Accessibility / Report Error

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

Abstracts

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.

Enzyme immunoassay; Anti-HAV; Hepatitis A


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.

Enzyme immmunoassay; Anti-HAV; Hepatitis A


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

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.

Recebido para publicação em 16/01/91.

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

  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 16. Parry JV. Hepatitis A infection: guidelines for development of satisfactory assays for laboratory diagnosis. Medical Laboratory Sciences 38: 303-311, 1981.
  • 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.
  • 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.
  • 19. Purcell RH, Wong DC, Alter HJ, Holland PV. Microtiter solid-phase radioimmunoassay for hepatitis B antigen. Applied Microbiology 26: 478-484, 1973.
  • 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.
  • 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.
  • 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.
  • 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.
  • Adress to correspondence:

    Dra. Claudia Lamarca Vitral.
    Depto. of Virology/FIOCRUZ.
    Av. Brasil 4365
    21040
    Rio de Janeiro, RJ, Brazil.
  • Publication Dates

    • Publication in this collection
      15 May 2013
    • Date of issue
      June 1991

    History

    • Received
      16 Jan 1991
    • Accepted
      16 Jan 1991
    Sociedade Brasileira de Medicina Tropical - SBMT Sociedade Brasileira de Medicina Tropical - SBMT, Núcleo de Medicina Tropical – UnB, Sala 43C – 70904-970, E-mails: rsbmt@uftm.edu.br | artes.rsbmt@gmail.com | sbmt@sbmt.org.br , WhatsApp: SBMT (61) 9.9192-6496, WhatsApp: RSBMT (34) 9.9996-5807 - Brasília - DF - Brazil
    E-mail: rsbmt@uftm.edu.br