SciELO - Scientific Electronic Library Online

 
vol.24 número2Comparação do parasitismo da veia central da supra-renal com o de outros tecidos em chagásicos crônicosAcidente crotálico na infância: aspectos clínicos, laboratoriais, epidemiológicos e abordagem terapêutica índice de autoresíndice de assuntospesquisa de artigos
Home Pagelista alfabética de periódicos  

Serviços Personalizados

Journal

Artigo

Indicadores

Links relacionados

Compartilhar


Revista da Sociedade Brasileira de Medicina Tropical

versão impressa ISSN 0037-8682

Rev. Soc. Bras. Med. Trop. vol.24 no.2 Uberaba abr./jun. 1991

http://dx.doi.org/10.1590/S0037-86821991000200003 

ARTICLES

 

Two competitive enzyme immunoassays for the detection of IgG class antibodies to hepatitis a antigen

 

 

Claudia Lamarca Vitral; Ana Maria Coimbra Gaspar; Clara Fumiko Tachibana Yoshida

Adress to correspondence

 

 


ABSTRACT

Two competitive enzyme immunoassays (EIA) techniques were developed: in the first (COMP-1), test sera were added together with HAV antigen on anti-HAV IgG-coated wells followed by an anti-HA VHRP conjugate; in the second (COMP-2), test sera and anti-HA VHRP conjugate competed for HAV epitopes previously adsorbed to anti-HA V IgG-coated wells. Both procedures used tetramethylbenzidine (TMB) as a substrate. Both competitive tests were shown to be reproducible and suitable for routine diagnosis and research purposes.

Keywords: Enzyme immunoassay. Anti-HAV. Hepatitis A.


RESUMO

Foram desenvolvidos dois ensaios imunoenzimáticos (EIA) competitivos: no primeiro (COMP-1) colocou-se numa placa sensibilizada com anti-HAVIgG as amostras teste juntamente como antígeno HA Vea seguir o conjugado anti-HA VHRP; no segundo (COMP-2), as amostras teste e o conjugado anti-HAV HRP competem pelos epitopos do antígeno HAV previamente absorvido na placa sensibilizada do anti-HAV IgG. O substrato utilizado foi tetrametilbenzidina (TMB). Ambas as técnicas mostraram ser produtíveis e aplicáveis para fins de diagnóstico e pesquisa.

Palavras-chave: Enzyme immmunoassay. Anti-HAV. Hepatitis A.


 

 

Full text available only in PDF format.

Texto completo disponível apenas em PDF.

 

 

REFERENCES

1. Bos E, Van Der Doelen AA, Van Rooy N, Schuurs AHWM. 3,3',5,5' - Tetramethylbenzidine as an ames test negative chromogen for horseradish peroxidase in enzyme-immunoassay. Journal of Immunoassay 2: 187-204, 1981.         [ Links ]

2. Camargo IF, Gaspar AMC, Yoshida CFT. Comparative ELISA reagents for detection of hepatitis B surface antigen (HBsAg). Memórias do Instituto Oswaldo Cruz 82: 181-187, 1987.         [ Links ]

3. Codd AA, Sprott MS, Narang HR, Crone PB, Turner RH. Competitive enzyme-linked immunosorbent assay for Treponema pallidum antibodies. Journal of Mfedical Microbiology 26: 153-157, 1988.         [ Links ]

4. Coulepis AG, Veale MF, MacGregor A, Komitschuk M. Detection of hepatitis A virus and antibody by solid- phase radioimmunoassay and enzyme-linked immunosorbent assay with monoclonal antibodies. Journal of Clinical Microbiology 22: 119-124, 1985.         [ Links ]

5. Daemer RJ, Feinstone SM, Gust ID, Purcell RH. Propagation of human hepatitis A virus in african green monkey Kidney cell culture: primary isolation and serial passage. Infection and Immunity 32: 388-393, 1981.         [ Links ]

6. Feinstone SM, Kapikian AZ, Purcell RH. Hepatitis A: detection by immune electron microscopy of a virus-like antigen associated with acute illness. Science 182:1026- 1028, 1973.         [ Links ]

7. Flehming B. Hepatitis A virus in cell culture: I. Propagation of different hepatitis A virus isolates in a fetal rhesus monkey kidney cell line (FRhK-4). Medical Microbiology and Immunology 168: 239-248, 1980.         [ Links ]

