Comparison of indirect immunofluorescence test for measles antibodies with haemagglutination inhibition and plaque neutralization tests

Souza, Vanda Akico Ueda Fick de; Pannuti, Cláudio Sérgio; Godoy, Cid Vieira Franco de; Albrecht, Paul; Lopes, Marta Heloísa; Mendonça, João da Silva

doi:10.1590/S0036-46651990000500009

Abstracts

Indirect Immunofluorescence (IFA), Plaque Reduction Neutralization (PRN) and Haemagglutination Inhibition (HI) tests for measles antibodies were carried out in 197 sera obtained from umbilical cord and vaccinated children. The IFA was also applied to blood samples collected with filter paper. IFA results demonstrated that the test is relatively simple to perform, with good reproducibility for different antigen lots. Good correlation was obtained between IFA, PRN and HI antibody titers. Better correlation was demonstrated with IFA and PRN than with HI and PRN tests. Sensitivity of IFA in detecting antibody was less effective than PRN, however more effective than HI using rhesus monkey red blood cells. PRN antibody titers over 100 were detected by IFA but not by HI (9.7% with negative results). IFA may be of considerable practical use and able to substitute HI in Seroepidemiological surveys and to evaluate vaccine efficacy. It also can be simplified by employing filter paper collected samples.

Measles; Indirect immunofluorescence; Plaque reduction; Haemagglutination inhibiton

As reações de imunofluorescência indireta (RIF), neutralização por redução de placas (RNP) e inibição da hemaglutinação (RIH) para detecção de anticorpos para o vírus do sarampo foram aplicadas a 197 soros provenientes de cordão umbilical e de crianças vacinadas contra o sarampo. Avaliou-se ainda a aplicação da RIF em amostras colhidas em papel de filtro. A RIF apresentou-se como uma prova de execução relativamente simples, de boa reprodutibilidade com diferentes partidas de antígeno. Observou-se boa correlação entre os títulos de anticorpos obtidos por RIF, RNP e RIH. Com a RNP, a RIF apresentou maior correlação que a RIH. A sensibilidade da RIF na detecção de anticorpos contra o sarampo foi menor que RNP, porém superior à da RIH com hemácias de macaco rhesus. Anticorpos com títulos superiores a 100 pela RNP foram sistematicamente detectados pela RIF, mas não por RIH (9,7% de resultados negativos). A RIF pode ser de grande utilidade, podendo substituir com vantagem a RIH em inquéritos soroepidemiológicos e na avaliação de eficácia de vacinas. Além disso, mostrou-se adequada para aplicação em larga escala, já que permite processar amostras de sangue colhidas em papel de filtro.

SEROLOGY

Comparison of indirect immunofluorescence test for measles antibodies with haemagglutination inhibition and plaque neutralization tests

Comparação da reação de imunofluorescência indireta para o vírus do sarampo com as reações de inibição da hemaglutinação e neutralização por redução de placas

Vanda Akico Ueda Fick de Souza^I; Cláudio Sérgio Pannuti^I; Cid Vieira Franco de Godoy^II; Paul Albrecht^III; Marta Heloísa Lopes^II; João da Silva Mendonça^IV

^IInstituto de Medicina Tropical de São Paulo (LIM/52-HC-FM-USP)

^IIFaculdade de Medicina da Universidade de São Paulo

^IIIDivision of Virology, FDA, Bethesda

^IVHospital do Servidor Público do Estado de São Paulo

^{Address for correspondence} Address for correspondence: Vanda Akico Ueda Fick de Souza Instituto de Medicina Tropical de São Paulo Av. Dr. Enéas de Carvalho Aguiar, 470 05403 São Paulo, SP, Brasil

SUMMARY

Indirect Immunofluorescence (IFA), Plaque Reduction Neutralization (PRN) and Haemagglutination Inhibition (HI) tests for measles antibodies were carried out in 197 sera obtained from umbilical cord and vaccinated children. The IFA was also applied to blood samples collected with filter paper.

IFA results demonstrated that the test is relatively simple to perform, with good reproducibility for different antigen lots. Good correlation was obtained between IFA, PRN and HI antibody titers. Better correlation was demonstrated with IFA and PRN than with HI and PRN tests.

Sensitivity of IFA in detecting antibody was less effective than PRN, however more effective than HI using rhesus monkey red blood cells. PRN antibody titers over 100 were detected by IFA but not by HI (9.7% with negative results). IFA may be of considerable practical use and able to substitute HI in Seroepidemiological surveys and to evaluate vaccine efficacy. It also can be simplified by employing filter paper collected samples.

