Optimized interferon-gamma release assays for detection of Mycobacterium bovis infection in African buffaloes (Syncerus caffer)

Abstract The African buffalo (Syncerus caffer) is an economically and ecologically important wildlife species in South Africa; it is also a primary wildlife maintenance host of Mycobacterium bovis. Accurate and early detection of M. bovis infection in buffaloes is important for controlling transmission. Assays that detect cell-mediated immune responses to M. bovis in buffaloes have been developed although these often display suboptimal sensitivity or specificity. Therefore, the aim of this study was to evaluate the newly available Mabtech bovine interferon-gamma (IFN-γ) ELISAPRO kit and optimize its use for detection of buffalo IFN-γ in whole blood samples stimulated with the QuantiFERON® TB Gold Plus antigens. Additionally, the test performance of the Mabtech IFN-γ release assay (IGRA) was compared to the currently used Cattletype® IGRA by determining buffalo-specific cut-off values for the two IGRAs and using gold standard-positive (M. bovis culture-confirmed) and M. bovis-unexposed negative cohorts. Validation of the Mabtech ELISA revealed negligible matrix interference and a linear and parallel response for recombinant bovine and native buffalo IFN-γ in the range 1.95 to 250 pg/mL. Intra- and inter-assay reproducibility produced coefficients of variation