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Mycobacterium bovis  infection has been described in many wildlife species across Africa. However, diagnostic tests are lacking for many of these, including warthogs ( Phacochoerus africanus ). Most literature on suids has focused on using serological tools, with few studies investigating the use of cell-mediated immune response (CMI) assays. A recent study showed that warthogs develop measurable CMI responses, which suggests that cytokine gene expression assays (GEAs) may be valuable for detecting  M .  bovis -infection, as shown in numerous African wildlife species. Therefore, the aim of the study was to develop GEAs capable of distinguishing between  M .  bovis -infected and uninfected warthogs. Whole blood was stimulated using the QuantiFERON-TB Gold (In-Tube) system, using ESAT-6 and CFP-10 peptides, before determining the relative gene expression of five reference ( B2M ,  H3F3A ,  LDHA ,  PPIA  and  YWHAZ ) and five target ( CXCL9 ,  CXCL10 ,  CXCL11 ,  IFNG  and  TNFA ) genes through qPCR. The reference gene  H3F3A  was the most stably expressed, while all target genes were significantly upregulated in  M .  bovis -infected warthogs with the greatest upregulation observed for  CXCL10 . Consequently, the  CXCL10  GEA shows promise as an ante-mortem diagnostic tool for the detection of  M .  bovis -infected warthogs.

Cytokine gene expression assay as a diagnostic tool for detection of Mycobacterium bovis infection in warthogs (Phacochoerus africanus)