FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Zoonoses Public Health / Detection of rabies virus antigen by the indirect rapid immu...
Zoonoses and public health2020; 67(6); 651-657; doi: 10.1111/zph.12745

Detection of rabies virus antigen by the indirect rapid immunohistochemistry test in equines and comparisons with other diagnostic techniques.

Abstract: Laboratory diagnosis of rabies in equines is essential for distinguishing the disease from other sources of encephalitis. Diagnosis by conventional techniques such as a direct fluorescent antibody test (dFAT) or viral isolation in mice or cell culture can be difficult, and the application of molecular biological methods may be necessary. We performed an indirect rapid immunohistochemistry test (iRIT) for the detection of the rabies virus (RABV) antigen in the central nervous system (CNS) of equines and compared the results with those of other diagnostic techniques. We reviewed result records from the Rabies Diagnosis Laboratory at Instituto Pasteur, São Paulo, Brazil, of 174 samples of equine CNS from July 2014 to June 2016, which were investigated by dFAT, rabies tissue culture infection test (RTCIT), mouse inoculation test (MIT) and reverse transcription-polymerase chain reaction (RT-PCR) followed by genetic sequencing. These samples, 29 presented divergent results among techniques and were selected for the performed in the iRIT. The detected positivity rate was 4/29 (14%) by dFAT, 5/28 (18%) by RTCIT, 10/29 (35%) by MIT and 26/27 (96%) by RT-PCR. We analysed 29 samples through imprints of the cortex, hippocampus, cerebellum and brainstem in slides fixed in 10% buffered formaldehyde. Eighteen samples were identified as positive (62%) by iRIT assay, representing a greater number of positive cases than that detected by dFAT, MIT and RTCIT but not by RT-PCR. Among the brain regions, the brainstem presented the highest positivity (78%), followed by the hippocampus (69%), cerebellum (67%) and cortex (67%). Our results provide evidence that iRIT can contribute to a rapid diagnosis of rabies in equines and that complementary tests should be used to improve diagnostic accuracy in this species.
© 2020 Blackwell Verlag GmbH.
Get Full Text
Publication Date: 2020-06-14 PubMed ID: 32537888DOI: 10.1111/zph.12745Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article focuses on the implementation of an indirect rapid immunohistochemistry test (iRIT) for detecting rabies virus in the central nervous system (CNS) of equines (horses). It is compared with other diagnostic techniques, ultimately concluding that iRIT can effectively contribute to quicker and more accurate rabies diagnosis in these animals.

Methodology and Sample Selection

  • The researchers examined the results from the Rabies Diagnosis Laboratory at the Instituto Pasteur in São Paulo, Brazil.
  • A total of 174 CNS samples from equines were reviewed, collected between July 2014 to June 2016.
  • These samples had previously been tested by other diagnostic techniques such as direct fluorescent antibody test (dFAT), rabies tissue culture infection test (RTCIT), mouse inoculation test (MIT) and reverse transcription-polymerase chain reaction (RT-PCR), followed by genetic sequencing.
  • Amongst these 174 samples, 29 showed divergent results among the techniques, and therefore chosen for further investigation with the iRIT

Analysis and Findings

  • The researchers analyzed imprints of the cortex, hippocampus, cerebellum, and brainstem, all significant parts of the CNS in equines, which were fixed on slides in 10% buffered formaldehyde.
  • The positivity rate detected by previously used methods varied: it was 14% by dFAT, 18% through RTCIT, 35% through MIT, and a high 96% with RT-PCR.
  • When these 29 samples were processed through iRIT, 18 samples (62%) were identified as positive for the rabies virus.
  • This represented a larger number of positive cases than detected by dFAT, MIT, and RTCIT, but not by RT-PCR.
  • Within the brain regions examined, the brainstem showed the highest positivity rate at 78%, followed by the hippocampus (69%), cerebellum (67%), and cortex (67%).

Conclusion

  • The study concludes that the iRIT can contribute significantly to faster and more accurate diagnosis of rabies in equines.
  • Furthermore, it underscores the value of using complementary tests for improving diagnostic accuracy in detecting rabies in this species.

