Measles serodiagnosis: standardization and evaluation of a dot-ELISA

Vaz de Lima, Lourdes R. A.; Hoshino-Shimizu, Sumie; Souza, Vanda A. U. F. de; Pannuti, Claudio S.; Andrade Junior, Heitor F.; Sumita, Laura M.; Ferreira, Antonio W.

doi:10.1590/S0036-46651994000200008

Abstracts

A Dot-ELISA using a measles virus (MV) antigen obtained by sodium deoxycholate treatment was standardized and evaluated for IgM and IgG antibody detection in measles patients and measles-vaccinated subjects. A total of 192 serum samples were studied, comprising 47 from patients with acute and convalescent measles, 55 from 9-month old children prior to measles vaccination and 41 from children of the same age after vaccination, and 49 from patients with unrelated diseases. The diagnostic performances of the IgG Dot-ELISA and IgG immuno fluorescence test (IFT) were found to be close, varying from 0.97 to 1.00 in sensitivity and the specificities were maximum (1.00). Nevertheless, the sensitivity of the IgM Dot-ELISA (0.85) was higher than that (0.63) of the IgM IFT, although both assays had comparably high (1.00) specificities. The IgM Dot-ELISA in particular proved to be more sensitive in relation to other assays studied by revealing antibodies in 80.0% (12/15) of vaccinated children on the 15th day after immunization. In contrast the IgM IFT, failed to detect antibodies in the same group of vaccinated children. The stability of the MV antigen was longer than that of the IFT antigen, and the reproducibility of the Dot-Elisa was satisfactory.

Dot-ELISA; Measles; Serologic diagnosis

A técnica de Dot-ELISA (DE) para detecção de anticorpos IgM e IgG anti vírus do sarampo foi padronizada e avaliada utilizando-se antígeno viral obtido por tratamento com desoxicolato de sódio (DOC). Foram estudadas 192 amostras de soros, compreendendo 47 amostras de 22 pacientes com sarampo nas fases aguda e convalescente, 55 amostras de soros de crianças antes da vacinação, tendo 9 meses de idade, 41 amostras de soros de crianças da mesma idade colhidas após vacinação e 49 amostras de soros de pacientes com outras patologias. O desempenho diagnóstico da técnica de Dot-ELISA-IgG foi semelhante ao de Imunofluorescência indireta (IFI) IgG cujos índices de sensibilidade variaram de 0,97 a 1,00 e os de especificidade sendo de valor máximo, 1,00. Contudo, a sensibilidade da técnica de Dot-ELISA IgM (0,85) foi mais alta que a de IFT IgM (0,63), embora ambos os ensaios apresentassem especificidades máximas (1,00). A técnica de Dot-ELISA IgM em particular mostrou-se mais sensível em relação a IFT, revelando anticorpos em 80% das crianças vacinadas (12/15), 15 dias após a imunização. Ao contrário, IFT IgM falhou em detectar anticorpos no mesmo grupo de crianças vacinadas. A estabilidade do antígeno viral obtido com detergente (DOC) foi maior que a do antígeno de IFT, e a reprodutibilidade da técnica de Dot-ELISA foi satisfatória.

SEROLOGY

Measles serodiagnosis: standardization and evaluation of a dot-ELISA

Diagnóstico sorológico do sarampo: padronização e avaliação do teste Dot-ELISA

Lourdes R. A. Vaz de Lima^I; Sumie Hoshino-Shimizu^I; Vanda A. U. F. de Souza^II; Claudio S. Pannuti^II; Heitor F. Andrade Junior^II; Laura M. Sumita^II; Antonio W. Ferreira^II

^ISeção de Imunologia, Divisão de Biologia Médica, Instituto Adolfo Lutz. 01246-902. São Paulo, SP, Brazil

^IILaboratório de Virologia e Soroepidemiologia do Instituto de Medicina Tropical de São Paulo, SP, Brasil

^{Address for correspondence} Address for correspondence: Lourdes R. A. Vaz de Lima Seção de Imunologia, Instituto Adolfo Lutz Av. Dr. Arnaldo 351 01246-902. São Paulo, SP, Brazil

