Rapid and direct detection of Clostridium chauvoei by PCR of the 16S-23S rDNA spacer region and partial 23S rDNA sequences

Clostridium chauvoei causes blackleg, which is difficult to distinguish fro m the causative clostridia of malignant edema. Therefore, a single-step PCR system was developed for specific detection of C. chauvoei DNA using prime rs derived from the 168-23S rDNA spacer region and partial 23S rDNA sequenc es. The specificity of the single-step PCR system was demonstrated by testi ng 37 strains of clostridia and 3 strains of other genera. A 509 bp PCR pro duct, which is a C. chauvoei-specific PCR product, could be amplified from all of the C. chauvoei strains tested, but not from the other strains. More over, this single-step PCR system specifically detected C. chauvoei DNA in samples of muscle from mice 24 hr after inoculation with 100 spores of C. c hauvoei, and in clinical materials from a cow affected with blackleg. These results suggest that our single-step PCR system may be useful for direct d etection of C. chauvoei in culture and in clinical materials from animals a ffected with blackleg.