IDENTIFICATION OF CLOSTRIDIUM-PERFRINGENS BY 16S AND 23S RIBOSOMAL-RNA OLIGONUCLEOTIDE PROBES

The published 16S rRNA sequence of Clostridium perfringens [Garnier et al. (1991] was compared to the 16S rRNA database of Clostridium speci es and two C. perfringens-specific oligonucleotide probes were develop ed. In addition, the 23S rDNA of C. perfringens was partially sequence d and used to design a third oligonucleotide probe. The specificity of the probes was determined by dot blot hybridisation using crude rRNA from several strains of each C. perfringens type, Gram-positive repres entatives of various genera and Escherichia coli. All probes were spec ific for C. perfringens but one 16S rRNA probe failed to detect all of the C. perfringens strains tested. Using the polymerase chain reactio n technology in combination with a nonradioactive hybridisation assay the identification of isolated DNA from pure cultures of C. perfringen s that express the whole range of C. perfringens toxin types was achie ved within a few hours.