Cite This Article

APA
Torquato RBC, Iamamoto K, Fernandes ER, Achkar S, Silva SR, Katz ISS, Guedes F. (2020). Detection of rabies virus antigen by the indirect rapid immunohistochemistry test in equines and comparisons with other diagnostic techniques. Zoonoses Public Health, 67(6), 651-657. https://doi.org/10.1111/zph.12745

Publication

Zoonoses and public health
ISSN: 1863-2378
NlmUniqueID: 101300786
Country: Germany
Language: English
Volume: 67
Issue: 6
Pages: 651-657

Researcher Affiliations

Torquato, Ranieli B C
  • Instituto Pasteur, São Paulo, Brazil.
Iamamoto, Keila
  • Instituto Pasteur, São Paulo, Brazil.
Fernandes, Elaine R
  • Instituto Pasteur, São Paulo, Brazil.
Achkar, Samira
  • Instituto Pasteur, São Paulo, Brazil.
Silva, Sandriana R
  • Instituto Pasteur, São Paulo, Brazil.
Katz, Iana S S
  • Instituto Pasteur, São Paulo, Brazil.
Guedes, Fernanda
  • Instituto Pasteur, São Paulo, Brazil.

MeSH Terms

  • Animals
  • Antigens, Viral / isolation & purification
  • Central Nervous System / virology
  • Horse Diseases / diagnosis
  • Horse Diseases / virology
  • Horses
  • Immunohistochemistry / methods
  • Immunohistochemistry / veterinary
  • Neurons / virology
  • Rabies / diagnosis
  • Rabies / veterinary
  • Rabies virus / immunology
  • Rabies virus / isolation & purification

