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

Document Type

Article

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.