Veterinary World

     Open access and peer reviewed journal  

ISSN (Online): 2231-0916


Home l Editorial board l Instructions for authors l Reviewer guideline l Open access policy l Archives l FAQ

Open Access

Research (Published online: 31-01-2016)

18. Comparison of rapid immunodiagnosis assay kit with molecular and immunopathological approaches for diagnosis of rabies in cattle - Ajaz Ahmad and C. K. Singh

Veterinary World, 9(1): 107-112



   doi: 10.14202/vetworld.2016.107-112



Ajaz Ahmad: Department of Veterinary Pathology, College of Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 004, Punjab, India;

C. K. Singh: Department of Veterinary Pathology, College of Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 004, Punjab, India;


Received: 09-10-2015, Revised: 20-12-2015, Accepted: 28-12-2015, Published online: 31-01-2016


Corresponding author: Ajaz Ahmad, e-mail:

Citation: Ahmad A, Singh CK (2016), Comparison of rapid immunodiagnosis assay kit with molecular and immunopathological approaches for diagnosis of rabies in cattle, Veterinary World 9(1): 107-112.

Aim: Presently, diagnosis of rabies is primarily based on, conventional fluorescent antibody technique (FAT), immunopathological and molecular techniques. Recently, rapid immunodiagnostic assay (RIDA) - A monoclonal antibodybased technique has been introduced for rapid diagnosis of rabies. The present investigation is envisaged to study the efficacy of RIDA kit for the diagnosis of rabies in cattle.

Materials and Methods: About 11 brain samples from cattle, clinically suspected for rabies, were screened by the FAT, Heminested reverse transcriptase polymerase chain reaction (HnRT-PCR), Immunohistochemistry (IHC), and RIDA.

Results: The sensitivity for detection of rabies from brain tissue by RIDA was 85.7% as compared to 100% by IHC as well as HnRT-PCR. The accuracy of detection of rabies by RIDA was 91.6% as compared to 100% that of IHC and HnRT-PCR, whereas specificity of RIDA was 100% like that of the IHC and HnRT-PCR.

Conclusion: Despite a comparatively low-sensitivity and accuracy of RIDA, latter can still be useful in screening a large number of field samples promptly. However, it is recommended that negative results with RIDA in cattle need to be authenticated by suitable alternative diagnostic approaches.

Keywords: cattle, diagnosis, fluorescent antibody technique, heminested reverse transcriptase, immunohistochemistry, rabies.

1. Lembo, T., Niezgoda, M., Velasco-Villa, A., Cleaveland, S., Ernest, E. and Rupprecht, C.E. (2006) Evaluation of a direct, rapid immunohistochemical test for rabies diagnosis. Emerg. Infect. Dis., 12: 310-313.
PMid:16494761 PMCid:PMC3294322
2. Sharma, P., Singh, C.K., Sood, N.K., Sandhu, B.S., Gupta, K. and Brar, A.P.S. (2014) Diagnosis of rabies from brain: Comparison of histochemical and histopathological approaches. Indian J. Vet. Pathol., 38(4): 269-272.
3. Wahan, S., Sandhu, B.S., Singh, C.K., Gupta, K., Sood, N.K. and Kaw, A. (2012) Comparison of clinico-pathological, immunohistochemical and immunofluorescent techniques for diagnosis of rabies in animals. Indian J. Anim. Sci., 82(1), 3-8. .
4. Sandhu, B.S., Sood, N.K., Awahan, S., Singh, C.K. and Gupta, K. (2011) Immunohistochemistry, histopathology, quantitative morphometry of Negri bodies in the brain of rabid animals. Indian J. Vet. Pathol., 35(2): 117-122.
5. David, D., Yakobson, B., Rotenberg, D., Dveres, N., Davidson, I. and Stram, Y. (2002) Rabies virus detection by RT-PCR in decomposed naturally infected brains. Vet Microbiol., 87: 111-118.
6. Yang, J., Hooper, D.C., Wunner, W.H., Koprowski, H. and Dietzschold, B.F. (1998) The specificity of rabies virus RNA encapsidation by nucleoprotein. Virology, 242: 107-117.
7. Pranoti, S., Singh, C.K. and Deepti, N. (2015) Comparison of immunochromatographic diagnostic test with heminested reverse transcriptase polymerase chain reaction for detection of rabies virus from brain samples of various species. Vet. World, 8(2): 135-138.
8. WHO. (1992) Expert Committee on Rabies. 8th Report. WHO Technical Report Series, No. 824. World Health Organization, Geneva. p7, 8.
9. Meslin, F.X., Kaplan, M.M. and Koprowski, H. (1996) Laboratory Diagnosis of Rabies. WHO, Geneva. p88-95.
10. Luna, L.G. (1968) Manual of Histological Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. The Blakiston Division, McGraw-Hill Book Co., New York.
11. Pranoti, S., Singh, C.K., Sood, N.K., Gupta, K., Sandhu, B.S. and Brar, A.P.S. (2015) Immunohistochemical detection of rabies in dogs from skin. Indian J. Canine Pract., 7(1): 37-40.
12. Pedroso, P.M.O., Pescador, C.A., Bandarra, P.M., Raymundo, D.L., Borba, M.R., Wouters, F., Bezerra-Junior, P.S. and Driemeier, D. (2008) Standardization of immunohistochemistry technique for detection of rabies virus in formalin-fixed and paraffin-embedded tissue samples from central nervous system of cattle. Pesqui. Vet. Bras. 28(12): 627.
13. Dong, K.Y., Eun, K.S., Yoon, I.O., Kyung, W.L., Chung, S.L., Seo, Y.K., Jeong, A.L. and Jae, Y.S. (2012) Comparison of four diagnostic methods for detecting rabies viruses circulating in Korea. J. Vet. Sci., 13(1): 43-48.
14. Dean, D.J., Abelseth, M.K. and Atanasiu, P. (1996) The fluorescent antibody test. In: Meslin, F.X., Kaplan, M.M. and Koprowski, H., editors. Laboratory Techniques in Rabies. 4th ed. WHO, Geneva. p88-95.
15. Ehizibolo, D.O., Nwosuh, C., Ehizibolo, E.E. and Kia, G.S.N. (2009) Comparison of the fluorescent antibody test and direct microscopic examination for rabies diagnosis at the National Veterinary Research Institute, Vom, Nigeria. Afr. J. Biomed. Res., 12: 1.
16. Singh, H. (1999) Experimental rabies in buffalo calves with street rabies virus. M.V.Sc. Thesis, Punjab Agricultural University, Ludhiana, India.
17. Sumedha, A. (2010) Anti-mortem and post-mortem detection of rabies virus antigen in natural cases of rabies in animals - An immunopathological study. M.V.Sc. Thesis. Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, India.
18. Last, R.D., Jardine, J.E., Smit, M.M.E. and Van Der Lugt, J.J. (1994) Application of immunoperoxidase techniques to formalin-fixed brain tissue for the diagnosis of rabies in southern Africa. J. Vet. Res., 61(2): 183-187.
19. Gunawardena, G.S.P. and Blakemore, W.F. (2007) Proceedings of the Peradeniya University Research Sessions, Sri Lanka. Vol. 12. No. 1.p168.
20. Rissi, D.R., Fighera, R.A., Irigoyen, L.F., Kommers, G.D. and Claudio, S.L.B. (2008) Occurrence of rabies in sheep in Rio Grande do Sul, Brazil. Pesqui. Vet. Bras., 28(10): 495.
21. Soares, R.M., Bernardi, F., Sakamoto, S.M., Heinemann, M.B., Cortez, A., Alves, L.M., Meyer, A.D., Ito, F.H. and Richtzenhain, L.J. (2002) A heminested polymerase chain reaction for the detection of Brazilian rabies isolates from vampire bats and herbivores. Mem. Inst. Oswaldo Cruz, 97: 109-111.
22. Orłowska, A., Smreczak, M., Trębas, P. and Żmudziński, J.F. (2008) Comparison of real-time PCR and heminested RT-PCR methods in the detection of rabies virus infection in bats and terrestrial animals. Bull. Vet. Inst. Pulawy, 52: 313-318.
23. Servat, A., Picard-Meyer, E., Robardet, E., Muzniece, Z., Must, K. and Cliquet, F. (2012) Evaluation of a rapid immunochromatographic diagnostic test for the detec-tion of rabies from brain material of European mammals. Biologicals, 40(1): 61-66.
24. Kang, B., Oh, J., Lee, C., Park, B.K., Park, Y., Hong, K., Lee, K., Cho, B. and Song, D. (2007) Evaluation of a rapid immunodiagnostic test kit for rabies virus. J. Virol. Methods, 145(1): 30-36.