Evaluation of the enzyme-linked-immuno-electro-diffusion-assay (ELIEDA) for the diagnosis of Schistosoma mansoni infection with low worm burden

Leal-Bacelar, Grace Mary; Kanamura, Hermínia Y.; Hoshino-Shimizu, Sumie; Lima, Dirce Mary C.; Abrantes-Lemos, Clarice P.; Silva, Luiz Caetano da

doi:10.1590/S0036-46651995000200006

Abstracts

An immunoprecipitation technique, ELIEDA (enzyme-linked-immuno-electro-diffusion assay), was evaluated for the diagnosis of Schistosoma mansoni infection with low worm burden. One hundred of serum samples from patients excreting less than 600 eggs per gram of feces (epg), with unrelated diseases and clinically healthy subjects were studied. In patients with egg counts higher than 200 epg, the sensitivities of IgM and IgG ELIEDA were 1.000 and 0.923, respectively, not differing from other Serologic techniques, such as indirect hemaglutination (IHAT), immunofluorescence (IFT) tests and immuno-electrodiffusion assay (IEDA). However in patients with low egg counts (< 100 epg), the IgG ELIEDA provided better results (0.821) than IgM ELIEDA (0.679), showing sensitivity that did not differ from that of IgG IFT (0.929), but lower than that of IgM IFT (0.964). However, its sensivity was higher than that found with IHAT (0.607) and IEDA (0.536). The specificity of IgG ELIEDA was comparable to that of other techniques. The data indicate that IgG ELIEDA might be useful for the diagnosis of slight S. mansoni infections, and the cellulose acetate membrane strips can be stored for further retrospective studies.

Schistosoma mansoni; Immunodiagnosis; ELIEDA

A técnica de imunoprecipitação ELIEDA (enzyme-linked-immuno-electro-diffusion-assay) foi avaliada para fins diagnósticos da esquistossomose mansoni em pacientes com baixa carga parasitária. Amostras de soros de 50 pacientes com exame de fezes positivo para S. mansoni (carga parasitária < 600 ovos por grama de fezes = opg) e 50 não esquistossomóticos (30 com outras afecções e 20 normais) foram estudadas. Em pacientes com carga parasitária acima de 200 opg, a sensibilidade da técnica de ELIEDA, tanto para anticorpos IgG como IgM, respectivamente 1,000 e 0,923, não diferiu da observada para outras reações sorológicas, como a de hemaglutinação (RHA), imunofluorescênca (RIF) e imunoeletrodifusão (IED). Entretanto, naqueles com baixa carga (< 100 opg), a ELIEDA-IgG mostrou resultados mais satisfatórios (0,821) que a ELIEDA-IgM (0,679), apresentando sensibilidade que não diferiu à da RIF-IgG (0,929); apesar de inferior à da RIF-IgM (0,964), a sensibilidade da ELIEDA-IgG foi superior à da RHA (0,607) e à da IED (0,536). Quanto à especificidade, esta foi comparável à dos demais testes estudados. Os dados indicam que a ELIEDA-IgG pode ser útil para diagnóstico da esquistossomose, mesmo em pacientes com pequena carga parasitária, com a vantagem de permitir estudos retrospectivos através da análise posterior das fitas de acetato de celulose que podem ser armazenadas e arquivadas.

DIAGNOSTIC STUDIES

Evaluation of the enzyme-linked-immuno-electro-diffusion-assay (ELIEDA) for the diagnosis of Schistosoma mansoni infection with low worm burden

Avaliação da técnica de ELIEDA (enzyme-linked-immuno-electro-diffusion-assay) para o diagnóstico da infecção pelo Schistosoma mansoni de baixa carga

Grace Mary Leal-Bacelar^I; Hermínia Y. Kanamura^I; Sumie Hoshino-Shimizu^II; Dirce Mary C. Lima^III; Clarice P. Abrantes-Lemos^III; Luiz Caetano da Silva^III

^IDepartment of Clinical and Toxicological Analyses, Faculty of Pharmaceutical Science, University of São Paulo, São Paulo, Brazil

^IIInstitute Adolfo Lutz, São Paulo State Health Department, São Paulo, Brazil

^IIIInstitute of Tropical Medicine of São Paulo, São Paulo, Brazil, (LIM/HCFMUSP-06)

^{Correspondence to} Correspondence to: Sumie Hoshino-Shimizu Instituto Adolfo Lutz - Immunology Section Av. Dr. Arnaldo, 355 -11. andar 01246-902 São Paulo, SP, Brazil

