El inmunoensayo enzimático (ELISA) en el diagnóstico serológico de Plasmodium vivax

Perez, Hilda A.; De La Rosa, Mercedes

doi:10.1590/S0036-46651990000300008

Resúmenes

Comunicamos nuestra experiencia con la aplicación del inmunoensayo enzimático al diagnóstico serológico de Plasmodium vivax con antígeno homólogo. Este se obtuvo a partir de una muestra de sangre de un paciente y luego de lisar los glóbulos rojos con detergente (NP-40). En un "pool" conformado con 11 sueros de pacientes con malaria por P. vivax se encontraron anticuerpos de las clases IgM, IgG e IgA. Se introdujo además la proteína A como sonda secundaria para determinar la respuesta anti-P. vivax en una muestra de 30 sueros de pacientes con un primer episodio palúdico. Se encontró un 93% de correlación con el diagnóstico parasitológico y la prueba resultó específica y reproducible.

ELISA was evaluated for the serodiagnosis of Plasmodium vivax using homologous antigen. This was a crude fraction obtained after detergent (NP-40) lysis of human parasitized red blood cells. Antibodies of the classes IgM, IgG, IgA were determined in a pool of eleven sera from patients with P. vivax malaria. The protein A was introduced as secondary probe to screen P. vivax antibodies in 30 sera of patients harbouring a first episode of P. vivax malaria. There was a correlation of 93% with the parasitological diagnosis and the test resulted specific and reproducible.

SOROLOGIA

El inmunoensayo enzimático (ELISA) en el diagnóstico serológico de Plasmodium vivax

The immunoenzymatic assay (ELISA) in the serodiagnosis of Plasmodium vivax

Hilda A. Perez; Mercedes De La Rosa

Instituto Venezolano de Investigaciones Científicas, IVIC, Centro de Microbiología y Biología Celular, Laboratório de Inmunoparasitología. Apdo. 21827, Caracas 1020A Venezuela

RESUMEN

Comunicamos nuestra experiencia con la aplicación del inmunoensayo enzimático al diagnóstico serológico de Plasmodium vivax con antígeno homólogo. Este se obtuvo a partir de una muestra de sangre de un paciente y luego de lisar los glóbulos rojos con detergente (NP-40). En un "pool" conformado con 11 sueros de pacientes con malaria por P. vivax se encontraron anticuerpos de las clases IgM, IgG e IgA. Se introdujo además la proteína A como sonda secundaria para determinar la respuesta anti-P. vivax en una muestra de 30 sueros de pacientes con un primer episodio palúdico. Se encontró un 93% de correlación con el diagnóstico parasitológico y la prueba resultó específica y reproducible.

Unitermos: Malaria; P. vivax; Anticuerpos; ELISA.

SUMMARY

ELISA was evaluated for the serodiagnosis of Plasmodium vivax using homologous antigen. This was a crude fraction obtained after detergent (NP-40) lysis of human parasitized red blood cells. Antibodies of the classes IgM, IgG, IgA were determined in a pool of eleven sera from patients with P. vivax malaria. The protein A was introduced as secondary probe to screen P. vivax antibodies in 30 sera of patients harbouring a first episode of P. vivax malaria. There was a correlation of 93% with the parasitological diagnosis and the test resulted specific and reproducible.

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AGRADECIMIENTO

Se agradece la colaboración técnica del Lic. José Bolívar y el constante apoyo de la Dirección de Malariología y Saneamiento Ambiental del MSAS. Vaya también nuestro agradecimiento al Banco Municipal de Sangre de Caracas, especialmente a la Dra. Luz Nuñez por su desinteresada colaboración en la donación de las muestras controles.

Financiado por OPS/DRS/RG Ven 86/126 y CONICIT S1-1849.

Recebido para publicação em 13/7/1989.

