Development and validation of two immunoassays for the detection of Brucella canis in dogs
Keywords:
Brucella canis, GroEL, ELISAAbstract
Canine brucellosis is a cause of reproductive failure and early detection in infected dogs remains a challenge. The aim of the present study was to test two antigens to be used in two different immunoassays for the detection of Brucella canis infection in dogs: one from Brucella canis RM6/66 sonicated crude antigen (CA-iELISA) and the other using the immunodominant protein GroEL (GroEL-iELISA). The cut-off point was determined using sera from infected dogs; subsequently, reproducibility was measured and a coefficient of variation (CV) below 15 % was obtained for negatives and positives with the CA-iELISA, whereas, for the GroEL-iELISA they were higher than 15 %. In the robustness test, there were significant differences for the GroEL-iELISA, but not for the CA-iELISA. The selectivity test showed cross-reactivity with Leptospira interrogans in the CA-iELISA, and with L. interrogans and Salmonella spp. in the GroEL-iELISA. From the sample stability tests, it was demonstrated that samples should be stored at room temperature for no more than 2 hours, at 4 ºC for no more than 24 hours and can be kept at -20 ºC for up to 30 days. Finally, sensitivity and specificity were calculated, resulting in 100 % for both CA-iELISA; sensitivity of 44 % and specificity of 67 % for GroEL-iELISA. In conclusion, the CA-iELSA proved to be better at detecting Brucella canis than the GroEL-iELISA.
http://dx.doi.org/10.21929/abavet2021.31
e2020-90.
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