Journal of Veterinary and Animal Sciences

Volume: 55 Issue: 2

  • Open Access
  • Short Communication

Evaluation of a Dot ELISA for tuberculosis screening in Asian elephants (Elephas maximus)

David Abraham1*, Chintu Ravishankar2, P.M. Deepa1, K. Sumod2, K.C. Bipin1, C.V. Rajani3 and K. Vijayakumar1

1. Department of Veterinary Epidemiology and Preventive Medicine

2. Department of Veterinary Microbiology

3. Department of Veterinary Anatomy

College of Veterinary and Animal Sciences, Pookode, Wayanad- 673 576 Kerala Veterinary and Animal Sciences University Kerala, India.

*Corresponding author: [email protected], Ph. 9446536380

Year: 2024, Page: 465-470, Doi: https://doi.org/10.51966/jvas.2024.55.2.465-470

Received: Dec. 19, 2023 Accepted: March 30, 2024 Published: June 30, 2024

Abstract

Banked serum samples of 60 dead captive Asian elephants were used to evaluate a dot ELISA for tuberculosis screening, of which 30 each were taken as cases and controls. The 30 samples that were recorded as positive for Mycobacterium tuberculosis by culture and PCR besides being positive on the DPP® Vet-TB Elephant Assay (DPP) and based on lesions during post-mortem examinations were utlised as known positive controls for this study. The 30 controls were tested negative on DPP and were from elephants that had not shown post-mortem lesions. The dot ELISA was designed on a nitrocellulose membrane template, using Protein A- Horseradish peroxidase as the conjugate and diaminobenzidine tetrahydrochloride as the chromogen substrate. Standardisation of the assay was done using a purified M. tuberculosis antigen, ESAT6-CFP10. Pooled sera samples from live elephants tested recently on the DPP were taken as positive and negative controls. Test results of the dot ELISA were evaluated by comparing the results with the disease status of the cases and controls based on the ante-mortem DPP results and post-mortem lesions. Diagnostic sensitivity (66.67%), specificity (93.33%), positive (90.91%) and negative (73.68%) predictive values were then estimated for the dot ELISA.

Keywords: Tuberculosis, elephant, dot ELISA, ESAT6-CFP10

References

Abraham, D., Cheeran, J. V., Sukumar, R., Mikota, S. K., Rao, S., Ganguly, S. and Varma, S. 2008. Health assessment of captive Asian elephants in India with special reference to tuberculosis, A preliminary report submitted to Project Elephant, Ministry of Environment and Forests, Government of India. Asian Elephant Research and Conservation Centre, A Division of Asian Nature Conservation Foundation, Bangalore, India, 2p.

Ariffin, N., Yusof, N.A., Abdullah, J., Abd Rahman, S.F., Ahmad Raston, N.H., Kusnin, N. and Suraiya, S., 2020. Lateral flow immunoassay for naked eye detection of Mycobacterium tuberculosis. J. Sensors: 1-10.

Greenwald, R., Lyashchenko, O., Esfandiari, J., Miller, M., Mikota, S., Olsen, J.H., Ball, R., Dumonceaux, G., Schmitt, D., Moller, T., Payeur, J.B., Harris, B., Sofranko, D., Waters, W.R. and Lyashchenko, K.P. 2009. Highly accurate antibody assays for early and rapid detection of tuberculosis in African and Asian elephants. Clin. Vaccine Immunol. 16: 605-612.

Ishikawa, S., Ozeki, Y., Suga, S., Mukai, Y., Kobayashi, H., Inouchi, E., Kaboso, S.A., Gebretsadik, G., Dewi, D.N.S.S., Nishiyama, A., Tateishi, Y., Takihara, H., Okuda, S., Yoshida, S., Misawa, N. and Matsumoto, S. 2022. Monitoring IgG against Mycobacterium tuberculosis proteins in an Asian elephant cured of tuberculosis that developed from longterm latency. Sci. Rep. 12: 4310.

Lyashchenko, K.P., Greenwald, R., Esfandiari, J., Mikota, S., Miller, M., Moller, T., Vogelnest, L., Gairhe, K.P., RobbeAusterman, S., Gai, J. and Waters, W.R. 2012. Field application of serodiagnostics to identify elephants with tuberculosis prior to case confirmation by culture. Clin. Vaccine. Immunol. 19: 1269-1275.

Lyashchenko, K.P., Greenwald, R., Esfandiari, J., Olsen, J.H., Ball, R., Dumonceaux, G., Dunker, F., Buckley, C., Richard, M., Murray, S., Payeur, J.B., Andersen, P., Pollock, J.M., Mikota, S., Miller, M., Sofranko, D. and Waters, W.R. 2006. Tuberculosis in elephants: antibody responses to defined antigens of Mycobacterium tuberculosis, potential for early diagnosis, and monitoring of treatment. Clin. Vaccine. Immunol. 13: 722-732.

Mikota, S.K., Lyashchenko, K.P., Lowenstine, L., Agnew, D. and Maslow, J.N. 2015. Mycobacterial infections in elephants, In: Mukundan, H., Chambers, M.A., Waters, W.R., Larsen, M.H. (ed.) Tuberculosis, Leprosy and Mycobacterial Diseases of Man and Animals: The Many Hosts of Mycobacteria. pp. 259-276.

Payeur, J.B., Jarnagin, J.L., Marquardt, J.G. and Whipple, D.L. 2002. Mycobacterial isolations in captive elephants in the United States. Ann. N. Y. Acad. Sci. 969: 256-258.

Singh, A., Gupta, A.K., Gopinath, K., Sharma, P. and Singh, S. 2017. Evaluation of 5 Novel protein biomarkers for the rapid diagnosis of pulmonary and extrapulmonary tuberculosis: preliminary results. Sci. Rep. 7: 44121.

Cite this article

Abraham, D., Ravishankar, C., Deepa, P.M., Sumod, K., Bipin, K.C, Rajani, C.V. and Vijayakumar K. 2024. Evaluation of a Dot ELISA for tuberculosis screening in Asian elephants (Elephas maximus). J. Vet. Anim. Sci. 55(2):465-470

DOI: https://doi.org/10.51966/jvas.2024.55.2.465-470

Views
10
Downloads
5
Citations