SUMMARY

An immunoprecipitation technique, ELIEDA (enzyme-linked-immuno-electro-diffusion assay), was evaluated for the diagnosis of Schistosoma mansoni infection with low worm burden. One hundred of serum samples from patients excreting less than 600 eggs per gram of feces (epg), with unrelated diseases and clinically healthy subjects were studied. In patients with egg counts higher than 200 epg, the sensitivities of IgM and IgG ELIEDA were 1.000 and 0.923, respectively, not differing from other Serologic techniques, such as indirect hemaglutination (IHAT), immunofluorescence (IFT) tests and immuno-electrodiffusion assay (IEDA). However in patients with low egg counts (< 100 epg), the IgG ELIEDA provided better results (0.821) than IgM ELIEDA (0.679), showing sensitivity that did not differ from that of IgG IFT (0.929), but lower than that of IgM IFT (0.964). However, its sensivity was higher than that found with IHAT (0.607) and IEDA (0.536). The specificity of IgG ELIEDA was comparable to that of other techniques. The data indicate that IgG ELIEDA might be useful for the diagnosis of slight S. mansoni infections, and the cellulose acetate membrane strips can be stored for further retrospective studies.

Keywords:Schistosoma mansoni; Immunodiagnosis; ELIEDA.

RESUMO

A técnica de imunoprecipitação ELIEDA (enzyme-linked-immuno-electro-diffusion-assay) foi avaliada para fins diagnósticos da esquistossomose mansoni em pacientes com baixa carga parasitária. Amostras de soros de 50 pacientes com exame de fezes positivo para S. mansoni (carga parasitária < 600 ovos por grama de fezes = opg) e 50 não esquistossomóticos (30 com outras afecções e 20 normais) foram estudadas. Em pacientes com carga parasitária acima de 200 opg, a sensibilidade da técnica de ELIEDA, tanto para anticorpos IgG como IgM, respectivamente 1,000 e 0,923, não diferiu da observada para outras reações sorológicas, como a de hemaglutinação (RHA), imunofluorescênca (RIF) e imunoeletrodifusão (IED). Entretanto, naqueles com baixa carga (< 100 opg), a ELIEDA-IgG mostrou resultados mais satisfatórios (0,821) que a ELIEDA-IgM (0,679), apresentando sensibilidade que não diferiu à da RIF-IgG (0,929); apesar de inferior à da RIF-IgM (0,964), a sensibilidade da ELIEDA-IgG foi superior à da RHA (0,607) e à da IED (0,536). Quanto à especificidade, esta foi comparável à dos demais testes estudados. Os dados indicam que a ELIEDA-IgG pode ser útil para diagnóstico da esquistossomose, mesmo em pacientes com pequena carga parasitária, com a vantagem de permitir estudos retrospectivos através da análise posterior das fitas de acetato de celulose que podem ser armazenadas e arquivadas.

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Recebido para publicação em 18/05/1994.

Aceito para publicação em 24/11/1994.