References

This article includes 40 references
  1. Bourhy H, Rollin PE, Vincent J, Sureau P. Comparative field evaluation of the fluorescent-antibody test, virus isolation from tissue culture, and enzyme immunodiagnosis for rapid laboratory diagnosis of rabies. Journal of Clinical Microbiology 27(3), 519-523.
  2. Carrieri ML, Peixoto ZMP, Paciência MLB, Kotait I, Germano PML. Laboratory diagnosis of equine rabies and its implications for human postexposure prophylaxis. Journal of Virological Methods 138(1-2), 1-9.
    doi: 10.1016/j.jviromet.2006.07.005google scholar: lookup
  3. Coertse J, Weyer J, Nel LH, Markotter W. Improved PCR methods for detection of African rabies and rabies-related Lyssaviruses. Journal of Clinical Microbiology 48(11), 3949-3955.
    doi: 10.1128/jcm.01256-10google scholar: lookup
  4. Coetzer A, Sabeta CT, Markotter W, Rupprecht CE, Nel LH. Comparison of biotinylated monoclonal and polyclonal antibodies in an evaluation of a direct rapid immunohistochemical test for the routine diagnosis of rabies in Southern Africa. PLoS Neglected Tropical Diseases 8(9), e3189.
    doi: 10.1371/journal.pntd.0003189google scholar: lookup
  5. Dürr S, Naïssengar S, Mindekem R, Diguimbye C, Niezgoda M, Kuzmin I, Zinsstag J. Rabies diagnosis for developing countries. PLoS Neglected Tropical Diseases 2(3), e206.
    doi: 10.1371/journal.pntd.0000206google scholar: lookup
  6. Dyer JL, Niezgoda M, Orciari LA, Yager PA, Ellison JA, Rupprecht CE. Evaluation of an indirect rapid immunohistochemistry test for the differentiation of rabies virus variants. Journal of Virological Methods 190(1-2), 29-33.
    doi: 10.1016/j.jviromet.2013.03.009google scholar: lookup
  7. Fernandes ER, de Andrade HF, Lancellotti CLP, Quaresma JAS, Demachki S, da Costa Vasconcelos PF, Duarte MIS. In situ apoptosis of adaptive immune cells and the cellular escape of rabies virus in CNS from patients with human rabies transmitted by Desmodus rotundus. Virus Research 156(1-2), 121-126.
    doi: 10.1016/j.virusres.2011.01.006google scholar: lookup
  8. Fooks AR, Johnson N, Freuling CM, Wakeley PR, Banyard AC, McElhinney LM, Müller T. Emerging technologies for the detection of rabies virus: Challenges and hopes in the 21st century. PLoS Neglected Tropical Diseases 3(9), e530.
    doi: 10.1371/journal.pntd.0000530google scholar: lookup
  9. Green SL, Smith LL, Vernau W, Beacock SM. Rabies in horses: 21 cases (1970-1990). Journal of the American Veterinary Medical Association 200(8), 1133-1137.
  10. Keane DP, Little PB. Equine viral encephalomyelitis in Canada: A review of known and potential causes. Canadian Veterinary Journal 28(8), 497-504.
  11. Lembo T, Niezgoda M, Velasco-Villa A, Cleaveland S, Ernest E, Rupprecht CE. Evaluation of a direct, rapid immunohistochemical test for rabies. Emerging Infectious Diseases 12(2), 310-313.
    doi: 10.3201/eid1202.050812google scholar: lookup
  12. Lépine P. Laboratory techniques in rabies: Fermi-type vaccine. Monograph Series World Health Organization (23), 199-200.
  13. Lépine P, Atanasiu P. Production of antirabies serum of animal origin. Monograph Series World Health Organization (23), 299-303.
  14. Lima EF, Riet-Correa F, de Castro RS, Gomes AAB, Lima FS. Clinical signs, distribution of the lesions in the central nervous system and epidemiology of rabies in northeastern. Pesquisa Veterinária Brasileira 25(4), 250-264.
    doi: 10.1590/s0100-736x2005000400011google scholar: lookup
  15. Madhusudana SN, Subha S, Thankappan U, Ashwin YB. Evaluation of a direct rapid immunohistochemical test (dRIT) for rapid diagnosis of rabies in animals and humans. Virologica Sinica 27(5), 299-302.
    doi: 10.1007/s12250-012-3265-6google scholar: lookup
  16. Mani RS, Madhusudana SN. Laboratory diagnosis of human rabies: Recent advances. Scientific World Journal 2013, 1-10.
    doi: 10.1155/2013/569712google scholar: lookup
  17. Middel K, Fehlner-Gardiner C, Pulham N, Buchanan T. Incorporating direct rapid immunohistochemical testing into large-scale wildlife rabies surveillance. Tropical Medicine and Infectious Disease 2(3), 21.
    doi: 10.3390/tropicalmed2030021google scholar: lookup
  18. Niezgoda M, Rupprecht CE. Standard operating procedure for the direct rapid immunohistochemistry test for the detection of rabies virus antigen. National Laboratory Training Network Course (1-16).
  19. Patrick EM, Bjorklund BM, Kirby JD, Nelson KM, Chipman RB, Rupprecht CE. Enhanced rabies surveillance using a direct rapid immunohistochemical test. Journal of Visualized Experiments: Jove 146, e59416.
    doi: 10.3791/59416google scholar: lookup
  20. Peixoto ZMP, Cunha EMS, Sacramento DRV, Souza MCAM, Silva LHQD, Germano PL, Kotait I. Rabies laboratory diagnosis: Peculiar features of samples from equine origin. Brazilian Journal of Microbiology 31, 72-75.