1. Al KAISSI, S. & MOSTRATOS, A. Assessment of substrates for horseradish peroxidase in enzyme immunoassay. J. immunol. Meth., 58: 127-132, 1987.
2. AVRAHAM, H.; GOLENSER, J.; SPIRA, D.T. & SULITZEANU, D. Plasmodium falciparum: assay of antigens and antibodies by means of a solid phase radioimmunoassay with radioiodinated staphylococcal protein A. Trans roy. Soc. trop. Med. Hyg., 75: 421-425, 1981.
3. BARNWELL, J.W. Antigens of Plasmodium vivax blood stage parasites identified by monoclonal antibodies. Mem. Inst. Oswaldo Cruz, 81 (Suppl. 2): 59-61, 1986.
4. BERZINS, K.; WAHLGREN, M. & PERLMANN, P. Studies on the specificity of anti-erythrocyte antibodies in the serum of patients with malaria. Clin. exp. Immunol., 54: 313-318, 1983.
5. BERZINS, K.; PERLMANN, H.; WAHLIN, B.; RUANGJI-RACHUPORN, W.; HOGH, B.; PETERSEN, E.; BJORKMAN, A. & PERLMANN, P. Antibodies to repeated amino acid sequence in Pfl55, a merozoite associated antigen of Plasmodium falciparum. Mem. Inst. Oswaldo Cruz, 81 (Suppl. 2): 77-81, 1986.
6. BRADFOR, O.F. Adaptation of the Bradford-protein assay to membrane-bound proteins by solubilizing in glucopyranoside detergents. Analyt. Biochem., 162: 11-17, 1987.
7. BRUCE-CHWATT, L.J.; DRAPER, C.C. & KONFORTION, P. Seroepidemiological evidence of erradication of malaria from Mauritius. Lancet, 2: 547-551, 1973.
8. BRUCE-CHWATT, L.J.; DRAPER, C.C.; AVRAMIDIS, D. & KAZANDZOGLOU, O. Seroepidemiological surveillance of disappearing malaria in Greece. J. trop. Med. Hyg., 78: 194-200, 1975.
9. CASTRO, J.; BORGES, L.; OTERO, M.A. & GUERRERO, L. Malaria. In: PEREZ, H. & HERNANDEZ, A. Seminario sobre enfermedades tropicales en Venezuela, 1., Caracas, MCT/MSAS/OPS., 1985. p. 105-139.
10. CORNILLE-BROGGER, R.; MATHEWS, H.M.; STOREY, J.; ASHKAR, T.S.; BROGGER, S. & MOLINEAUX, L. Changing patterns in the humoral immune response to malaria before, during and after the application of control measures: a longitudinal study in the West African savanna. Bull. Wld. Hlth. Org., 56: 579-600, 1978.
11. GABALDON, A.; BERTI, A.L.; GUERRERO, L. & GARCIA MARTIN, G. Erradicación de la malaria en Venezuela. Programa evolución y estado actual. Rev. venez. Sanid., (Supl. al No. 3): 290-336, 1961.
12. GABALDON, A. What can and cannot be achieved with conventional anti-malaria measures. Amer. J. trop. Med. Hyg., 27: 653-658, 1978.
13. GREENWOOD, B.M. The impact of malaria chemoprophylaxis on the immune status of Africans. Bull. Wld. Hlth. Org., 62 (suppl): 69-75, 1984.
14. KRONVALL, G. & WILLIAMS JR., R.C. Differences in antiprotein A activity among IgG subgroups. J. Immunol., 103: 828-833, 1969.
15. MACKEY, L.J.; McGREGOR, I.A.; PAOUNOVA, N. & LAMBERT, P.H. Diagnosis of Plasmodium falciparum infection in man: detection of parasite antigens by ELISA. Bull. Wld. Hlth. Org., 60: 69-75, 1982.
16. NOTANI, G.W.; PARSONS, J.A. & ERLANDSEN, S.L. Versatility of Staphylococcus aureus Protein A in immunocytochemistry. J. Histochem. Cytochem., 27: 1438-1444, 1979.
17. QUAKYI, I.A. The development and validation of an enzyme linked immunosorbent assay for malaria. Tropenmed. Parasitol., 31: 325-333, 1980.
18. ROMANO, E. & ROMANO, M. Staphylococcal protein A bound to coloidal gold a useful reagent to label antigen-antibody sites in electron microscopy. Immunochemistry, 14: 711-715, 1977.
19. SHERMAN, I.W. Membrane structure and function of malaria parasites and the infected erythrocyte. Parasitology, 91: 609-645, 1985.
20. TANDON, A.; SAXENA, R.P.; BHATIA, B. & SAXENA, K.C. The enzyme linked immunosorbent assay in the immunodiagnosis of human malaria. Trans. roy. Soc. trop. Med. Hyg., 76: 371-372, 1982.
21. TARGETT, G.A.T. Antibody response to Plasmodium falciparum malaria. Comparison of immunoglobulin concentrations, antibody titres and the antigenicity of different asexual forms of the parasite. Clin. exp. Immunol., 7: 501-517, 1970.
22. VOLLER, A.; BIDWELL, D.; HULDT, G. & ENGVALL, E. A microplate method of enzyme linked immunosorbent assay and its application to malaria. Bull. Wld. Hlth. Org., 51: 209-211, 1974.
23. VOLLER, A.; CORNILLE-BROGGER, R.; STOREY, J. & MOLINEAUX, L. A longitudinal study of Plasmodium falciparum malaria in the West African savanna using the ELISA technique. Bull. Wld. Hlth. Org., 58: 429-438, 1980.
24. WAHLGREN, M.; BERZINS, K.; PERLMANN, P. & PERSSON, M. Characterization of the humoral immune response in Plasmodium falciparum malaria. II. IgG subclass levels of anti-P. falciparum antibodies in different sera. Clin. exp. Immunol., 54: 135-142, 1983.
25. ZOUALI, M.; DRUILHE, P.; GENTILINI, M. & EYQUEM, A. High titres of anti-T antibodies and other haemagglutinins in human malaria. Clin. exp. Immunol., 50: 83-91, 1982.

