Identification of the virB operon genes encoding the type IV secretion system, in Colombian Brucella canis isolates

Canine brucellosis is a zoonotic disease caused by Brucella canis. The establishment of intracellular replicative niches of B. canis is mediated by proteins secreted by the type IV secretion system, which is encoded by the virB operon. The characterization of such genes has been conducted in other species of the genus, but not in B. canis.We report the design of a multiplex PCR test for the detection of the virB operon genes of B. canis. Primers for each of the 12 genes were designed and evaluated using bioinformatics tools. A multiplex PCR assay was standardized and applied to 36 isolates obtained from infected dogs of Aburrá Valley kennels, as well to the Brucella abortus, Brucella melitensis, Brucella suis and Brucella ovis DNA strains. As a result of the in silico design, a pair of primers for each gene was selected. All species and isolates evaluated showed evidence of the presence of the entire virB operon.