Abstract

A simple polymerase chain reaction (PCR) procedure was used to identify four toxitypes of Clostridium perfringens collected from different origins. Eighteen (18) strains of C. perfringens were identified and typed by classical methods (dermonecrotic method in guinea pigs and sero-neutralization 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 α toxin gene in 13 (72.22%) strains of C. perfringens; only 11 (61.11%) strains of them were identified previously as type A by classical method, three strains (16.67%) were identified as type C and one strain (5.56%) was identified as type D by PCR. Moreover, PCR results confirmed the traditional methods in typing one strain as type B (5.56%). Also, PCR method could detect two 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 C. perfringens 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 C. perfringens types as an alternative to animal tests.

Key words: Clostridium perfringens, major toxins, polymerase chain reaction (PCR)typing, enteric disease.