Molecular epidemiology survey of toxinogenic Clostridium perfringens strain types by multiplex PCR

Toxin genotypes of 95 C. perfringens strains collected within a 45-year per iod were analysed by a multiplex PCR, A set of primers designed for 4 diffe rent genes encoding the alpha, beta, epsilon, and iota toxins was used in a single reaction with a sensitivity of gene detection of 200 fg for DNA ext racted from pure culture. Most of the strains (97%) conformed to the A biot ype, and the remaining to the C or E biotypes. For biotype determination, s eroneutralization of lethality in mice was performed by intravenous injecti on. Toxin phenotype and penotype profile were concordant in 94% of strains. Our results documented the presence of rare toxin genotypes of C. perfring ens in a Polish geographical region and indicated the suitability of multip lex PCR as a method supplementing classical techniques and providing better insight into the prevalence of toxinogenic C. perfringens strains.