title: Antigen recognition by serum antibodies in white-tailed deer (Odocoileus virginianus) experimentally infected with Mycobacterium bovis.
abstract: White-tailed deer (Odocoileus virginianus) have emerged as reservoirs of bovine tuberculosis in northern America. For tuberculosis 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 noninfected deer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for immunoglobulin specific to M. bovis antigens. Various routes of experimental M. bovis infection, such as intratonsillar inoculation (n = 11), aerosol (n = 6), and exposure to infected deer (in contact, n = 8), were studied. Upon infection, specific bands of reactivity at approximately 24 to 26 kDa, approximately 33 kDa, approximately 42 kDa, and approximately 75 kDa to M. bovis whole-cell sonicate were detected by immunoblot. Lipoarabinomannan-specific immunoglobulin was detected as early as 36 days postchallenge, and responses were detected for 94% of intratonsillarly and "in-contact"-infected deer. In MAPIA, sera were tested with 12 native and recombinant antigens coated on nitrocellulose. All in-contact-infected (8 of 8) and 10 of 11 intratonsillarly 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, three of six deer receiving a very low dose of M. bovis via aerosol exposure produced antibody specific to one or more recombinant proteins. M. bovis was isolated from one of three nonresponding 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.
authors.affiliation: National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, Bacterial Diseases of Livestock Research Unit, Ames, IA 50010-0070, USA. rwaters@nadc.ars.usda.gov
authors.forename: W R
authors.initials: WR
authors.lastname: Waters
authors.forename: M V
authors.initials: MV
authors.lastname: Palmer
authors.forename: J P
authors.initials: JP
authors.lastname: Bannantine
authors.forename: D L
authors.initials: DL
authors.lastname: Whipple
authors.forename: R
authors.initials: R
authors.lastname: Greenwald
authors.forename: J
authors.initials: J
authors.lastname: Esfandiari
authors.forename: P
authors.initials: P
authors.lastname: Andersen
authors.forename: J
authors.initials: J
authors.lastname: McNair
authors.forename: J M
authors.initials: JM
authors.lastname: Pollock
authors.forename: K P
authors.initials: KP
authors.lastname: Lyashchenko
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000917:Antibody Formation; D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial / Q000276:immunology; D003670:Deer; D007118:Immunoassay / Q000662:veterinary; D008070:Lipopolysaccharides / Q000276:immunology; D009164:Mycobacterium Infections / Q000175:diagnosis* / Q000662:veterinary; D009163:Mycobacterium bovis / Q000276:immunology*; D012698:Serologic Tests / Q000662:veterinary
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D008070:Lipopolysaccharides; C050016:lipoarabinomannan