1. CHIEFFI, P.P. & KANAMURA, H.Y. - Diagnóstico laboratorial da esquistossomose mansônica. Rev. bras. Malar., 30: 77-97, 1978.
2. COLTORTI, E.A.; FERNANDEZ, E.; MARGUET, E.R. et al. - Detección de portadores asintomáticos de quistes hidáticos: aumento de la especificidad del ensayo inmuno enzimático. Rev. Inst. Med. trop. S. Paulo, 32: 275-284, 1990.
3. DEELDER, A.M.; KORNELIS, D.; RIER, W.B. et al. - Immunodiagnosis of human schistosomiasis using different immunoprecipitation techniques. Z. Parasitenk., 47: 111-118, 1975.
4. DEELDER, A.M.; KORNELIS, D.; MAKBIN, M. et al. - Applicability of different antigen preparations in the enzyme-linked immunosorbent assay for schistosomiasis mansoni. Amer. J. trop. Med. Hyg., 29: 401-410, 1980.
5. DIAS, L.C.S.; KANAMURA, H.Y.; HOSHINO-SHIMIZU, S. et al. - Field trials for immunodiagnosis with reference to Schistosoma mansoni. In: BERGQUIST, N.R. - Immunodiagnostic approaches in Schistosomiasis. New York, John Wiley & Sons, 1992. p. 39-47.
6. DUNNE, D.W.; JONES, F.M. & DOENHOFF, M.J. - The purification, characterization, serological activity and hepatotoxic properties of two cationic glycoproteins (alfa-1 and omega-1), from S. mansoni eggs. Parasitology, 103: 225-236, 1991.
7. EVENGARD, B.; HAMMARSTROM, L.; SMITH, C.J.E. et al. - Subclasss distribution and IgE responses after treatment in human schistosomiasis. Clin. exp. Immunol., 73: 383-388, 1988.
8. GALEN, R.S. & GAMBINO, S.R. - Beyond normality: the predictive value and efficiency of medical diagnosis. New York, John Wiley & Sons, 1975.
9. GENTILINI, M.; PINON, J.M. & NIEL, G. - Immunoélectrodiffusion sur membrane d'ácetate de cellulose. Applications en parasitologie ŕ propos de resultats préliminaires sur 600 essais. Nouv. Presse méd., 1: 1499-1501, 1972.
10. JASSIM, A.; HASSAN, K. & CATTY, D. - Antibody isotypes in human schistosomiasis mansoni. Paras. Immunol., 9: 627-650, 1987.
11. KANAMURA-KAWADA, H.Y. - Antígenos de diferentes formas evolutivas do Schistosoma mansoni para diagnóstico e acompanhamento sorológicos de pacientes submetidos ŕ quimioterapia. Săo Paulo, 1985. (Tese de Doutoramento - Instituto de Cięncias Biomédicas da Universidade de Săo Paulo).
12. KANAMURA, H.Y.; HOSHINO-SHIMIZU, S.; CAMARGO, M.E. et al. - Class specific antibodies and fluorescent staining patterns in acute and chronic forms of schistosomiasis mansoni. Amer. J. trop. Med. Hyg., 28: 242-248, 1979.
13. KANAMURA, H.Y.; HOSHINO-SHIMIZU, S. & SILVA, L.C. da - Solubilization of antigens of Schistosoma mansoni adult worms for the passive hemagglutination test. Preliminary reports. Rev. Inst. Med. trop. S. Paulo, 23: 92-95, 1981.
14. KANAMURA, H.Y.; HOSHINO-SHIMIZU, S. & SILVA, L.C. da - Schistosoma mansoni cercaría and schistosomulum antigens in serodiagnosis of Schistosomiasis. Bull. Pan. Amer. Hlth. Org. 26: 220-229, 1992.
15. KATZ, N.; CHAVES, A. & PELLEGRINO, J. - A simple device for quantitative stool thick smear technique in schistosomiasis mansoni. Rev. Inst. Med. trop. S. Paulo, 14: 397-400, 1972.
16. LOWRY, O.H.; ROSEBROUGH, N.J.; FARR, A.L. et al. - Protein measurement with the Folin phenol reagent. J. biol. Chem., 193: 265-271, 1951.
17. MADDISON, S.E.; SLEMENDA, S.B.; TSANG, V.C.W. et al. - Serodiagnosis of schistosomiasis mansoni with microsomal adult worm antigen in an enzyme-linked-immunosorbent-assay using a standard curve developed with a reference serum pool. Amer. J. trop. Med. Hyg., 34: 484-494, 1985.
18. MOTT, K.E. & DIXON, H. - Collaborative study on antigens for immunodiagnosis of schistosomiasis. Bull. Wld. Hlth. Org., 60: 729-753, 1982.
19. PINON, J.M. & DROPSY, G. - Étude immunologique des parasitoses humaines. Interęt de I'ELIEDA (Enzyme-linked-immuno-electro-diffusion-assay). Path. Biol., 25: 23-27, 1977.
20. PINON, J.M. & DROPSY, G. - ELIEDA (Enzyme-linked-immuno-electro-diffusion-assay) application of a combined immunoelectrodiffusion and immunoenzyme method to the study of immune response in parasitic infections. J. immunol. Meth., 16: 15-22, 1977.
21. PINON, J.M. & DROPSY, G. - Étude de l'ímmunité humorale dans la bilharziose humaine. Interęt de I'ELIEDA (Enzyme-linked-immuno-electro-diffusion-assay). Bull. Soc. Path. exot., 69: 238-244, 1977.
22. PINON, J.M.; GORSE, J.P. & DROPSY, G. - Exploration de l'ímmunité humorale dans L'aspergillose: intéręt de I'ELIEDA (enzyme-linked-immuno-electro-diffusion-assay). Mycopathologia (Den Haag), 60: 115-120, 1977.
23. PINON, J.M.; SULAHIAN, A. & DROPSY, G. - The use of ELIEDA (enzyme-linked-immuno-electro-diffusion-assay) in the study of humoral antibodies in human schistosomiasis. Trans. roy. Soc. trop. Med. Hyg., 72: 429-495, 1978.
24. PINON, J.M.; SULAHIAN, A.; REMY, G. et al. - Immunological study of hydatidosis. I. Evaluation of immunoelectrodiffusion tests and enzyme-linked-immuno-electro-diffusion-assay (ELIEDA) in human hydatidosis. Amer. J. trop. Med. Hyg., 28: 318-324, 1979.
25. PINON, J.M.; THOANNES, H. & GRUSON, N. - An enzyme-linked-immuno-filtration-assay used to compare infant and maternal antibody profiles in toxoplasmosis. J. immunol. Meth., 77: 15-23, 1985.
26. ROTHMAN, K.J. & BOICE, J.D. - Epidemiologic analysis with a programmable calculator. Boston, Epidemiology Resources Inc., 1982.
27. RUIZ-TIBEN, E.; HILLYER, G.V.; KNIGHT, W.B. et al. - Intensity of infection with S. mansoni: its relationship to the sensitivity and specificity of Serologic tests. Amer. J. trop. Med. Hyg., 28: 230-236, 1979.