Fechas de Publicación

Publicación en esta colección
13 Set 2006
Fecha del número
Jun 1990

Histórico

Recibido
13 Jul 1989

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

[1] 1. Al KAISSI, S. & MOSTRATOS, A. Assessment of substrates for horseradish peroxidase in enzyme immunoassay. J. immunol. Meth., 58: 127-132, 1987.

[2] 2. AVRAHAM, H.; GOLENSER, J.; SPIRA, D.T. & SULITZEANU, D. Plasmodium falciparum: assay of antigens and antibodies by means of a solid phase radioimmunoassay with radioiodinated staphylococcal protein A. Trans roy. Soc. trop. Med. Hyg., 75: 421-425, 1981.

[3] 3. BARNWELL, J.W. Antigens of Plasmodium vivax blood stage parasites identified by monoclonal antibodies. Mem. Inst. Oswaldo Cruz, 81 (Suppl. 2): 59-61, 1986.

[4] 4. BERZINS, K.; WAHLGREN, M. & PERLMANN, P. Studies on the specificity of anti-erythrocyte antibodies in the serum of patients with malaria. Clin. exp. Immunol., 54: 313-318, 1983.

[5] 5. BERZINS, K.; PERLMANN, H.; WAHLIN, B.; RUANGJI-RACHUPORN, W.; HOGH, B.; PETERSEN, E.; BJORKMAN, A. & PERLMANN, P. Antibodies to repeated amino acid sequence in Pfl55, a merozoite associated antigen of Plasmodium falciparum. Mem. Inst. Oswaldo Cruz, 81 (Suppl. 2): 77-81, 1986.

[6] 6. BRADFOR, O.F. Adaptation of the Bradford-protein assay to membrane-bound proteins by solubilizing in glucopyranoside detergents. Analyt. Biochem., 162: 11-17, 1987.

[7] 7. BRUCE-CHWATT, L.J.; DRAPER, C.C. & KONFORTION, P. Seroepidemiological evidence of erradication of malaria from Mauritius. Lancet, 2: 547-551, 1973.

[8] 8. BRUCE-CHWATT, L.J.; DRAPER, C.C.; AVRAMIDIS, D. & KAZANDZOGLOU, O. Seroepidemiological surveillance of disappearing malaria in Greece. J. trop. Med. Hyg., 78: 194-200, 1975.

[9] 9. CASTRO, J.; BORGES, L.; OTERO, M.A. & GUERRERO, L. Malaria. In: PEREZ, H. & HERNANDEZ, A. Seminario sobre enfermedades tropicales en Venezuela, 1., Caracas, MCT/MSAS/OPS., 1985. p. 105-139.