    doi: 10.1590/s1517-83822000000100017google scholar: lookup
  21. Prabhu K, Isloor S, Veeresh B, Rathnamma D, Sharada R, Das L, Rahman S. Application and comparative evaluation of fluorescent antibody, immunohistochemistry and reverse transcription polymerase chain reaction tests for the detection of rabies virus antigen or nucleic acid in brain samples of animals suspected of rabies in India. Veterinary Sciences 5(1), 24.
    doi: 10.3390/vetsci5010024google scholar: lookup
  22. Rahmadane I, Certoma AF, Peck GR, Fitria Y, Payne J, Colling A, Allen JD. Development and validation of an immunoperoxidase antigen detection test for improved diagnosis of rabies in Indonesia. PLoS Neglected Tropical Diseases 11(11), e0006079.
    doi: 10.1371/journal.pntd.0006079google scholar: lookup
  23. Robardet E, Andrieu S, Rasmussen TB, Dobrostana M, Horton DL, Hostnik P, Cliquet F. Comparative assay of fluorescent antibody test results among twelve European National Reference Laboratories using various anti-rabies conjugates. Journal of Virological Methods 191, 88-94.
    doi: 10.1016/j.jviromet.2013.03.027google scholar: lookup
  24. Robardet E, Picard-Meyer E, Andrieu S, Servat A, Cliquet F. International interlaboratory trials on rabies diagnosis: An overview of results and variation in reference diagnosis techniques (fluorescent antibody test, rabies tissue culture infection test, mouse inoculation test) and molecular biology techniques. Journal of Virological Methods 177, 15-25.
    doi: 10.1016/j.jviromet.2011.06.004google scholar: lookup
  25. Rudd RJ, Trimarchi CV. Development and evaluation of an in vitro virus isolation procedure as a replacement for the mouse inoculation test in rabies diagnosis. Journal of Clinical Microbiology 27(11), 2522-2528.
    doi: 10.1128/jcm.27.11.2522-2528.1989google scholar: lookup
  26. Rupprecht CE, Cliquet F, Fehlner-Gardiner C, Fooks AR, Mueller T, Sabeta C, Slate D. Progress in the development of a direct rapid immunohistochemical test for diagnosing rabies. OIE Bulletin 3(87), 95.
  27. Rupprecht CE, Xiang Z, Servat A, Franka R, Kirby J, Ertl HCJ. Additional progress in the development and application of a direct, rapid immunohistochemical test for rabies diagnosis. Veterinary Sciences 5(2), 59.
    doi: 10.3390/vetsci5020059google scholar: lookup
  28. Secretaria de Vigilância Sanitária do Ministério da Saúde (2019). Retrieved from http://www.saude.gov.br/saude-de-a-z/raiva#epidemiologia
  29. Singh R, Singh KP, Cherian S, Saminathan M, Kapoor S, Reddy GBM, Dhama K. Rabies - epidemiology, pathogenesis, public health concerns and advances in diagnosis and control: A comprehensive review. The Veterinary Quarterly 37(1), 212-251.
    doi: 10.1080/01652176.2017.1343516google scholar: lookup
  30. Smith LL, Clare DA. A clinical note on equine rabies. Canadian Veterinary Journal 13(8), 193.
  31. Stein LT, Rech RR, Harrison L, Brown CC. Immunohistochemical study of rabies virus within the central nervous system of domestic and wildlife species. Veterinary Pathology 47(4), 630-636.
    doi: 10.1177/0300985810370013google scholar: lookup
  32. Summers BA, Cummings JF, De Lahunta A. Inflammatory diseases of the central nervous system. Veterinary Neuropathology p. 95-188.
  33. Tabel H, Charlton KM. The diagnosis of rabies in a horse by brain neutralization test. Canadian Journal of Comparative Medicine 38, 345-346.
  34. Tierkel ES. Rapid microscopic examination for Negri bodies and preparation of specimens for biological test. pp. 41-55.
  35. Trimarchi CV, Smith JS. Diagnostic evaluation. pp. 308-349.
  36. WHO, Rupprecht CE, Fooks AR, Abela-Ridder B. Virus isolation in animals: the mouse inoculation. Laboratory techniques in rabies, Volume 1, Chapter 8 5th ed., p. 74-84.
  37. WHO, Rupprecht CE, Fooks AR, Abela-Ridder B. Virus isolation in cell culture: the rabies tissue culture infection test. Laboratory techniques in rabies, Volume 1, Chapter 9 5th ed., p. 85-95.
  38. WHO, Rupprecht CE, Fooks AR, Abela-Ridder B. The direct fluorescent antibody test. Laboratory techniques in rabies, Volume 1, Chapter 11 5th ed., p. 108-129.
  39. WHO, Rupprecht CE, Fooks AR, Abela-Ridder B. The direct rapid immunohistochemistry test for the detection of lyssavirus antigens. Laboratory techniques in rabies, Volume 1, Chapter 12 5th ed., p. 130-135.
  40. WHO, Rupprecht CE, Fooks AR, Abela-Ridder B. Conventional pan-lyssavirus reverse transcriptase polymerase chain reaction. Laboratory techniques in rabies, Volume 2, Chapter 27 5th ed., p. 1-16.
Subscribe to our newsletter
Join 99,383 horse owners like you who receive our email updates on horse health and nutrition.