UltramicroELISA para la detección de anticuerpos IgM anti M. leprae

Laferte, José; Abreu, Elba G.; Robaina, René; Verez, Vicente

doi:10.1590/S0036-46651991000600010

Resúmenes

La disponibilidad del sistema Ultramicroanalítico (SUMA) y de un antígeno especie-específico del M. leprae obtenido mediante síntesis química, permitió la normalización y validación de un ultramicroELISA para la detección de anticuerpos IgM específicos a esta micobacteria. El análisis de 433 sueros de banco de sangre y 265 sueros usados para validar el método y clasificados en un grupo control de donantes de banco de sangre (100), un grupo de pacientes tuberculosos (50), un grupo de enfermos de lepra (65) y un grupo de contactos de estos enfermos (50), mostró la especificidad del ensayo para evidenciar la infección con el M. leprae. Los resultados obtenidos del estudio adicional de 140 muestras de suero de contactos de enfermos estuvieron estrechamente correlacionados (r = 0,98) con los resultados obtenidos por la técnica de microELISA convencional. La utilización del SUMA no solo permite un notable ahorro de reactivos si no además facilita la lectura, cálculo, validación y almacenamiento automático de los resultados.

UltramicroELISA

The availability of an ultramicroanalitic system (SUMA) and specie-specific antigen of M. leprae obtained by chemical synthesis, have made possible the standardization and validation of an ultramicroELISA assay for detecting specific human IgM antibodies to this mycobacterium. The specificity of this test to demonstrate the infection with M. leprae was corroborated through a screening of 433 blood bank serum samples and other 265 from diferent groups (100, control group, 50 tuberculosis patients, 65 leprosy patients, 50 from household). The results obtained in the aditional study of 140 household sero showed a high correlation (r = 0.98) with the conventional microELISA method. The use of SUMA allows saving reagents and time since sample handling, plate reading, print out and storing the data are computer assisted.

SOROLOGIA

UltramicroELISA para la detección de anticuerpos IgM anti M. leprae

UltramicroELISA assay for the detection of human IgM antibodies to M. leprae

José Laferte^I; Elba G. Abreu^I; René Robaina^II; Vicente Verez^III

^IInstituto de Medicina Tropical "Pedro Kourí" Apdo. 601 Siboney, Ciudad Habana, Cuba

^IICentro de Inmunoensayo. Apdo. 6945. Ciudad Habana, Cuba

^IIILaboratorio de Química de los Carbohidratos. Universidad de la Habana. Facultad de Química. Ciudad Habana, Cuba

^{Dirección para correspondencia} Dirección para correspondencia: Lic. José Laferté Instituto de Medicina Tropical "Pedro Kourí" Apdo. 601 Siboney, Ciudad de la Habana, Cuba

RESUMEN

La disponibilidad del sistema Ultramicroanalítico (SUMA) y de un antígeno especie-específico del M. leprae obtenido mediante síntesis química, permitió la normalización y validación de un ultramicroELISA para la detección de anticuerpos IgM específicos a esta micobacteria. El análisis de 433 sueros de banco de sangre y 265 sueros usados para validar el método y clasificados en un grupo control de donantes de banco de sangre (100), un grupo de pacientes tuberculosos (50), un grupo de enfermos de lepra (65) y un grupo de contactos de estos enfermos (50), mostró la especificidad del ensayo para evidenciar la infección con el M. leprae.

Los resultados obtenidos del estudio adicional de 140 muestras de suero de contactos de enfermos estuvieron estrechamente correlacionados (r = 0,98) con los resultados obtenidos por la técnica de microELISA convencional.

La utilización del SUMA no solo permite un notable ahorro de reactivos si no además facilita la lectura, cálculo, validación y almacenamiento automático de los resultados.

