Production and evaluation of an antiserum for immunohistochemical detection of rabies virus in aldehyde fixed brain tissues
Keywords:
rabies, rabies virus, rabies vaccines, antibodies, fixatives, tissue fixation, immuno-histochemistry
Abstract
Introduction. The standard procedure for rabies diagnosis requires fresh samples of infected brain to be analyzed by two techniques, direct immunofluorescence and inoculation in mice. Rabies-infected, aldehyde-fixed brain tissues can be examined by immunohistochemistry, but the required commercial antibodies are scarce and expensive.Objectives. An anti-rabies antiserum was produced and tested to evaluate the effectiveness of rabies antigen detection in aldehyde preserved brain tissue.
Materials and methods. Rabbits were inoculated with a rabies vaccine produced in Vero cells (origin-African green monkey kidney). Anti-rabies antiserum was obtained and tested by immunohistochemistry in aldehyde-fixed brain sections of rabies-infected mice. Several experimental conditions were assayed. The usefulness of the antiserum in human pathology samples was also tested.
Results. The specificity of the antiserum was demonstrated for immunohistochemical detection of rabies antigen in fixed aldehydes nervous tissue both from experimental material and pathology archival collection. In addition, the antiserum was successful in detecting rabies virus under conditions that have been considered unfavorable for the preservation of antigens.
Conclusions. The inoculation of rabies vaccine in rabbits is an easy and safe procedure for obtaining antiserum useful for the detection of rabies antigen in samples of nervous tissue. Sections obtained on vibratome better preserve the viral antigenicity in comparison with paraffin-embedded tissues. This methods permit less expensive and more rapid immunohistochemical diagnosis of rabies.
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References
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14. Rodríguez G. Microscopía electrónica de la infección viral. Bogotá: Instituto Nacional de Salud; 1983. p. 119-39.
15. Rodríguez G, Sarmiento L. Rabia: el cuerpo de Negri. Biomédica. 1999;19:196-7.
16. Özkan Ö, Aylan O, Ates C, Celebi B. Production of anti-rabies immune sera. Etlik Veteriner Mikrobiyoloji Dergisi. 2004;15:49-54.
17. Redwan el-RM, Fahmy A, El Hanafy A, Abd El-Baky N, Sallam SM. Ovine anti-rabies antibody production and evaluation. Comp Immunol Microbiol Infect Dis. 2009;32:9-19.
18. Reblet C. Uso de las técnicas inmunocitoquímicas en neurobiología. En: Delgado JM, Ferrús A, Mora F, Rubia F, editores. Manual de neurociencia. Madrid: Editorial Síntesis; 1998. p. 387-8.
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2. Páez A, Rey G, Dulce A, Parra E, Díaz-Granados H. Brote de rabia humana transmitida por caninos en el distrito de Santa Marta, Colombia, 2006-2008. Biomédica. 2009;29:424-36.
3. Valderrama J, García I, Figueroa G, Rico E, Sanabria J, Rocha N, et al. Brotes de rabia humana transmitida por vampiros en los municipios de Bajo y Alto Baudó, departamento del Chocó, Colombia 2004-2005. Biomédica. 2006;26:387-96.
4. Kristensson K, Dastur DK, Manghani DK, Tsiang H, Bentivoglio M. Rabies: interactions between neurons and viruses. A review of Negri inclusion bodies. Neuropathol Appl Neurobiol. 1996;22:179-87.
5. Fu ZF, Jackson AC. Neuronal dysfunction and death in rabies virus infection. J Neurovirol. 2005;11:101-6.
6. Torres-Fernández O, Yepes GE, Gómez JE, Pimienta H. Efecto de la infección por el virus de la rabia sobre la expresión de parvoalbúmina, calbindina y calretinina en la corteza cerebral de ratones. Biomédica. 2004;24:63-78.
7. Torres-Fernández O, Yepes GE, Gómez JE, Pimienta H. Calbindin distribution in cortical and subcortical brain structures of normal and rabies infected mice. Int J Neurosci. 2005;115:1375-82.
8. Rengifo AC, Torres-Fernández O. Disminución del número de neuronas que expresan GABA en la corteza cerebral de ratones infectados por rabia. Biomédica. 2007;27:548-58.
9. Torres-Fernández O, Yepes GE, Gómez JE. Alteraciones de la morfología dendrítica neuronal en la corteza cerebral de ratones infectados con rabia: un estudio con la técnica de Golgi. Biomédica. 2007;27:605-13.
10. Lamprea N, Torres-Fernández O. Evaluación inmuno-histoquímica de la expresión de calbindina en el cerebro de ratones en diferentes tiempos después de la inoculación con el virus de la rabia. Colom Med. 2008;39(Suppl.3):7-13.
11. Trimarchi CV. Diagnostic evaluation. In: Jackson AC, Wunner WH, editores. Rabies. San Diego: Academic Press; 2002. p. 307-49.
12. Iwasaki Y, Tobita M. Pathology. In: Jackson AC, Wunner WH, editores. Rabies. San Diego: Academic Press; 2002. p. 283-306.
13. Sarmiento L, Rodríguez G, De Serna C, Boshell J, Orozco L. Detection of rabies virus antigens in tissue: immunoenzymatic method. Patología. 1999;37:7-10.
14. Rodríguez G. Microscopía electrónica de la infección viral. Bogotá: Instituto Nacional de Salud; 1983. p. 119-39.
15. Rodríguez G, Sarmiento L. Rabia: el cuerpo de Negri. Biomédica. 1999;19:196-7.
16. Özkan Ö, Aylan O, Ates C, Celebi B. Production of anti-rabies immune sera. Etlik Veteriner Mikrobiyoloji Dergisi. 2004;15:49-54.
17. Redwan el-RM, Fahmy A, El Hanafy A, Abd El-Baky N, Sallam SM. Ovine anti-rabies antibody production and evaluation. Comp Immunol Microbiol Infect Dis. 2009;32:9-19.
18. Reblet C. Uso de las técnicas inmunocitoquímicas en neurobiología. En: Delgado JM, Ferrús A, Mora F, Rubia F, editores. Manual de neurociencia. Madrid: Editorial Síntesis; 1998. p. 387-8.
19. Hockfield S, Carlson S, Evans C, Levitt P, Pintar J, Silberstein L. Selected methods for antibody and nucleic acid probes. New York: Cold Spring Harbor Laboratory Press; 1993. p. 111-226.
20. Castellanos JE, Guayacán OL, Castañeda DR, Hurtado H. Uso de una técnica de inmunoperoxidasa para la detección de virus de rabia en cortes gruesos de cerebro. Biomédica. 1998;18:141-6.
21. Ribeiro-Da-Silva A, Priestley J, Cuello C. Pre-embedding ultrastructural immunocytochemistry. En: Cuello E, editor. Immunohistochemistry II. Chichester: John Wiley & Sons; 1993. p. 181-227.
How to Cite
1.
Lamprea NP, Ortega LM, Santamaría G, Sarmiento L, Torres-Fernández O. Production and evaluation of an antiserum for immunohistochemical detection of rabies virus in aldehyde fixed brain tissues. biomedica [Internet]. 2010 Mar. 1 [cited 2024 May 18];30(1):146-51. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/162
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