The study aimed to characterize the genome of a Colombian strain of Leptospira santarosai to infer bacterial virulence factors. Under the approach of a quantitative research, an isolate of Colombian origin of L. santarosai was sequenced using new generation 454 GS FLX Titanium sequencers. Subsequently, bioinformatics programs were used for the physical description of the genome (number of genes, size), open reading frame prediction, prediction of proteins and their orthologs in other pathogenic species, of intermediate pathogenicity and non-pathogenic, cellular localization of proteins. The assembly of the spirochete genome was achieved Leptospira santarosai serogroup Autumnalis serovar Aliceand the description of the genetic, structural and functional characteristics of its genes. It is concluded that the core- genome of the isolate is composed of 1747 proteins, which are common to all L. santarosai strains available in GenBank. It was determined that it has a total of 4,138 proteins, 141 of which are unique to its genome. The possible role of the virulence factors found in the Colombian isolate was identified and described. The study contributes to the understanding of the pathophysiological mechanisms induced by Leptospira .