WFL Publisher

A simple mPCR procedure was used to identify four toxitypes of Clostridium perfringens collected from different origins. Eighteen strains of Clostridium perfringens were identified and typed by classical methods (dermonecrotic method in guinea pigs and seroneutralization test in mice). All the strains were analyzed by PCR using gene of toxin alpha, gene of toxin beta, gene of toxin epsilon and gene of toxin iota. The results reveal a toxin gene in 13 (72.22%) strains of Clostridium perfringens, only 11 (61.11%) strains were identified previously as type A by classical method, as well as 3 strains (16.67%) were identified as type C and 1 strain (5.56%) was identified as D by PCR typing. Moreover, mPCR results confirmed the traditional methods in typing one strain as type B (5.56%). Also, mPCR method can detect 2 other strains of type A directly in the feces and intestinal contents of the examined chicken which gave negative results in bacteriological examination. Thus, PCR technique can become a first-choice tool for the identification and typing of the Clostridium perfringens strains which initiate enteric disease. In turn, this would simplify the development of vaccines adapted to the epidemiological situation. Taken all together, PCR method is easy, time saving and applicable to differentiate Clostridium perfringens types as an alternative to animal tests