Abstract
Brucellosis in livestock significantly impacts many countries’ economies, primarily affecting animal health and productivity. It causes direct losses for the animal industry and is a barrier to the international trade of live animals. It is a zoonosis as well, hence affecting public health. A newly developed multiplex high-resolution melt (HRM)real-time (q) polymerase chain reaction (PCR) for simultaneous detection of four abortive zoonotic agents in domestic ruminants was compared with probe-based qPCR assaysto detect Brucella spp. and Coxiella burnetii. Ninety-seven (97) clinical samples from cattle, sheep, and goat abortion cases submitted to Botswana National Veterinary Laboratory from 2010 to 2021 for disease diagnosis, and 34 milk samples submitted to the same laboratory for routine quality control were subjected to fourplex HRM and Taqman assays. The fourplex assay detected Brucella spp. in 13 samples and co-infection of Brucella spp. with Coxiella burnetii in 45 samples.There was a perfect agreement between the HRM assay and each of the two qPCRs (kappa =1). Similarly, the Bland-Altman analysis showed a mean Cq difference of -0.7 for HRM versus Brucella qPCR and 0.63 for HRM versus Coxiella qPCR, suggesting a good agreement. In conclusion, the HRM fourplex assay is a valuable diagnostic tool to detect Brucella species with reliable Cq values com-parable with the Taqman assays.References
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