Unitermos: UltramicroELISA

SUMMARY

The availability of an ultramicroanalitic system (SUMA) and specie-specific antigen of M. leprae obtained by chemical synthesis, have made possible the standardization and validation of an ultramicroELISA assay for detecting specific human IgM antibodies to this mycobacterium. The specificity of this test to demonstrate the infection with M. leprae was corroborated through a screening of 433 blood bank serum samples and other 265 from diferent groups (100, control group, 50 tuberculosis patients, 65 leprosy patients, 50 from household). The results obtained in the aditional study of 140 household sero showed a high correlation (r = 0.98) with the conventional microELISA method. The use of SUMA allows saving reagents and time since sample handling, plate reading, print out and storing the data are computer assisted.

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Full text available only in PDF format.

Recebido para publicação em 16/05/1990.

Aceito para publicação em 01/04/1991.

1. AGUADO SANCHEZ, G.; MALIK, A.; TOUGNE, C.; LAMBERT, P.H. & ENGERS, H.D. - Simplification and standarization of serodiagnostic tests for leprosy based on Phenolic Glycolipid-I (GP-I) antigen. Symposium on the immunology of leprosy. Leprosy Rev., 57 (Suppl. 2): 83-93, 1986.
2. AVRAMEAS, J. - Coupling of enzimes to proteins with glutaraldehyde. Immunochemistry, 6: 43-52, 1969.
3. BACH, M.A.; WALLACH, D.; FLAGEUL, B.; HOFFENBACH, A. & COTTENOT, F. - Antibodies to phenolic glycolipid - 1 and to whole Mycobacterium leprae in leprosy patients: evolution during therapy. Int. J. Leprosy, 54: 256-267, 1986.
4. BRETT, S.J.; PAYNE, S.N.; GIGG, J.; BURGESS, P. & GIGG, R. - Use of synthetic glycoconjugates containing the Mycobacterium leprae specific and immunodominant epitope of phenolic glycolipid I in the serology of leprosy. Clin. exp. Immunol., 64: 476-483, 1986.
5. ABE, M.; MINAGAWA, F.; YOSHINO, Y.; OZAWA, T.; SAIKAWA, K. & SAITO, T. - Fluorescent Leprosy Antibody Absorption (FLA-ABS) Test for Detecting Subclinical Infection with Mycobacterium leprae. J. Int. Leprosy, 48: 109-119, 1980.
6. BUCHANAN, T.M.; YOUNG, D.B.; MILLER, R.A. & KHANOLKAR, S.R. - Serodiagnosis of infection with Mycobacterium leprae Int. J. Leprosy, 51: 524-530, 1983.
7. CHO, S.N.; YANAGIHARA, D.L.; HUNTER, S.W.; GELBER, R.H. & BRENNAN, P.J. - Serological specificity of phenolic glycolipid I from Mycobacterium leprae and use in serodiagnosis of leprosy. Infect. Immun., 41: 1077-1083, 1983.
8. CHO, S.N.; FUJIWARA, T.; HUNTER, S.W.; REA, T.H.; GELBER, R.H. & BRENNAN, P.J. - Use of an artificial antigen containing the 3,6-di-O-methyl-B-D-glucopyranosyl epitope for the serodiagnosis of leprosy. J. infect. Dis., 150: 311-322, 1984.
9. FUJIWARA, T.; HUNTER, S.W.; CHO, S.N.; ASPINALL, G.O. & BRENNAN, P.J. - Chemical synthesis and serology of disaccharides and trisaccharides of phenolic glycolipid antigens from the leprosy bacillus and preparation of disaccharide protein conjugate for serodiagnosis of leprosy. Infect. Immun., 43: 245-252, 1984.
10. GIGG, J.; GIGG, R.; PAYNE, S. & CONANT, R. - The allyl group for protection in carbohydrate chemistry. 17* synthesis of propyl-O-(3,6-Di-O-methyl-B-D-glucopyranosyl)-(1 ® 4)-0-(2,3-Di-0-Methyl-µ-L-rhamanopyranosyl)-(1 ® 2)-3-O-methyl-L-rhamnopyranoside: the oligosaccharide portion of the major serologically active glucolipid from Mycobacterium leprae. Chem. Phys. Lip., 38: 299, 1985.
11. HUNTER, S.W. & BRENNAN, P.J. - A novel phenolic glycolipid from Mycobacterium leprae possibly involved in immunogenecity and pathogenicity. J. Bact., 147: 728-735, 1981.
12. HUNTER, S.W.; FUJIWARA, T. & BRENNAN, P.J. - Structure and antigenicity of the major specific glycolipid antigen of Mycobacterium leprae J. biol. Chem., 257: 15072-15078, 1982.
13. MARIŃO, J.R.; BENCOMO, V.V.; GONZALEZ, L. & PEREZ, C.S. - Synthesis of allyl and benzyl 4-0 (3,6 di-O-methyl-8-D) glucopiranosil 1-2, 3-Di-O methyl-L rhamnopyranoside. Carbohydr. Res., 165: 197-206, 1987.
14. SANSONETTI, P.H. & LAGRANGE, P.H. - The immunology of leprosy: speculations on the leprosy spectrum. Rev. infect. Dis., 3: 422-469, 1981.
15. YOUNG, D.B. & BUCHANAN, T.M. - A serological test for leprosy with a glycolipid specific for Mycobacterium leprae. Science, 221: 1057-1059, 1983.
16. YOUNG, D.B.; DISSANAYAKE, S.; MILLER, R.A.; KHANOLKAR, S.R. & BUCHANAN, T.M. - Humans respond predominantly with IgM immunoglobulin to the specie-specific glycolipid of Mycobacterium leprae. J. infect. Dis., 149: 870-873, 1984.
17. YOUNG, D.B.; KHANOLKAR, S.R.; BARG, L.L. & BUCHANAN, T.M. - Generation and characterization of monoclonal antibodies to the phenolic glycolipid of Mycobacterium leprae. Infect. Immun., 43: 183-188, 1984.

