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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.    Conclusion  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.

Evaluation of ethanol vortex ELISA for detection of bovine tuberculosis in cattle and deer