Use of ethanol extract of Mycobacterium bovis for detection of specific antibodies in sera of farmed red deer (Cervus elaphus) with bovine tuberculosis

Authors

Ashutosh Wadhwa, University of Tennessee Institute of Agriculture, Knoxville
Rachel E. Johnson, University of Tennessee Institute of Agriculture, Knoxville
C. G. Mackintosh, Invermay Agricultural Research Centre, Mosgiel, New ZealandFollow
J Frank Griffin, University of Otago, Dunedin
W Ray Waters, USDA-ARS
John Bannantine, USDA-ARSFollow
Shigetoshi Eda, University of Tennessee Institute of Agriculture, KnoxvilleFollow

Date of this Version

2013

Citation

Wadhwa et al. BMC Veterinary Research 2013, 9:256 http://www.biomedcentral.com/1746-6148/9/256

Comments

2013 Wadhwa et al.

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.