Serum PCR Diagnosis of Brucella melitensis Infection in Rev. 1 Vaccinated Sheep

Serological diagnosis provides a robust and effective approach to monitoring and controlling small ruminant brucellosis. Brucella melitensis Rev. 1 is a live vaccine strain used in prophylactic vaccination against small ruminant brucellosis. Because the vaccine strain shares identical serological antigens with the corresponding field strains, differentiating infected from vaccinated animals (DIVA) serological responses hamper surveillance campaigns and interventions that involve vaccination. We have developed a serum PCR-based approach in which we amplify and sequence Brucella omp2a as a DIVA solution and tRNA (uracil-5-)-methyltransferase as a species marker in the serum samples to determine the etiological agent involved in brucellosis field cases. Using this method, we identified the involvement of both the Rev. 1 vaccine strain and a field strain in an outbreak of brucellosis in a flock. This method represents a novel approach in studying the etiology of brucellosis using serum samples as a source of the pathogen’s DNA.