Use of the MILLIPLEX® bovine cytokine/chemokine multiplex assay to identify Mycobacterium bovis-infection biomarkers in African buffaloes (Syncerus caffer)

As a recognized Mycobacterium bovis maintenance host, the African buffalo (Syncerus caffer) poses transmission risks to livestock, humans and other wildlife. Early detection of M. bovis infection is critical for limiting its spread. Currently, tests detecting cell-mediated immune responses are used for diagnosis in buffaloes. However, these may have suboptimal sensitivity or specificity, depending on the blood stimulation method. Recent evidence suggests that assays using combinations of host cytokine biomarkers may increase diagnostic performance. Therefore, this study aimed to investigate the application of a MILLIPLEX (R) bovine cytokine/chemokine multiplex assay to identify candidate biomarkers of M. bovis infection in buffaloes. Whole blood from twelve culture-confirmed M. bovis-infected buffaloes, stimulated with the QuantiFERON((R)) TB Gold Plus in-tube system, was tested using the MILLIPLEX (R) platform. Results indicated binding of bovine antibodies to fifteen buffalo cytokine/chemokine targets. Moreover, there was a significant difference in concentrations between unstimulated and TB antigen-stimulated buffalo samples for seven cytokines/chemokines included in the kit. Although these preliminary results require further investigation in larger sample sets and a comparison between M. bovis-infected and uninfected cohorts, the utility of the MILLIPLEX (R) platform in a novel species was demonstrated, in addition to identifying potential African buffalo cytokines for future research.

Automated summary

The African buffalo as a maintenance host for Mycobacterium bovis poses transmission risks to livestock, humans, and other wildlife. Using the MILLIPLEX(R) platform to test whole blood samples from infected buffaloes revealed potential cytokine biomarkers and demonstrated the utility of the platform in this novel species. Further investigation is needed to confirm these preliminary findings.