SUMMARY

A Dot-ELISA using a measles virus (MV) antigen obtained by sodium deoxycholate treatment was standardized and evaluated for IgM and IgG antibody detection in measles patients and measles-vaccinated subjects. A total of 192 serum samples were studied, comprising 47 from patients with acute and convalescent measles, 55 from 9-month old children prior to measles vaccination and 41 from children of the same age after vaccination, and 49 from patients with unrelated diseases. The diagnostic performances of the IgG Dot-ELISA and IgG immuno fluorescence test (IFT) were found to be close, varying from 0.97 to 1.00 in sensitivity and the specificities were maximum (1.00). Nevertheless, the sensitivity of the IgM Dot-ELISA (0.85) was higher than that (0.63) of the IgM IFT, although both assays had comparably high (1.00) specificities. The IgM Dot-ELISA in particular proved to be more sensitive in relation to other assays studied by revealing antibodies in 80.0% (12/15) of vaccinated children on the 15th day after immunization. In contrast the IgM IFT, failed to detect antibodies in the same group of vaccinated children. The stability of the MV antigen was longer than that of the IFT antigen, and the reproducibility of the Dot-Elisa was satisfactory.

Keywords: Dot-ELISA; Measles, vaccination; Serologic diagnosis.

RESUMO

A técnica de Dot-ELISA (DE) para detecção de anticorpos IgM e IgG anti vírus do sarampo foi padronizada e avaliada utilizando-se antígeno viral obtido por tratamento com desoxicolato de sódio (DOC).

Foram estudadas 192 amostras de soros, compreendendo 47 amostras de 22 pacientes com sarampo nas fases aguda e convalescente, 55 amostras de soros de crianças antes da vacinação, tendo 9 meses de idade, 41 amostras de soros de crianças da mesma idade colhidas após vacinação e 49 amostras de soros de pacientes com outras patologias.

O desempenho diagnóstico da técnica de Dot-ELISA-IgG foi semelhante ao de Imunofluorescência indireta (IFI) IgG cujos índices de sensibilidade variaram de 0,97 a 1,00 e os de especificidade sendo de valor máximo, 1,00. Contudo, a sensibilidade da técnica de Dot-ELISA IgM (0,85) foi mais alta que a de IFT IgM (0,63), embora ambos os ensaios apresentassem especificidades máximas (1,00).

A técnica de Dot-ELISA IgM em particular mostrou-se mais sensível em relação a IFT, revelando anticorpos em 80% das crianças vacinadas (12/15), 15 dias após a imunização. Ao contrário, IFT IgM falhou em detectar anticorpos no mesmo grupo de crianças vacinadas. A estabilidade do antígeno viral obtido com detergente (DOC) foi maior que a do antígeno de IFT, e a reprodutibilidade da técnica de Dot-ELISA foi satisfatória.

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Recebido para publicação em 12/08/1993.

Aceito para publicação em 24/09/1993.

