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

BMC Veterinary Research

Shigetoshi Eda¹  John P Bannantine²  Mitchell V Palmer²  W Ray Waters²  Keiko Eda¹  Rachel E Johonson¹  Ashutosh Wadhwa¹ 

[1] 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;Infectious Bacterial Diseases of Livestock Research Unit, National Animal Disease Center, USDA Agricultural Research Service, Ames, IA 50010, USA

关键词: EVELISA;    Mycobacterium bovis;    ELISA;    White-tailed deer;    Cattle;    Bovine tuberculosis;   

Others  :  865480
DOI  :  10.1186/1746-6148-10-147

 received in 2013-11-22, accepted in 2014-06-20,  发布年份 2014

【 摘 要 】

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.

【 授权许可】

   
2014 Wadhwa et al.; licensee BioMed Central Ltd.

【 图 表 】

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