Correspondence to:

Sumie Hoshino-Shimizu

Instituto Adolfo Lutz - Immunology Section

Av. Dr. Arnaldo, 355 -11. andar

01246-902 São Paulo, SP, Brazil

Publication Dates

Publication in this collection
22 Feb 2007
Date of issue
Apr 1995

History

Accepted
24 Nov 1994
Received
18 May 1994

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

[1] 1. CHIEFFI, P.P. & KANAMURA, H.Y. - Diagnóstico laboratorial da esquistossomose mansônica. Rev. bras. Malar., 30: 77-97, 1978.

[2] 2. COLTORTI, E.A.; FERNANDEZ, E.; MARGUET, E.R. et al. - Detección de portadores asintomáticos de quistes hidáticos: aumento de la especificidad del ensayo inmuno enzimático. Rev. Inst. Med. trop. S. Paulo, 32: 275-284, 1990.

[3] 3. DEELDER, A.M.; KORNELIS, D.; RIER, W.B. et al. - Immunodiagnosis of human schistosomiasis using different immunoprecipitation techniques. Z. Parasitenk., 47: 111-118, 1975.

[4] 4. DEELDER, A.M.; KORNELIS, D.; MAKBIN, M. et al. - Applicability of different antigen preparations in the enzyme-linked immunosorbent assay for schistosomiasis mansoni. Amer. J. trop. Med. Hyg., 29: 401-410, 1980.

[5] 5. DIAS, L.C.S.; KANAMURA, H.Y.; HOSHINO-SHIMIZU, S. et al. - Field trials for immunodiagnosis with reference to Schistosoma mansoni. In: BERGQUIST, N.R. - Immunodiagnostic approaches in Schistosomiasis. New York, John Wiley & Sons, 1992. p. 39-47.

[6] 6. DUNNE, D.W.; JONES, F.M. & DOENHOFF, M.J. - The purification, characterization, serological activity and hepatotoxic properties of two cationic glycoproteins (alfa-1 and omega-1), from S. mansoni eggs. Parasitology, 103: 225-236, 1991.

[7] 7. EVENGARD, B.; HAMMARSTROM, L.; SMITH, C.J.E. et al. - Subclasss distribution and IgE responses after treatment in human schistosomiasis. Clin. exp. Immunol., 73: 383-388, 1988.

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

[9] 9. GENTILINI, M.; PINON, J.M. & NIEL, G. - Immunoélectrodiffusion sur membrane d'ácetate de cellulose. Applications en parasitologie ŕ propos de resultats préliminaires sur 600 essais. Nouv. Presse méd., 1: 1499-1501, 1972.

[10] 10. JASSIM, A.; HASSAN, K. & CATTY, D. - Antibody isotypes in human schistosomiasis mansoni. Paras. Immunol., 9: 627-650, 1987.

