title: Use of ethanol extract of Mycobacterium bovis for detection of specific antibodies in sera of farmed red deer (Cervus elaphus) with bovine tuberculosis.
abstract: BACKGROUND
  Bovine tuberculosis (bTB) in wildlife species poses a threat to domestic livestock in many situations. Control programs for bTB in livestock depend on testing and slaughtering the positive animals; however, the currently available diagnostic tests often have poor specificity. In our previous study, we developed a specific and sensitive enzyme linked immunosorbent assay (ELISA) for another mycobacterial disease - Johne's disease, using surface antigens of Mycobacterium avium ssp. paratuberculosis (MAP) extracted by briefly agitating the bacilli in 80% ethanol solution. The ELISA test was named ethanol vortex ELISA (EVELISA). The objective of this study is to examine whether EVELISA technique could be used to specifically detect anti-Mycobacterium bovis (M. bovis) antibodies in the serum of M. bovis-infected farmed red deer (Cervus elaphus). We tested a total of 45 red deer serum samples, divided in 3 groups - uninfected animals (n = 15), experimentally infected with M. bovis (n = 15) and experimentally infected with MAP (n = 15).
  
  
  RESULTS
  The presence of anti-M. bovis antibodies was tested using an ethanol extract of M. bovis. Without absorption of anti-MAP cross reactive antibodies, it was found that 13 out of the 15 MAP-infected animals showed high antibody binding. Using heat killed MAP as an absorbent of cross reactive antibodies, anti-M. bovis antibodies were detected in 86.7% of M. bovis-infected animals with minor false positive results caused by MAP infection.
  
  
  CONCLUSIONS
  The results from this study suggest that EVELISA may form a basis for a sensitive and specific test for the diagnosis of bTB in farmed red deer.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Wadhwa
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Johnson
authors.forename: Colin G
authors.initials: CG
authors.lastname: Mackintosh
authors.forename: J Frank T
authors.initials: JF
authors.lastname: Griffin
authors.forename: W Ray
authors.initials: WR
authors.lastname: Waters
authors.forename: John P
authors.initials: JP
authors.lastname: Bannantine
authors.affiliation: Department of Forestry, Wildlife and Fisheries, Center for Wildlife Health, University of Tennessee Institute of Agriculture, Knoxville, Tennessee 37996, USA. seda@utk.edu.
authors.forename: Shigetoshi
authors.initials: S
authors.lastname: Eda
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D002417:Cattle; D003670:Deer / Q000382:microbiology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D016927:Mycobacterium avium subsp. paratuberculosis / Q000276:immunology; D009163:Mycobacterium bovis / Q000276:immunology*; D014380:Tuberculosis, Bovine / Q000175:diagnosis* / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial