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Volume 27 Issue 5
October 2012

    Shampur Narayan Madhusudana, Sundaramurthy Subha, Ullas Thankappan and Yajaman Belludi Ashwin. Evaluation of a Direct Rapid Immunohistochemical Test (dRIT) for Rapid Diagnosis of Rabies in Animals and Humans[J]. Virologica Sinica, 2012, 27(5): 299-302. doi: 10.1007/s12250-012-3265-6
    Citation: Shampur Narayan Madhusudana, Sundaramurthy Subha, Ullas Thankappan, Yajaman Belludi Ashwin. Evaluation of a Direct Rapid Immunohistochemical Test (dRIT) for Rapid Diagnosis of Rabies in Animals and Humans .VIROLOGICA SINICA, 2012, 27(5) : 299-302.  http://dx.doi.org/10.1007/s12250-012-3265-6
    shu

    Evaluation of a Direct Rapid Immunohistochemical Test (dRIT) for Rapid Diagnosis of Rabies in Animals and Humans

    cstr: 32224.14.s12250-012-3265-6

    • Department of Neurovirology, National Institute of Mental health and Neurosciences NIMHANS, Bangalore 560029, India

    • 通讯作者： Shampur Narayan Madhusudana, mshampur@hotmail.com; 
    • 收稿日期： 2012-06-14
      录用日期： 2012-09-07

    • Presently the gold standard diagnostic technique for rabies is the direct immunofluorescence assay ( dFA) which is very expensive and requires a high level of expertise. There is a need for more economical and user friendly tests, particularly for use in developing countries. We have established one such test called the direct rapid immunohistochemical test (dRIT) for diagnosis of rabies using brain tissue. The test is based on capture of rabies nucleoprotein (N) antigen in brain smears using a cocktail of biotinylated monoclonal antibodies specific for the N protein and color development by streptavidin peroxidase-amino ethyl carbazole and counter staining with haematoxollin. The test was done in parallel with standard FAT dFA using 400 brain samples from different animals and humans. The rabies virus N protein appears under light microscope as reddish brown particles against a light blue background. There was 100 % correlation between the results obtained by the two tests. Also, interpretation of results by dRIT was easier and only required a light microscope. To conclude, this newly developed dRIT technique promises to be a simple, cost effective diagnostic tool for rabies and will have applicability in field conditions prevalent in developing countries.

    Evaluation of a Direct Rapid Immunohistochemical Test (dRIT) for Rapid Diagnosis of Rabies in Animals and Humans

    • Department of Neurovirology, National Institute of Mental health and Neurosciences (NIMHANS), Bangalore 560029, India

    • Corresponding author: Shampur Narayan Madhusudana, mshampur@hotmail.com
    • Received Date: 14 June 2012
      Accepted Date: 07 September 2012
    • Presently the gold standard diagnostic technique for rabies is the direct immunofluorescence assay ( dFA) which is very expensive and requires a high level of expertise. There is a need for more economical and user friendly tests, particularly for use in developing countries. We have established one such test called the direct rapid immunohistochemical test (dRIT) for diagnosis of rabies using brain tissue. The test is based on capture of rabies nucleoprotein (N) antigen in brain smears using a cocktail of biotinylated monoclonal antibodies specific for the N protein and color development by streptavidin peroxidase-amino ethyl carbazole and counter staining with haematoxollin. The test was done in parallel with standard FAT dFA using 400 brain samples from different animals and humans. The rabies virus N protein appears under light microscope as reddish brown particles against a light blue background. There was 100 % correlation between the results obtained by the two tests. Also, interpretation of results by dRIT was easier and only required a light microscope. To conclude, this newly developed dRIT technique promises to be a simple, cost effective diagnostic tool for rabies and will have applicability in field conditions prevalent in developing countries.
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      1. Dean D J , Abelesth M K, Atanasiu P. 1996. The Fluorescent antibody test. In: Laboratory techniques in rabies, 4th ed. Meslin F X, Koprowsky H, Kaplan M M, eds. Geneva: World Health Organization. pp88-95.

      2. Durr S, Naissengar S, Mindekem R, et al. 2008. Rabies diagnosis for developing countries. PLoS Negl Trop Dis, 2:e206.

      3. Lembo T, Niezgoda M, Velasco-Villa A, et al. 2006. Evaluation of a direct, rapid immunohistochemical test for rabies diagnosis. Emerg Infect Dis, 12:310-313.

      4. Menezes R. 2008. Rabies in India. CMAJ, 178:564-566.
          doi: 10.1503/cmaj.071488

      5. Nagarajan T, Mohanasubramanian B, Seshagiri E V, et al. 2006. Molecular epidemiology of rabies virus isolates in India. J Clin Microbiol, 44:3218-3224.
          doi: 10.1128/JCM.00801-06

      6. Perrin P, Rollin P E, Sureau P. 1986. A rapid rabies enzyme immuno-diagnosis (RREID): a useful and simple technique for the routine diagnosis of rabies. J Biol Stand, 14:217-222.
          doi: 10.1016/0092-1157(86)90006-5

      7. Reddy G B, Singh R, Singh R P, et al. 2011. Molecular characterization of Indian rabies virus isolates by partial sequencing of nucleoprotein (N) and phosphoprotein (P) genes. Virus Genes, 43:13-17.
          doi: 10.1007/s11262-011-0601-0

      8. Saturday G A, King R, Fuhrmann L. 2009. Validation and operational application of a rapid method for rabies antigen detection. US Army Med Dep J, Jan-March, 42-45.

      9. Sudarshan M K, Madhusudana S N, Mahendra B J, et al. 2007. Assessing the burden of human rabies in India: results of a national multi-center epidemiological survey. Int J Infect Dis, 11:29-35.
          doi: 10.1016/j.ijid.2005.10.007

      10. Tao X Y, Niezgoda M, Du J L, et al. 2008. The primary application of direct rapid immunohistochemical test to rabies diagnosis in China. Chin J Exp Clin Virol, 22(3):168-170. (in Chinese)

      11. World Health Organization. 2005. Expert consultation on rabies, 1st report. WHO Tech Rep Ser 935, Geneva: WHO.

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      Evaluation of a Direct Rapid Immunohistochemical Test (dRIT) for Rapid Diagnosis of Rabies in Animals and Humans

        Corresponding author: Shampur Narayan Madhusudana, mshampur@hotmail.com
      • Department of Neurovirology, National Institute of Mental health and Neurosciences (NIMHANS), Bangalore 560029, India
      • Received Date: 14 June 2012
      • Accepted Date: 07 September 2012

      Abstract: Presently the gold standard diagnostic technique for rabies is the direct immunofluorescence assay ( dFA) which is very expensive and requires a high level of expertise. There is a need for more economical and user friendly tests, particularly for use in developing countries. We have established one such test called the direct rapid immunohistochemical test (dRIT) for diagnosis of rabies using brain tissue. The test is based on capture of rabies nucleoprotein (N) antigen in brain smears using a cocktail of biotinylated monoclonal antibodies specific for the N protein and color development by streptavidin peroxidase-amino ethyl carbazole and counter staining with haematoxollin. The test was done in parallel with standard FAT dFA using 400 brain samples from different animals and humans. The rabies virus N protein appears under light microscope as reddish brown particles against a light blue background. There was 100 % correlation between the results obtained by the two tests. Also, interpretation of results by dRIT was easier and only required a light microscope. To conclude, this newly developed dRIT technique promises to be a simple, cost effective diagnostic tool for rabies and will have applicability in field conditions prevalent in developing countries.

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