Dirección para correspondencia:

Lic. José Laferté

Instituto de Medicina Tropical "Pedro Kourí"

Apdo. 601

Siboney, Ciudad de la Habana, Cuba

Fechas de Publicación

Publicación en esta colección
11 Set 2006
Fecha del número
Dic 1991

Histórico

Recibido
16 Mayo 1990
Acepto
01 Abr 1991

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

[1] 1. AGUADO SANCHEZ, G.; MALIK, A.; TOUGNE, C.; LAMBERT, P.H. & ENGERS, H.D. - Simplification and standarization of serodiagnostic tests for leprosy based on Phenolic Glycolipid-I (GP-I) antigen. Symposium on the immunology of leprosy. Leprosy Rev., 57 (Suppl. 2): 83-93, 1986.

[2] 2. AVRAMEAS, J. - Coupling of enzimes to proteins with glutaraldehyde. Immunochemistry, 6: 43-52, 1969.

[3] 3. BACH, M.A.; WALLACH, D.; FLAGEUL, B.; HOFFENBACH, A. & COTTENOT, F. - Antibodies to phenolic glycolipid - 1 and to whole Mycobacterium leprae in leprosy patients: evolution during therapy. Int. J. Leprosy, 54: 256-267, 1986.

[4] 4. BRETT, S.J.; PAYNE, S.N.; GIGG, J.; BURGESS, P. & GIGG, R. - Use of synthetic glycoconjugates containing the Mycobacterium leprae specific and immunodominant epitope of phenolic glycolipid I in the serology of leprosy. Clin. exp. Immunol., 64: 476-483, 1986.

[5] 5. ABE, M.; MINAGAWA, F.; YOSHINO, Y.; OZAWA, T.; SAIKAWA, K. & SAITO, T. - Fluorescent Leprosy Antibody Absorption (FLA-ABS) Test for Detecting Subclinical Infection with Mycobacterium leprae. J. Int. Leprosy, 48: 109-119, 1980.

