title: Evaluation of ethanol vortex ELISA for detection of bovine tuberculosis in cattle and deer.
abstract: BACKGROUND
  The use of serological assays for diagnosis of bovine tuberculosis (TB) has been intensively studied and use of specific antigens have aided in improving the diagnostic accuracy of the assays. In the present study, we report an in-house enzyme linked immunosorbent assay (ELISA), developed by using ethanol extract of Mycobacterium bovis (M. bovis). The assay, named (ethanol vortex ELISA [EVELISA]), was evaluated for detection of anti- M. bovis antibodies in the sera of cattle and white-tailed deer.
  
  
  METHODS
  By using the EVELISA, we tested sera obtained from two species of animals; cattle (n = 62 [uninfected, n = 40; naturally infected, n = 22]) and white-tailed deer (n = 41 [uninfected, n = 25; naturally infected, n = 7; experimentally infected, n = 9]). To detect species specific molecules, components in the ethanol extract were analyzed by thin layer chromatography and western blotting.
  
  
  RESULTS
  Among the tested animals, 77.2% of infected cattle and 87.5% of infected deer tested positive for anti- M. bovis antibody. There were only minor false positive reactions (7.5% in cattle and 0% in deer) in uninfected animals. M. bovis -specific lipids and protein (MPB83) in the ethanol extract were detected by thin layer chromatography and western blotting, respectively.
  
  
  CONCLUSIONS
  The results warrant further evaluation and validation of EVELISA for bovine TB diagnosis of traditional and alternative livestock as well as for free-ranging animal species.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Wadhwa
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Johonson
authors.forename: Keiko
authors.initials: K
authors.lastname: Eda
authors.forename: W Ray
authors.initials: WR
authors.lastname: Waters
authors.forename: Mitchell V
authors.initials: MV
authors.lastname: Palmer
authors.forename: John P
authors.initials: JP
authors.lastname: Bannantine
authors.affiliation: Center for Wildlife Health, Department of Forestry, Wildlife and Fisheries, University of Tennessee Institute of Agriculture, 274 Ellington Plant Science Building, Knoxville, TN 37996, USA. seda@utk.edu.
authors.forename: Shigetoshi
authors.initials: S
authors.lastname: Eda
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D002417:Cattle; D003670:Deer / Q000382:microbiology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D000431:Ethanol / Q000737:chemistry*; D005189:False Positive Reactions; D009163:Mycobacterium bovis / Q000302:isolation & purification*; D012698:Serologic Tests / Q000379:methods / Q000662:veterinary; D014376:Tuberculosis / Q000097:blood / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*
chemical_list: D000907:Antibodies, Bacterial; D000431:Ethanol