Abstract

White-tailed deer (Odocoileus virginianus) have emerged as reservoirs of bovine tuberculosis (TB) in Northern America. For TB surveillance of deer, antibody-based assays are particularly attractive because deer are handled only once, and immediate processing of the sample is not required. Sera collected sequentially from 25 Mycobacterium bovis-infected and 7 non-infected deer were evaluated by ELISA, immunoblotting, and multi-antigen print immunoassay (MAPIA) for immunoglobulin specific to M. bovis antigens. Various routes of experimental M. bovis infection, such as intratonsilar inoculation (n=11), aerosol (n=6), and exposure to infected deer (in contact, n=8) were studied. Upon infection, specific bands of reactivity at ∼24–26 kDa, ∼33 kDa, ∼42 kDa, and ∼75 kDa to M. bovis whole cell sonicate were detected by immunoblot. Lipoarabinomannan-specific immunoglobulin was detected as early as 36 days post challenge, and responses were detected for 94% of intratonsilar and in contact infected deer. In MAPIA, sera were tested with 12 native and recombinant antigens coated on nitrocellulose. All “in contact” infected (8/8) and 10/11 intratonsilarly-infected deer produced antibody reactive with one or more of the recombinant/native antigens. Responses were boosted by injection of tuberculin for intradermal tuberculin skin testing. Additionally, 3/6 deer receiving a very low dose of M. bovis via aerosol exposure produced antibody specific to one or more recombinant proteins. Mycobacterium bovis was isolated from 1/3 non-responding aerosol-challenged deer. Of the 12 antigens tested, the most immunodominant protein was MPB83; however, a highly sensitive serodiagnostic test will likely require use of multiple antigens.