Use of Polymerase Chain Reaction and Endonuclease Restriction BccI for the Differential Leptospira interrogans Detection

Leptospirosis is a world disease, affecting animals and humans, which is transmitted by all infected animal, having the rodents as the most recognized. Leptospira interrogans serovar canicola, Leptospira interrogans serovar icterohaemorragiae, Leptospira interrogans serovar pomona and Leptospira interrogans serovar grippotyphosa affect canines, pigs, horses and cattle. A differential diagnosis method of four Leptospira serovars is our objective, through the molecular technique of the Polymerase Chain Reaction (PCR) adding the action of the restriction enzyme BccI, and thus generate fragments of different size for the recognition of the gene encoding a lipoprotein membrane LipL32. As conclusion, the detection of Leptospira interrogans by PCR was possible by that the primers designed for the realization of the method are specific for this agent. The differentiation of the 4 serovars of Leptospira was successful because the restriction enzyme cleaves the double strand of DNA in the 4 serovars, delivering expected results.