Desarrollo y validación de dos inmunoensayos para la detección de Brucella canis en perros

Canine Brucellosis is cause of reproductive failure and early detection is still a challenge in infected dogs. The aim of present study was to test two antigens to be used in two different indirect immunoassays for the detection of Brucella canis infection in dogs and in-house validation was performed: one from a crude sonicated antigen of Brucella canis RM6/66 (CA–iELISA) and another using the immunodominant protein GroEL (GroEL–iELISA). The cutoff value was determined, using sera from infected dogs; then, a reproducibility test was done in which CA-iELISA had a coefficient of variation (CV) below 15 % for negatives and positives, while for GroEL-iELISA was above 15 %. Robustness was then tested, with significant differences in the GroEL-iELISA results, but not in CA-iELISA. Selectivity test revealed a cross-reaction to Leptospira interrogans with CA-iELISA, and L. interrogans and Salmonella spp. with GroEL-iELISA. Sample stability tests showed that the samples should be kept at room temperature for no more than 2 h, at 4 °C for no more than 24 h and could be kept at -20 °C for up to 30 days. Finally, the sensitivity and specificity of each test was calculated, resulting in 100 % for both CA-iELISA and a sensitivity of 44 % and a specificity of 67 % for GroEL-iELISA. In conclusion, CA-iELISA has been found to be better than GroEL-iELISA detecting Brucella canis.