Toxintyping of Clostridium perfringens Strains by Colony Multiplex PCR

Abstract:

Four primers against the genes encoding (cpa, cpb, etx, and iA) four major toxins(α, β, ε, ι) of Cl. perfringens were designed and the colony multiplex PCR of identification and genotyping of Cl. perfringens strains were developed. Cl. perfringens reference strains stored in china institute of veterinary drug control including A, B, C, D and E genotyping were genotyped using the colony muitiplex PCR assay. The expected sequences were obtained successfully by the colony multiplex PCR assay. But the sequences were not obtained from Cl. novyi, Cl. septicum and Cl. tetani. The expected sequences were obtained from Cl. perfringens individual colony diluted to 100 times with 0.85% saline solution.13 Cl. perfringens strains isolated from diferent animals were genotyped using the colony multiplex PCR assay, and the results were comparaed with the results of toxins neutranization test in mice. The two assays showed good accordance. These results showed that the development of the colony multiplex PCR is very important for early and fast identification and genotyping of Cl. perfringens in china.