[6] 6. BUCHANAN, T.M.; YOUNG, D.B.; MILLER, R.A. & KHANOLKAR, S.R. - Serodiagnosis of infection with Mycobacterium leprae Int. J. Leprosy, 51: 524-530, 1983.

[7] 7. CHO, S.N.; YANAGIHARA, D.L.; HUNTER, S.W.; GELBER, R.H. & BRENNAN, P.J. - Serological specificity of phenolic glycolipid I from Mycobacterium leprae and use in serodiagnosis of leprosy. Infect. Immun., 41: 1077-1083, 1983.

[8] 8. CHO, S.N.; FUJIWARA, T.; HUNTER, S.W.; REA, T.H.; GELBER, R.H. & BRENNAN, P.J. - Use of an artificial antigen containing the 3,6-di-O-methyl-B-D-glucopyranosyl epitope for the serodiagnosis of leprosy. J. infect. Dis., 150: 311-322, 1984.

[9] 9. FUJIWARA, T.; HUNTER, S.W.; CHO, S.N.; ASPINALL, G.O. & BRENNAN, P.J. - Chemical synthesis and serology of disaccharides and trisaccharides of phenolic glycolipid antigens from the leprosy bacillus and preparation of disaccharide protein conjugate for serodiagnosis of leprosy. Infect. Immun., 43: 245-252, 1984.

[10] 10. GIGG, J.; GIGG, R.; PAYNE, S. & CONANT, R. - The allyl group for protection in carbohydrate chemistry. 17* synthesis of propyl-O-(3,6-Di-O-methyl-B-D-glucopyranosyl)-(1 ® 4)-0-(2,3-Di-0-Methyl-µ-L-rhamanopyranosyl)-(1 ® 2)-3-O-methyl-L-rhamnopyranoside: the oligosaccharide portion of the major serologically active glucolipid from Mycobacterium leprae. Chem. Phys. Lip., 38: 299, 1985.

[11] 11. HUNTER, S.W. & BRENNAN, P.J. - A novel phenolic glycolipid from Mycobacterium leprae possibly involved in immunogenecity and pathogenicity. J. Bact., 147: 728-735, 1981.

[12] 12. HUNTER, S.W.; FUJIWARA, T. & BRENNAN, P.J. - Structure and antigenicity of the major specific glycolipid antigen of Mycobacterium leprae J. biol. Chem., 257: 15072-15078, 1982.

[13] 13. MARIŃO, J.R.; BENCOMO, V.V.; GONZALEZ, L. & PEREZ, C.S. - Synthesis of allyl and benzyl 4-0 (3,6 di-O-methyl-8-D) glucopiranosil 1-2, 3-Di-O methyl-L rhamnopyranoside. Carbohydr. Res., 165: 197-206, 1987.

[14] 14. SANSONETTI, P.H. & LAGRANGE, P.H. - The immunology of leprosy: speculations on the leprosy spectrum. Rev. infect. Dis., 3: 422-469, 1981.

[15] 15. YOUNG, D.B. & BUCHANAN, T.M. - A serological test for leprosy with a glycolipid specific for Mycobacterium leprae. Science, 221: 1057-1059, 1983.

[16] 16. YOUNG, D.B.; DISSANAYAKE, S.; MILLER, R.A.; KHANOLKAR, S.R. & BUCHANAN, T.M. - Humans respond predominantly with IgM immunoglobulin to the specie-specific glycolipid of Mycobacterium leprae. J. infect. Dis., 149: 870-873, 1984.

[17] 17. YOUNG, D.B.; KHANOLKAR, S.R.; BARG, L.L. & BUCHANAN, T.M. - Generation and characterization of monoclonal antibodies to the phenolic glycolipid of Mycobacterium leprae. Infect. Immun., 43: 183-188, 1984.

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