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Clostridium perfringens  enterotoxin (CPE) is a major virulence factor for human gastrointestinal diseases, such as food poisoning and antibiotic associated diarrhea. The CPE-encoding gene ( cpe ) can be chromosomal or plasmid-borne. Recent development of conventional PCR  cpe -genotyping assays makes it possible to identify  cpe  location (chromosomal or plasmid) in type A isolates. Initial studies for developing  cpe  genotyping assays indicated that all  cpe -positive strains isolated from sickened patients were typable by  cpe -genotypes, but surveys of  C. perfringens  environmental strains or strains from feces of healthy people suggested that this assay might not be useful for some  cpe -carrying type A isolates. In the current study, a pulsed-field gel electrophoresis Southern blot assay showed that four  cpe -genotype untypable isolates carried their  cpe  gene on a plasmid of ∼65 kb. Complete sequence analysis of the ∼65 kb variant  cpe -carrying plasmid revealed no intact IS elements and a disrupted cytosine methyltransferase ( dcm ) gene. More importantly, this plasmid contains a conjugative transfer region, a variant  cpe  gene and variant iota toxin genes. The toxin genes encoded by this plasmid are expressed based upon the results of RT-PCR assays. The ∼65 kb plasmid is closely related to the pCPF4969  cpe  plasmid of type A isolates. MLST analyses indicated these isolates belong to a unique cluster of  C. perfringens . Overall, these isolates carrying a variant functional  cpe  gene and iota toxin genes represent unique type E strains.

Identification of Novel Clostridium perfringens Type E Strains That Carry an Iota Toxin Plasmid with a Functional Enterotoxin Gene