[10] 10. CORNILLE-BROGGER, R.; MATHEWS, H.M.; STOREY, J.; ASHKAR, T.S.; BROGGER, S. & MOLINEAUX, L. Changing patterns in the humoral immune response to malaria before, during and after the application of control measures: a longitudinal study in the West African savanna. Bull. Wld. Hlth. Org., 56: 579-600, 1978.

[11] 11. GABALDON, A.; BERTI, A.L.; GUERRERO, L. & GARCIA MARTIN, G. Erradicación de la malaria en Venezuela. Programa evolución y estado actual. Rev. venez. Sanid., (Supl. al No. 3): 290-336, 1961.

[12] 12. GABALDON, A. What can and cannot be achieved with conventional anti-malaria measures. Amer. J. trop. Med. Hyg., 27: 653-658, 1978.

[13] 13. GREENWOOD, B.M. The impact of malaria chemoprophylaxis on the immune status of Africans. Bull. Wld. Hlth. Org., 62 (suppl): 69-75, 1984.

[14] 14. KRONVALL, G. & WILLIAMS JR., R.C. Differences in antiprotein A activity among IgG subgroups. J. Immunol., 103: 828-833, 1969.

[15] 15. MACKEY, L.J.; McGREGOR, I.A.; PAOUNOVA, N. & LAMBERT, P.H. Diagnosis of Plasmodium falciparum infection in man: detection of parasite antigens by ELISA. Bull. Wld. Hlth. Org., 60: 69-75, 1982.

[16] 16. NOTANI, G.W.; PARSONS, J.A. & ERLANDSEN, S.L. Versatility of Staphylococcus aureus Protein A in immunocytochemistry. J. Histochem. Cytochem., 27: 1438-1444, 1979.

[17] 17. QUAKYI, I.A. The development and validation of an enzyme linked immunosorbent assay for malaria. Tropenmed. Parasitol., 31: 325-333, 1980.

[18] 18. ROMANO, E. & ROMANO, M. Staphylococcal protein A bound to coloidal gold a useful reagent to label antigen-antibody sites in electron microscopy. Immunochemistry, 14: 711-715, 1977.

[19] 19. SHERMAN, I.W. Membrane structure and function of malaria parasites and the infected erythrocyte. Parasitology, 91: 609-645, 1985.

[20] 20. TANDON, A.; SAXENA, R.P.; BHATIA, B. & SAXENA, K.C. The enzyme linked immunosorbent assay in the immunodiagnosis of human malaria. Trans. roy. Soc. trop. Med. Hyg., 76: 371-372, 1982.

[21] 21. TARGETT, G.A.T. Antibody response to Plasmodium falciparum malaria. Comparison of immunoglobulin concentrations, antibody titres and the antigenicity of different asexual forms of the parasite. Clin. exp. Immunol., 7: 501-517, 1970.

[22] 22. VOLLER, A.; BIDWELL, D.; HULDT, G. & ENGVALL, E. A microplate method of enzyme linked immunosorbent assay and its application to malaria. Bull. Wld. Hlth. Org., 51: 209-211, 1974.

[23] 23. VOLLER, A.; CORNILLE-BROGGER, R.; STOREY, J. & MOLINEAUX, L. A longitudinal study of Plasmodium falciparum malaria in the West African savanna using the ELISA technique. Bull. Wld. Hlth. Org., 58: 429-438, 1980.

[24] 24. WAHLGREN, M.; BERZINS, K.; PERLMANN, P. & PERSSON, M. Characterization of the humoral immune response in Plasmodium falciparum malaria. II. IgG subclass levels of anti-P. falciparum antibodies in different sera. Clin. exp. Immunol., 54: 135-142, 1983.

[25] 25. ZOUALI, M.; DRUILHE, P.; GENTILINI, M. & EYQUEM, A. High titres of anti-T antibodies and other haemagglutinins in human malaria. Clin. exp. Immunol., 50: 83-91, 1982.

Brasil

Brasil

El inmunoensayo enzimático (ELISA) en el diagnóstico serológico de Plasmodium vivax

The immunoenzymatic assay (ELISA) in the serodiagnosis of Plasmodium vivax

Resúmenes

Fechas de Publicación

Histórico