Simplified fluorescent inhibition microtest for the titration of rabies neutralizing antibodies

Favoretto, S.R.; Carrieri, M.L.; Tino, M.S.; Zanetti, C.R.; Pereira, O.A.C.

doi:10.1590/S0036-46651993000200009

Abstracts

A simplified fluorescence inhibition microtest (SFIMT) was standardized for the evaluation of antirabies serum neutralizing antibodies based on the rapid fluorescent focus inhibition test (RFFIT) and the fluorescence inhibition microtest (FIMT). The simplified test showed reproductibility similar to that of the FIMT with advantages as easier executation and quicker reading. A simple pre-treatment of Brazilian microplates produced for immune enzymatic assays (PROSIL) gave equivalent results and substantial coast reduction, in relation to imported plates (DIFCO). The simplified test can be easily implemented in less sophisticated laboratories, as alternative to the mouse serum neutralization test, still the most largely employed in Brazil, or even to others as RFFIT and FIMT.

Simplified Fluorescence Inhibition Microtest; rabies antibodies

No presente trabalho foi padronizada uma microtécnica simplificada para a avaliação de anticorpos neutralizantes anti-rábicos com base no RFFIT e no FIMT. Este teste mostrou reprodutibilidade comparável a dos testes originais, sendo de execução mais simples e de leitura mais rápida. Uma microplaca de fabricação nacional para testes imuno-enzimáticos (PROSIL Ind. e Com.) pode ser utilizada com resultados equivalentes aos de microplacas Difco ou similar, com substancial redução de custos, mediante tratamento simples e rápido. O teste estudado pode ser implantado facilmente em laboratórios menos sofisticados substituindo com grandes vantagens a prova de soro-neutralização em camundongos ainda a mais empregada rotineiramente no Brasil.

DIAGNOSTIC TRIAL

Simplified fluorescent inhibition microtest for the titration of rabies neutralizing antibodies

Microteste de inibição de fluorescência simplificado para titulação de anticorpos antirábicos

S.R. Favoretto^I; M.L. Carrieri^I; M.S. Tino^II; C.R. Zanetti^I; O.A.C. Pereira^I

^IInstituto Pasteur - São Paulo. São Paulo, SP. Brasil

^IIEscola Paulista de Medicina - Disciplina de Imunologia. São Paulo, SP. Brasil

^{Address for correspondence} Address for correspondence: Silvana R. Favoretto Instituto Pasteur-São Paulo Avenida Paulista, 393 CEP 01311-000 - São Paulo - SP - Brasil

SUMMARY

A simplified fluorescence inhibition microtest (SFIMT) was standardized for the evaluation of antirabies serum neutralizing antibodies based on the rapid fluorescent focus inhibition test (RFFIT) and the fluorescence inhibition microtest (FIMT).

The simplified test showed reproductibility similar to that of the FIMT with advantages as easier executation and quicker reading.

A simple pre-treatment of Brazilian microplates produced for immune enzymatic assays (PROSIL) gave equivalent results and substantial coast reduction, in relation to imported plates (DIFCO).

The simplified test can be easily implemented in less sophisticated laboratories, as alternative to the mouse serum neutralization test, still the most largely employed in Brazil, or even to others as RFFIT and FIMT.

Key words: Simplified Fluorescence Inhibition Microtest; rabies antibodies

RESUMO

No presente trabalho foi padronizada uma microtécnica simplificada para a avaliação de anticorpos neutralizantes anti-rábicos com base no RFFIT e no FIMT.

Este teste mostrou reprodutibilidade comparável a dos testes originais, sendo de execução mais simples e de leitura mais rápida.

Uma microplaca de fabricação nacional para testes imuno-enzimáticos (PROSIL Ind. e Com.) pode ser utilizada com resultados equivalentes aos de microplacas Difco ou similar, com substancial redução de custos, mediante tratamento simples e rápido.

O teste estudado pode ser implantado facilmente em laboratórios menos sofisticados substituindo com grandes vantagens a prova de soro-neutralização em camundongos ainda a mais empregada rotineiramente no Brasil.

Full text available only in PDF format.

Texto completo disponível apenas em PDF.

ACKNOWLEDGEMENTS

The authors gratefully acknowledge Mrs. Dolores Ayako Yoda, Mrs. Maria Aparecida da Silva and Mr. José Luiz de Carvalho Jr. for the technical assistence. This work was supported by The Health Secretary of the State of São Paulo, CNPq and FINEP.

Recebido para publicação em 7/8/1992

Aceito para publicação em 3/3/1993.