[11] 11. KANAMURA-KAWADA, H.Y. - Antígenos de diferentes formas evolutivas do Schistosoma mansoni para diagnóstico e acompanhamento sorológicos de pacientes submetidos ŕ quimioterapia. Săo Paulo, 1985. (Tese de Doutoramento - Instituto de Cięncias Biomédicas da Universidade de Săo Paulo).

[12] 12. KANAMURA, H.Y.; HOSHINO-SHIMIZU, S.; CAMARGO, M.E. et al. - Class specific antibodies and fluorescent staining patterns in acute and chronic forms of schistosomiasis mansoni. Amer. J. trop. Med. Hyg., 28: 242-248, 1979.

[13] 13. KANAMURA, H.Y.; HOSHINO-SHIMIZU, S. & SILVA, L.C. da - Solubilization of antigens of Schistosoma mansoni adult worms for the passive hemagglutination test. Preliminary reports. Rev. Inst. Med. trop. S. Paulo, 23: 92-95, 1981.

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

[15] 15. KATZ, N.; CHAVES, A. & PELLEGRINO, J. - A simple device for quantitative stool thick smear technique in schistosomiasis mansoni. Rev. Inst. Med. trop. S. Paulo, 14: 397-400, 1972.

[16] 16. LOWRY, O.H.; ROSEBROUGH, N.J.; FARR, A.L. et al. - Protein measurement with the Folin phenol reagent. J. biol. Chem., 193: 265-271, 1951.

[17] 17. MADDISON, S.E.; SLEMENDA, S.B.; TSANG, V.C.W. et al. - Serodiagnosis of schistosomiasis mansoni with microsomal adult worm antigen in an enzyme-linked-immunosorbent-assay using a standard curve developed with a reference serum pool. Amer. J. trop. Med. Hyg., 34: 484-494, 1985.

[18] 18. MOTT, K.E. & DIXON, H. - Collaborative study on antigens for immunodiagnosis of schistosomiasis. Bull. Wld. Hlth. Org., 60: 729-753, 1982.

[19] 19. PINON, J.M. & DROPSY, G. - Étude immunologique des parasitoses humaines. Interęt de I'ELIEDA (Enzyme-linked-immuno-electro-diffusion-assay). Path. Biol., 25: 23-27, 1977.

[20] 20. PINON, J.M. & DROPSY, G. - ELIEDA (Enzyme-linked-immuno-electro-diffusion-assay) application of a combined immunoelectrodiffusion and immunoenzyme method to the study of immune response in parasitic infections. J. immunol. Meth., 16: 15-22, 1977.

[21] 21. PINON, J.M. & DROPSY, G. - Étude de l'ímmunité humorale dans la bilharziose humaine. Interęt de I'ELIEDA (Enzyme-linked-immuno-electro-diffusion-assay). Bull. Soc. Path. exot., 69: 238-244, 1977.

[22] 22. PINON, J.M.; GORSE, J.P. & DROPSY, G. - Exploration de l'ímmunité humorale dans L'aspergillose: intéręt de I'ELIEDA (enzyme-linked-immuno-electro-diffusion-assay). Mycopathologia (Den Haag), 60: 115-120, 1977.

[23] 23. PINON, J.M.; SULAHIAN, A. & DROPSY, G. - The use of ELIEDA (enzyme-linked-immuno-electro-diffusion-assay) in the study of humoral antibodies in human schistosomiasis. Trans. roy. Soc. trop. Med. Hyg., 72: 429-495, 1978.

[24] 24. PINON, J.M.; SULAHIAN, A.; REMY, G. et al. - Immunological study of hydatidosis. I. Evaluation of immunoelectrodiffusion tests and enzyme-linked-immuno-electro-diffusion-assay (ELIEDA) in human hydatidosis. Amer. J. trop. Med. Hyg., 28: 318-324, 1979.

[25] 25. PINON, J.M.; THOANNES, H. & GRUSON, N. - An enzyme-linked-immuno-filtration-assay used to compare infant and maternal antibody profiles in toxoplasmosis. J. immunol. Meth., 77: 15-23, 1985.

[26] 26. ROTHMAN, K.J. & BOICE, J.D. - Epidemiologic analysis with a programmable calculator. Boston, Epidemiology Resources Inc., 1982.

[27] 27. RUIZ-TIBEN, E.; HILLYER, G.V.; KNIGHT, W.B. et al. - Intensity of infection with S. mansoni: its relationship to the sensitivity and specificity of Serologic tests. Amer. J. trop. Med. Hyg., 28: 230-236, 1979.

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