomiales (UHLIN), Hôpital Saint-Antoine, Assistance Publique-Hôpitaux de Paris, Paris, France3

Received 13 June 2003/ Returned for modification 30 July 2003/ Accepted 21 January 2004

In addition to the two large clostridial cytotoxins (TcdA and TcdB), some strains of Clostridium difficile also produce an actin-specific ADP-ribosyltransferase, called binary toxin CDT. We used a PCR method and Southern blotting for the detection of genes encoding the enzymatic (CDTa) and binding (CDTb) components of the binary toxin in 369 strains isolated from patients with suspected C. difficile-associated diarrhea or colitis. Twenty-two strains (a prevalence of 6%) harbored both genes. When binary toxin production was assessed by Western blotting, 19 of the 22 strains reacted with antisera against the iota toxin of C. perfringens (anti-Ia and anti-Ib). Additionally, binary toxin activity, detected by the ADP-ribosyltransferase assay, was present in only 17 of the 22 strains. Subsequently, all 22 binary toxin-positive strains were tested for the production of toxins TcdA and TcdB, toxinotyped, and characterized by serogrouping, PCR ribotyping, arbitrarily primed PCR, and pulsed-field gel electrophoresis. All binary toxin-positive strains also produced TcdB and/or TcdA. However, they had significant changes in the tcdA and tcdB genes and belonged to variant toxinotypes III, IV, V, VII, IX, and XIII. We could differentiate 16 profiles by using typing methods, indicating that most of the binary toxin-positive strains were unrelated.

* Corresponding author. Mailing address: Laboratoire de Bactériologie, CHU Saint-Antoine, 27 rue Chaligny, 75012 Paris cedex 12, France. Phone: 33-1-40-01-13-82. Fax: 33-1-40-01-14-27. E-mail: dominique.decre{at}chusa.jussieu.fr.

Journal of Clinical Microbiology, May 2004, p. 1933-1939, Vol. 42, No. 5
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.5.1933-1939.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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