1. DIAZ, A.M.; ARISPE, E.; BRUNEL, C.; CAVANDOLI, C.; DELLEPIANE, N. & MIRANDA, A. - La tecnica de contrainmunoelectroforesis para la determinación de anticuerpos antirrabicos. Bol. Ofic. sanit. panamer., 101: 255-261, 1986.
2. GOUGH, P.M. & DIERKS, R.E. - Passive hemagglutina-üon test for antibodies against rabies virus. Bull. Wld. Hlth. Org., 45: 741-745, 1971.
3. JOHNSON, H.N. & LEACH, C.N. - Comparative susceptibility of different strains of mice to rabies virus. Amer. J. Hyg., 32: 38-45, 1940.
4. LAFON, M.; BOURHY, H. & SUREAU, P. - Immunity against the European bat rabies (Duvenhage) virus induced by rabies vaccines: an experimental study in mice. Vaccine, 6: 362-368, 1988.
5. PEREIRA, O.A.C.; RAPHAELIAN, T. & COUTINHO, N. - Complement fixation test in evaluation of immunity against rabies. Rev. Microbiol. (Săo Paulo), 1: 85-91, 1970.
6. PERRIN, P.; PORTNOI, D. & SUREAU, P. - Étude de l'adsorption et de la pénétration du virus rabique: interaction avec les cellules BHK21 et des membranes artificielles. Ann. Virol. (Paris), 133: 403, 1982.
7. SMITH, J.S.; YAGER, P.A. & BAER, G.M. - A rapid reproducible test for determining rabies neutralizing antibody. Bull. Wld. Hlth. Org., 48: 535-541, 1973.
8. THOMAS, J.B.; SIKES, R.K. & RICKE, A.S. - Evaluation of indirect fluorescent antibody technique for detection of rabies antibody in human sera. J. Immunol., 91: 721-723, 1963.
9. WEBSTER, L.T. & DAWSON Jr., J.R. - Early diagnosis of rabies by mouse inoculation. Measurement of humoral immunity to rabies by mouse protection test. Proc. Soc. exp. Biol. (N.Y.), 32: 570-573, 1935.
10. ZALAN, E.; WILSON, C. & PUKITIS, D. - A microtest for quantitation of, rabies virus neutralizing antibodies. J. biol. Stand., 7: 213-220, 1979.
11. ZANETTI, C.R.; TINO, M.S.; CHAMELET, E.L.B.; ISHIZUKA, M.M. & PEREIRA, O.A.C. - Simplification of immune adherence hemagglulination test for detection of rabies antibodies in human serum. Rev. Inst. Med. trop. S. Paulo, 31: 341-345, 1989.

Address for correspondence:

Silvana R. Favoretto

Instituto Pasteur-São Paulo

Avenida Paulista, 393

CEP 01311-000 - São Paulo - SP - Brasil

Publication Dates

Publication in this collection
27 July 2006
Date of issue
Apr 1993

History

Accepted
03 Mar 1993
Received
07 Aug 1992

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. DIAZ, A.M.; ARISPE, E.; BRUNEL, C.; CAVANDOLI, C.; DELLEPIANE, N. & MIRANDA, A. - La tecnica de contrainmunoelectroforesis para la determinación de anticuerpos antirrabicos. Bol. Ofic. sanit. panamer., 101: 255-261, 1986.

[2] 2. GOUGH, P.M. & DIERKS, R.E. - Passive hemagglutina-üon test for antibodies against rabies virus. Bull. Wld. Hlth. Org., 45: 741-745, 1971.

[3] 3. JOHNSON, H.N. & LEACH, C.N. - Comparative susceptibility of different strains of mice to rabies virus. Amer. J. Hyg., 32: 38-45, 1940.

[4] 4. LAFON, M.; BOURHY, H. & SUREAU, P. - Immunity against the European bat rabies (Duvenhage) virus induced by rabies vaccines: an experimental study in mice. Vaccine, 6: 362-368, 1988.

[5] 5. PEREIRA, O.A.C.; RAPHAELIAN, T. & COUTINHO, N. - Complement fixation test in evaluation of immunity against rabies. Rev. Microbiol. (Săo Paulo), 1: 85-91, 1970.

[6] 6. PERRIN, P.; PORTNOI, D. & SUREAU, P. - Étude de l'adsorption et de la pénétration du virus rabique: interaction avec les cellules BHK21 et des membranes artificielles. Ann. Virol. (Paris), 133: 403, 1982.

[7] 7. SMITH, J.S.; YAGER, P.A. & BAER, G.M. - A rapid reproducible test for determining rabies neutralizing antibody. Bull. Wld. Hlth. Org., 48: 535-541, 1973.

[8] 8. THOMAS, J.B.; SIKES, R.K. & RICKE, A.S. - Evaluation of indirect fluorescent antibody technique for detection of rabies antibody in human sera. J. Immunol., 91: 721-723, 1963.

[9] 9. WEBSTER, L.T. & DAWSON Jr., J.R. - Early diagnosis of rabies by mouse inoculation. Measurement of humoral immunity to rabies by mouse protection test. Proc. Soc. exp. Biol. (N.Y.), 32: 570-573, 1935.

[10] 10. ZALAN, E.; WILSON, C. & PUKITIS, D. - A microtest for quantitation of, rabies virus neutralizing antibodies. J. biol. Stand., 7: 213-220, 1979.

[11] 11. ZANETTI, C.R.; TINO, M.S.; CHAMELET, E.L.B.; ISHIZUKA, M.M. & PEREIRA, O.A.C. - Simplification of immune adherence hemagglulination test for detection of rabies antibodies in human serum. Rev. Inst. Med. trop. S. Paulo, 31: 341-345, 1989.