Key words: Measles; Indirect immunofluorescence; Plaque reduction, Haemagglutination inhibiton.

RESUMO

As reações de imunofluorescência indireta (RIF), neutralização por redução de placas (RNP) e inibição da hemaglutinação (RIH) para detecção de anticorpos para o vírus do sarampo foram aplicadas a 197 soros provenientes de cordão umbilical e de crianças vacinadas contra o sarampo. Avaliou-se ainda a aplicação da RIF em amostras colhidas em papel de filtro.

A RIF apresentou-se como uma prova de execução relativamente simples, de boa reprodutibilidade com diferentes partidas de antígeno. Observou-se boa correlação entre os títulos de anticorpos obtidos por RIF, RNP e RIH. Com a RNP, a RIF apresentou maior correlação que a RIH.

A sensibilidade da RIF na detecção de anticorpos contra o sarampo foi menor que RNP, porém superior à da RIH com hemácias de macaco rhesus.

Anticorpos com títulos superiores a 100 pela RNP foram sistematicamente detectados pela RIF, mas não por RIH (9,7% de resultados negativos). A RIF pode ser de grande utilidade, podendo substituir com vantagem a RIH em inquéritos soroepidemiológicos e na avaliação de eficácia de vacinas. Além disso, mostrou-se adequada para aplicação em larga escala, já que permite processar amostras de sangue colhidas em papel de filtro.

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Recebido para publicação em 23/11/1989.

1. ALBRECHT, P.; ENNIS, F.A.; SALTZMAN, E.J. & KRUGMAN, S. Persistence of maternal antibody in infants beyond 12 months: mechanism of measles vaccine failure. J. Pediat., 91: 715-718, 1977.
2. ALBRECHT, P.; HERRMANN, K. & BURNS, G.R. Role of virus strain in conventional and enhanced measles plaque neutralization test. J. virol. Meth., 3: 251-260, 1981.
3. BIANO, S.A.; CHANG, T.W. & DANIELS, J.B. Rubella haemagglutination inhibition removal of nonspecific agglutination due to manganous chloride. Appl. Microbiol., 28: 992-994, 1974.
4. CONNOLLY, J.H.; HAIRE, M. & HADDEN, D.S.M. - Measles immunoglobulins in subacute sclerosing panencephalitis. Brit. med. J., 1: 23-25, 1971.
5. CREMER, N.E.; COSSEN, C.K.; SHELL, G.; DIGGS, J.; GALLO, D. & SCHMIDT, N.J. Enzyme immunoassay versus plaque neutralization and other methods for determination of immune status to measles and varicella-zoster viruses and versus complement fixation for serodiagnosis of infections with those viruses. J. clin. Microbiol., 21: 869-874, 1985.
6. CUTCHINS, E.C. A comparison of the haemagglutination-inhibition, neutralization and complement fixation tests in the assay of antibody to measles. J. Immunol., 88: 788-795, 1962.
7. EMMONS, R.W. & RIGGS, J.L. Application of immunofluorescence to diagnosis of viral infections. In: MARAMOROSCH, K. & KOPROWSKY, H. Methods in virology. New York, Academic Press, 1977. v. 6. p. 1-28.
8. GERSHON, A.A. & KRUGMAN, S. Measles virus. In: LENNETTE, E.H. & SCHMIDT, N.S. Diagnostics procedures for viral and rickettsial infections. 5. ed. New York, Am. Public. Health Assoc., 1979. p. 665-693.
9. KLEIMAN, M.B.; BLACKBURN, C.K.L.; ZIMMERMAN, S.E. & FRENCH, M.L.V. Comparison of enzyme-linked immunosorbent assay for acute measles with hemagglutination inhibition, complement fixation, and fluorescent-antibody methods. J. clin. Microbiol., 14: 147-152, 1981.
10. NAGY, G.; KÓSA, S.; TAKÁTSY, S. & KOLLER, M. The use of IgM tests for analysis of the causes of measles vaccine failures: experience gained in an epidemic in Hungary in 1980 and 1981. J. med. Virol., 13: 93-103, 1984.
11. NAKANO, J.H.; MILLER, D.L.; FOSTER, S.O. & BRINK, E.W. Microtiter determination of measles haemaggiutination inhibition antibody with filter paper. J. clin. Microbiol., 17: 860-863, 1983.
12. SOUZA, S.L. & CAMARGO, M.E. The use of filter paper blood smears in a practical fluorescent test for American trypanosomiasis serodiagnosis. Rev. Inst. Med. trop. S. Paulo, 8: 255-258, 1966.
13. WASSILAK, S.G.F.; BERNIER, R.H.; HERRMANN, K.L.; ORESTEIN, W.A.; BART, K.J. & AMLER, R. Measles seroconfirmation using dried capillary blood specimens in filter paper. Pediat. infect. Dis., 3: 117-121, 1984.
14. YEAGER, A.S.; DAVIS, J.H.; ROSS, L.A. & HARVEY, B. Measles immunization. Successes and failures. J. Amer. med. Ass., 237: 347-351, 1977.