1. AEPPLI, R.E.; BARGETZI, M.J. & BINZ, H. - Diagnosis screening of multiple antigen-antibody reactions in a new single assay on nitrocellulose: the line immunobinding assay (LIBA). J. immunol. Meth., 120: 93-98, 1989.
2. BENNETT, F.C. & YEOMAN, L.C. - An improved procedure for the dot-immunobinding analysis of hybridoma supernatants. J. immunol. Meth., 61: 201-207, 1983.
3. BRADFORD, M.M. - A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Analyt. Biochem., 72: 248-254, 1976.
4. CENTERS FOR DISEASE CONTROL - MEASLES-United States, 1989, and first 20 weeks. MMWR, 30: 353-363, 1990.
5. CREMER, N.E.; COSSEN, C.K.; SHELL, G. et al. - 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 infection with those viruses. J. clin. Microbiol., 21: 869-874, 1985.
6. ERDMAN, D.D.; ANDERSON, L.J.; ADAMS, D.R. et al. - Evaluation of monoclonal antibody-based Capture Enzyme Immunoassays for detection of specific antibodies to measles virus. J. clin. Microbiol., 29: 1466-1471, 1991.
7. FLEISS, A.R. - Clinical epidemiology. In: FLEISS, A.R. The architecture of clinical research. Philadelphia, W. B. Saunders, 1985. p. 185-186.
8. GALEN, R.S. & GAMBINO, S.R. - Beyond normality: the predictive value and efficiency of medical diagnosis. New York, John Wiley, 1975.
9. GARDAS, A. & LEWAARTOWSKA, A. - Coating of protein to polystyrene ELISA plates in the presence of detergents. J. immunol. Meth., 106: 251-255, 1988.
10. HAWKES, R.; NIDAY, E. & GORDON, J. - A Dot Immunobinding Assay for monoclonal and other antibodies. Analyt. Biochem., 119: 142-147, 1982.
11. HJELMELAND, L.M. - A nondenaturing zwitterionic detergent for membrane biochemistry: design and synthesis. Proc. nat. Acad. Sci. (Wash.), 77: 6368-6370, 1980.
12. KALTER, S.S.; HEBERLING, R.L. & BARRY, J.D. - Detection and titration of measles virus antibody by hemagglutination inhibition and by Dot Immunobinding. J. clin. Microbiol., 29: 202-204, 1991.
13. KANAMURA, H.Y.; HOSHINO-SHIMIZU, S. & SILVA, L.C. - Schistosoma mansoni cercaría and schistosomulum antigens in serodiagnosis of schistosomiasis. Bull. Pan Amer. Hlth. Org., 26: 220-229, 1992.
14. LABETA, M.O.; FERNANDEZ, N. & FESTENSTEIN, H. - Solubilization effect of Nonidet P40, Triton X-100 and CHAPS in the detection of MCH-like glycoproteins. J. immunol. Meth., 112: 133-138, 1988.
15. LOWRY, O.H.; ROSEBROUGH, H.J.; FARR, L. & RANDALL, R.J. - Protein measurement with the Folin phenol reagent. J. biol. Chem., 193: 265-275, 1951.
16. MACLURE, M. & WILLET, W.C. - Misinterpretation and misuse of Kappa statistic. Amer. J. Epidem., 126: 161-169, 1987.
17. MARKOWITZ, L.E.; PREBLUD, S.R.; FINE, P.E.M. & ORENSTEIN, W.A. - Duration of live measles vaccine-induced immunity. Pediat. infect. Dis., 9: 101-110, 1987.
18. MAYO, D.R. BRENNAN., T.; CORMIER, D.P.; HADLER, J. & LAMB, P. - Evaluation of a Commercial Measles virus Immunoglobulin M Enzyme Immunoassay. J. clin. Microbiol., 29: 2865-2867, 1990.
19. ROSSIER, E.; MILLER, H.; McCULLOCH, B.; SULLIVAN, L. & WARD, K. - Comparison of Immunofluorescence and Enzyme Immunoassay for detection of measles specific Immunoglobulin M antibody. J. clin. Microbiol., 29: 1069-1071, 1991.
20. SOUZA, V.A.U.F.; PANNUTI, C.S.; SUMITA, L.M. & ALBRECHT, P. - Enzyme-Linked Immunosorbent Assay (ELISA) for measles antibody. A comparison with haemagglutination inhibition, immunofluorescence and plaque neutralizations test. Rev. Inst. Med. trop. S. Paulo, 33: 32-36, 1991.

Address for correspondence:

Lourdes R. A. Vaz de Lima

Seção de Imunologia, Instituto Adolfo Lutz

Av. Dr. Arnaldo 351

01246-902. São Paulo, SP, Brazil

Publication Dates

Publication in this collection
20 Sept 2006
Date of issue
Apr 1994

History

Accepted
24 Sept 1993
Received
12 Aug 1993

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

[1] 1. AEPPLI, R.E.; BARGETZI, M.J. & BINZ, H. - Diagnosis screening of multiple antigen-antibody reactions in a new single assay on nitrocellulose: the line immunobinding assay (LIBA). J. immunol. Meth., 120: 93-98, 1989.

[2] 2. BENNETT, F.C. & YEOMAN, L.C. - An improved procedure for the dot-immunobinding analysis of hybridoma supernatants. J. immunol. Meth., 61: 201-207, 1983.

[3] 3. BRADFORD, M.M. - A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Analyt. Biochem., 72: 248-254, 1976.

