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Title:   Author  item
                                            Waters, Wadeitem
                                            Palmer, Mitchellitem
                                            Bannantine, Johnitem
                                            Whipple, Dianaitem
                                            GREENWALD, R - CHEMBIO DIAG. SYS. INCitem
                                            ESFANDIARI, J - CHEMBIO DIAG. SYS. INC.item
                                            ANDERSEN, P - STATENS SERUM INSTITUTitem
                                            MCNAIR, J - BELFAST, UKitem
                                            POLLOCK, J - BELFAST, UKitem
                                            LYASHCHENKO, K - STATENS SERUM INSTITUT
  Submitted to: Clinical and Diagnostic Laboratory Immunology
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 7/13/2004
                                    Publication  Date: 9/1/2004
                                    Citation: Waters, W.R., Palmer, M.V., Bannantine, J.P., Whipple, D.L., Greenwald, R., Esfandiari, J., Andersen, P., Mcnair, J., Pollock, J.M., Lyashchenko, K.P. 2004. Antigen recognition by serum antibodies in white-tailed deer (odocoileus virginianus) experimentally infected with mycobacterium bovis. Clinical and Diagnostic Laboratory Immunology. 11(5):849-55.Interpretive Summary: White-tailed deer are wildlife reservoirs of bovine tuberculosis within the United States. The presence of this reservoir host seriously threatens ongoing efforts to eradicate this disease from cattle. Captive deer, in addition to wild deer, may also become infected with this agent. Thus, it is of interest to develop tests to detect infected white-tailed deer. In the present study, it was determined that deer experimentally infected with tuberculosis produce antibodies to the bacterium and that these antibodies are detectable by simple laboratory methods. Since this technology is easily transferred to diagnostic laboratories, serological assays may prove practical for use in detecting tuberculosis infection in captive white-tailed deer. These findings will be useful for the development of improved methods of bovine tuberculosis control, thus, benefiting the cattle industry.
                                Technical Abstract: White-tailed deer (Odocoileus virginianus) have emerged as reservoirs of bovine tuberculosis (TB) in Northern America. For TB surveillance of captive 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), by 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 postchallenge 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. M. bovis was not isolated from 2/3 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.


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