Address for correspondence:

Vanda Akico Ueda Fick de Souza

Instituto de Medicina Tropical de São Paulo

Av. Dr. Enéas de Carvalho Aguiar, 470

05403 São Paulo, SP, Brasil

Publication Dates

Publication in this collection
12 Sept 2006
Date of issue
Oct 1990

History

Received
23 Nov 1989

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

[1] 1. ALBRECHT, P.; ENNIS, F.A.; SALTZMAN, E.J. & KRUGMAN, S. Persistence of maternal antibody in infants beyond 12 months: mechanism of measles vaccine failure. J. Pediat., 91: 715-718, 1977.

[2] 2. ALBRECHT, P.; HERRMANN, K. & BURNS, G.R. Role of virus strain in conventional and enhanced measles plaque neutralization test. J. virol. Meth., 3: 251-260, 1981.

[3] 3. BIANO, S.A.; CHANG, T.W. & DANIELS, J.B. Rubella haemagglutination inhibition removal of nonspecific agglutination due to manganous chloride. Appl. Microbiol., 28: 992-994, 1974.

[4] 4. CONNOLLY, J.H.; HAIRE, M. & HADDEN, D.S.M. - Measles immunoglobulins in subacute sclerosing panencephalitis. Brit. med. J., 1: 23-25, 1971.

[5] 5. CREMER, N.E.; COSSEN, C.K.; SHELL, G.; DIGGS, J.; GALLO, D. & SCHMIDT, N.J. Enzyme immunoassay versus plaque neutralization and other methods for determination of immune status to measles and varicella-zoster viruses and versus complement fixation for serodiagnosis of infections with those viruses. J. clin. Microbiol., 21: 869-874, 1985.

[6] 6. CUTCHINS, E.C. A comparison of the haemagglutination-inhibition, neutralization and complement fixation tests in the assay of antibody to measles. J. Immunol., 88: 788-795, 1962.

[7] 7. EMMONS, R.W. & RIGGS, J.L. Application of immunofluorescence to diagnosis of viral infections. In: MARAMOROSCH, K. & KOPROWSKY, H. Methods in virology. New York, Academic Press, 1977. v. 6. p. 1-28.

[8] 8. GERSHON, A.A. & KRUGMAN, S. Measles virus. In: LENNETTE, E.H. & SCHMIDT, N.S. Diagnostics procedures for viral and rickettsial infections. 5. ed. New York, Am. Public. Health Assoc., 1979. p. 665-693.

[9] 9. KLEIMAN, M.B.; BLACKBURN, C.K.L.; ZIMMERMAN, S.E. & FRENCH, M.L.V. Comparison of enzyme-linked immunosorbent assay for acute measles with hemagglutination inhibition, complement fixation, and fluorescent-antibody methods. J. clin. Microbiol., 14: 147-152, 1981.

[10] 10. NAGY, G.; KÓSA, S.; TAKÁTSY, S. & KOLLER, M. The use of IgM tests for analysis of the causes of measles vaccine failures: experience gained in an epidemic in Hungary in 1980 and 1981. J. med. Virol., 13: 93-103, 1984.

[11] 11. NAKANO, J.H.; MILLER, D.L.; FOSTER, S.O. & BRINK, E.W. Microtiter determination of measles haemaggiutination inhibition antibody with filter paper. J. clin. Microbiol., 17: 860-863, 1983.

[12] 12. SOUZA, S.L. & CAMARGO, M.E. The use of filter paper blood smears in a practical fluorescent test for American trypanosomiasis serodiagnosis. Rev. Inst. Med. trop. S. Paulo, 8: 255-258, 1966.

[13] 13. WASSILAK, S.G.F.; BERNIER, R.H.; HERRMANN, K.L.; ORESTEIN, W.A.; BART, K.J. & AMLER, R. Measles seroconfirmation using dried capillary blood specimens in filter paper. Pediat. infect. Dis., 3: 117-121, 1984.

[14] 14. YEAGER, A.S.; DAVIS, J.H.; ROSS, L.A. & HARVEY, B. Measles immunization. Successes and failures. J. Amer. med. Ass., 237: 347-351, 1977.