[4] 4. CENTERS FOR DISEASE CONTROL - MEASLES-United States, 1989, and first 20 weeks. MMWR, 30: 353-363, 1990.

[5] 5. CREMER, N.E.; COSSEN, C.K.; SHELL, G. et al. - 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 infection with those viruses. J. clin. Microbiol., 21: 869-874, 1985.

[6] 6. ERDMAN, D.D.; ANDERSON, L.J.; ADAMS, D.R. et al. - Evaluation of monoclonal antibody-based Capture Enzyme Immunoassays for detection of specific antibodies to measles virus. J. clin. Microbiol., 29: 1466-1471, 1991.

[7] 7. FLEISS, A.R. - Clinical epidemiology. In: FLEISS, A.R. The architecture of clinical research. Philadelphia, W. B. Saunders, 1985. p. 185-186.

[8] 8. GALEN, R.S. & GAMBINO, S.R. - Beyond normality: the predictive value and efficiency of medical diagnosis. New York, John Wiley, 1975.

[9] 9. GARDAS, A. & LEWAARTOWSKA, A. - Coating of protein to polystyrene ELISA plates in the presence of detergents. J. immunol. Meth., 106: 251-255, 1988.

[10] 10. HAWKES, R.; NIDAY, E. & GORDON, J. - A Dot Immunobinding Assay for monoclonal and other antibodies. Analyt. Biochem., 119: 142-147, 1982.

[11] 11. HJELMELAND, L.M. - A nondenaturing zwitterionic detergent for membrane biochemistry: design and synthesis. Proc. nat. Acad. Sci. (Wash.), 77: 6368-6370, 1980.

[12] 12. KALTER, S.S.; HEBERLING, R.L. & BARRY, J.D. - Detection and titration of measles virus antibody by hemagglutination inhibition and by Dot Immunobinding. J. clin. Microbiol., 29: 202-204, 1991.

[13] 13. KANAMURA, H.Y.; HOSHINO-SHIMIZU, S. & SILVA, L.C. - Schistosoma mansoni cercaría and schistosomulum antigens in serodiagnosis of schistosomiasis. Bull. Pan Amer. Hlth. Org., 26: 220-229, 1992.

[14] 14. LABETA, M.O.; FERNANDEZ, N. & FESTENSTEIN, H. - Solubilization effect of Nonidet P40, Triton X-100 and CHAPS in the detection of MCH-like glycoproteins. J. immunol. Meth., 112: 133-138, 1988.

[15] 15. LOWRY, O.H.; ROSEBROUGH, H.J.; FARR, L. & RANDALL, R.J. - Protein measurement with the Folin phenol reagent. J. biol. Chem., 193: 265-275, 1951.

[16] 16. MACLURE, M. & WILLET, W.C. - Misinterpretation and misuse of Kappa statistic. Amer. J. Epidem., 126: 161-169, 1987.

[17] 17. MARKOWITZ, L.E.; PREBLUD, S.R.; FINE, P.E.M. & ORENSTEIN, W.A. - Duration of live measles vaccine-induced immunity. Pediat. infect. Dis., 9: 101-110, 1987.

[18] 18. MAYO, D.R. BRENNAN., T.; CORMIER, D.P.; HADLER, J. & LAMB, P. - Evaluation of a Commercial Measles virus Immunoglobulin M Enzyme Immunoassay. J. clin. Microbiol., 29: 2865-2867, 1990.

[19] 19. ROSSIER, E.; MILLER, H.; McCULLOCH, B.; SULLIVAN, L. & WARD, K. - Comparison of Immunofluorescence and Enzyme Immunoassay for detection of measles specific Immunoglobulin M antibody. J. clin. Microbiol., 29: 1069-1071, 1991.

[20] 20. SOUZA, V.A.U.F.; PANNUTI, C.S.; SUMITA, L.M. & ALBRECHT, P. - Enzyme-Linked Immunosorbent Assay (ELISA) for measles antibody. A comparison with haemagglutination inhibition, immunofluorescence and plaque neutralizations test. Rev. Inst. Med. trop. S. Paulo, 33: 32-36, 1991.