DETECTION OF ALFA, BETA AND EPSILON TOXINS GENES IN CLOSTRIDIUM PERFRINGENS STRAINS ISOLATED FROM BOVINES BY THE POLIMERASE CHAIN REACTION

ABSTRACT

The Clostridium perfringens, an aerobic microorganism, is present in soil and intestinal tracts of mammals. It causes gaseous gangrene and food intoxication in human beings and enterotoxemic diseases in domestic animals.C. perfringens is classified into 5 types (A,B,C,D and E) according to the production of four main toxins: (alfa-α, beta-β, gama-γ, delta-δ e épsilon-ε). The identification of these toxins is made through serum neutralization test in animals, using specific anti-sera, however these reagents are of high cost and difficult to be obtained from international reference laboratories. To aid in the typing of the C. perfringens strains in our environment, the present work aimed to assess the use of the polymerase chain reaction (PCR) for the detection of the genes coding the alpha (cpa), beta (cpb) and epsilon (etx) toxins in bovines isolates.The analytical sensitivity of the PCR technique, standardized from the total bacterial DNA, was 2.27 ng/?L for the cpa gene and 227 pg/?L for the cpb and etx genes. Among the 35 C. perfringens isolates typed by PCR, 16 (45.7%) were of type A, 18 (51.4%) of type C, and 1 (2.9%) belonged to type B.

KEY WORDS
Clostridium perfringens ; typing; bovines; PCR