8. Flehming B. Hepatitis A in cell culture: II. Growth characteristics of hepatitis A virus in FRhK-4/R cells. Medical Microbiology and Immunology 170: 73-81, 1981.         [ Links ]

9. FlehmingB, Pfisterer M, VallbrachtA. Hepatitis A virus in cell culture: IV. Comparison of cell-cultured produced HAV with stool derived HAV in diagnostic test systems. Medical Microbiology and Immunology 172: 207-213, 1984.         [ Links ]

10. Frosner GG, Deinhardt F, Scheid R, Gauss-Muller V, Holmes N, Messelberger V, Siegel G, Alexander JJ. Propagation of human hepatitis A virus in a hepatoma cell line. Infection 7: 303-306, 1979.         [ Links ]

11. Gauss-Muller V, Frosner GG, Deinhardt F. Propagation of hepatitis A virus in human embryo fibroblasts. Journal of Medical Virology 7: 233-239, 1981.         [ Links ]

12. GirardetC, PeitrequinR, Frei PC. Serodiagnosis of viral hepatitis A rise in antibody titre and evaluation of three methods for detecting early and late antibodies. Experientia 39: 902-904, 1983.         [ Links ]

13. Gust ID, Lehmann NI, Crowe S, McCrowe M, Locamini SA, Lucas R. The origin of HM-175 strain of hepatitis A virus. The Journal of Infectious Diseases 151: 365-366 1985.         [ Links ]

14. Kojima H, Shibayama T, Sato A, Suzuki S, Ichida F, Hamada C. Propagation of human hepatitis A virus in conventional cell lines. Journal of Medical Virology 7: 272-286, 1981.         [ Links ]

15. Miller WJ, Provost PJ, McAleer WJ, Ittensohn OL, Villarejos VM, Hilleman MR. Specific immuneadhrence assay for human hepatitis A antibody. Application to diagnostic and epidemiologic investigation. Proceedings of the Society for Experimental Biology and Medicine 49: 254-261, 1975.         [ Links ]

16. Parry JV. Hepatitis A infection: guidelines for development of satisfactory assays for laboratory diagnosis. Medical Laboratory Sciences 38: 303-311, 1981.         [ Links ]

17. Provost PJ, Ittensohn OL, Villarejos VM, Hilleman MR. A specific complement - fixation test for human hepatitis A employing CR326 virus antigen. Diagnosis and epidemiology (38669). Proceedings of the Society for Experimental Biology and Medicine 148: 962-969, 1975.         [ Links ]

18. Provost PJ, Hilleman MR. Propagation of human hepatitis A virus in cell culture in vitro. Proceedings of the Society for Experimental Biology and Medicine 160: 213-221, 1979.         [ Links ]

19. Purcell RH, Wong DC, Alter HJ, Holland PV. Microtiter solid-phase radioimmunoassay for hepatitis B antigen. Applied Microbiology 26: 478-484, 1973.         [ Links ]

20. Purcell RH, Wong DC, Moritsugu Y, Dienstag JL, Routenberg JA, Boggs J. A microtiter solid-phase ra-dioimmunoassay for hepatitis A antigen and antibody. The Journal of Immunology 116: 349-356, 1976.         [ Links ]

21. Rakela J, Fay OH, Svevenson D, Gordon I, Mosley JW. Similarities of two hepatitis A virus strains. Bulletin of the World Health Organization 54: 561-564, 1976.         [ Links ]

22. Tedder RS, Mortimer PP, Bridget Lord R. Detection of antibody to varicella-zoster virus by competitive and IgM-antibody capture immunoassay. Journal of Medical Virology 8: 89-101, 1981.         [ Links ]

23. Wallace RE, Vasington PJ, Petricciani JC, Hopps HE, Lorenz DE, Kadanka Z. Development and characterization of cell lines from subhuman primates. in vitro 8: 333-341, 1973.         [ Links ]

 

 

Adress to correspondence:
Dra. Claudia Lamarca Vitral.
Depto. of Virology/FIOCRUZ.
Av. Brasil 4365
21040
Rio de Janeiro, RJ, Brazil.

 

 

Recebido para publicação em 16/01/91.

 

 

Departamento of Virology, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brazil.

Creative Commons License Todo o conteúdo deste periódico, exceto onde está identificado, está licenciado sob uma Licença Creative Commons