UltramicroELISA para la deteccion de anticuerpos IgM al Mycobacterium leprae utilizando muestras de sangre seca

Torrella, Adriana; Solis, Rosa L.; Rodriguez, Nerys; Medina, Yadira; Pita, Martha; Perez, Ivis; Licourt, Nubia

doi:10.1590/S0036-46651994000200007

Resúmenes

En este trabajo se establecen las condiciones optimas para la detección de anticuerpos IgM al glicolipido fenólico-I (GF-I) en muestras de sangre en papel de filtro utilizando el UltramicroELISA HANSEN y la tecnologia SUMA. Se estudiaron 300 donantes de sangre y 58 pacientes leprosos. Para estas dos poblaciones se compararon los resultados de muestras de sangre seca colectadas en papel de filtro SS-2992 con los de suero, y se obtuvo una correlación de 0.919 para donantes de sangre, 0.969 para pacientes y 0.954 para el total de las dos poblaciones. Se obtuvo una coincidencia de 98% en pacientes y 96% en donantes. En la población de pacientes la sensibilidad fue de 93% y la especificidad de 100% para las muestras de sangre seca evaluadas por el UMELISA HANSEN, con respecto a las muestras de suero analizadas por esta misma tecnica.

Lepra; IgM anti-glicolipido fenolico-I (IgM ANTI GF-I) UltramicroELISA; Sangre seca en papel de filtro

In this work the adequate conditions for the detection of IgM antibodies to PGL-I in eluates from dried blood spots using the UMELISA HANSEN and the SUMA technology are established. A total of 300 blood donors and 58 leprosy patients samples were studied. For both populations, in the previously established conditions, we compared the results of the eluates from dried blood spots with the serum samples, and was obtained a correlation of 0.919 in blood donors, 0.969 in patients, and 0.954 for the total of both populations. It was also obtained a level of agreemnet of 98% in patients and 96% in blood donors. In the patients population was found a sensitivity of 93% and an specificity of 100% for the eluates of dried blood spots evaluated by the UMELISA HANSEN in the comparison with the serum samples analized by the same assay.

SOROLOGIA

UltramicroELISA para la deteccion de anticuerpos IgM al Mycobacterium leprae utilizando muestras de sangre seca

UltramicroELISA assay for the detection of IgM antibodies to Mycobacterium leprae using eluates of dried blood spots

Adriana Torrella^I; Rosa L. Solis^I; Nerys Rodriguez^I; Yadira Medina^II; Martha Pita^II; Ivis Perez^II; Nubia Licourt^II

^ICentro de Inmunoensayo. Apart 6945. Ciudad Habana. CUBA

^IIHospital Especializado Dermatológico: G. Fdez. Hdez-Baquero Rincón, Habana, CUBA

^{Dirección para correspondencia} Dirección para correspondencia: Lic. Adriana Torrella Centro de Inmunoensayo Apart 6945. Calle 134 y Ave 25 Playa, Ciudad Habana, Cuba

RESUMEN

En este trabajo se establecen las condiciones optimas para la detección de anticuerpos IgM al glicolipido fenólico-I (GF-I) en muestras de sangre en papel de filtro utilizando el UltramicroELISA HANSEN y la tecnologia SUMA. Se estudiaron 300 donantes de sangre y 58 pacientes leprosos. Para estas dos poblaciones se compararon los resultados de muestras de sangre seca colectadas en papel de filtro SS-2992 con los de suero, y se obtuvo una correlación de 0.919 para donantes de sangre, 0.969 para pacientes y 0.954 para el total de las dos poblaciones. Se obtuvo una coincidencia de 98% en pacientes y 96% en donantes. En la población de pacientes la sensibilidad fue de 93% y la especificidad de 100% para las muestras de sangre seca evaluadas por el UMELISA HANSEN, con respecto a las muestras de suero analizadas por esta misma tecnica.

Unitermos: Lepra; IgM anti-glicolipido fenolico-I (IgM ANTI GF-I) UltramicroELISA; Sangre seca en papel de filtro.

SUMMARY

In this work the adequate conditions for the detection of IgM antibodies to PGL-I in eluates from dried blood spots using the UMELISA HANSEN and the SUMA technology are established. A total of 300 blood donors and 58 leprosy patients samples were studied. For both populations, in the previously established conditions, we compared the results of the eluates from dried blood spots with the serum samples, and was obtained a correlation of 0.919 in blood donors, 0.969 in patients, and 0.954 for the total of both populations. It was also obtained a level of agreemnet of 98% in patients and 96% in blood donors. In the patients population was found a sensitivity of 93% and an specificity of 100% for the eluates of dried blood spots evaluated by the UMELISA HANSEN in the comparison with the serum samples analized by the same assay.

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Full text available only in PDF format.

Recebido para publicação em 01/07/1993.

Aceito para publicação em 09/11/1993.

1. BAUMGART, K.; BRITTON, W.; BASTEN, A. & BAGSHAWE, A. - Use of phenolic glycolipid for serodiagnosis of leprosy in a high prevalence village in Papua, New Guinea. Trans. roy. Soc. trop. Med. Hyg., 81: 1030, 1987.
2. BUCHANAN, T.M.; YOUNG, D.B.; MILLER, R.A. & KHANOLKAR, S.R. - Serodiagnosis of infection with Mycobac-terium leprae. Int. J. Leprosy, 51: 524, 1983.
3. CASSOL, S.; SALAS, T.; ARELLA, M. & NEUMANN, P. - Use of dried blood spots specimens in the detection of human immunodeficiency virus type 1 by the polymerase chain reaction. J. clin. Microbiol., 29: 667-671, 1991.
4. CHUJOR, S.N.; BERNHEIMER, H.; LEVIS, R.W. & SCHWERER, B. - Serum IgA and IgM antibodies against Mycobacterium leprae derived PGL-I: a comparative study in leprosy patients and their contacts. Int. J. Leprosy, 59: 441-449, 1991.
5. DHANDAYUTHAPANI, S.; ANANDAN, D.; VASANTHI, B. & BHATIA, V.N. - Use of eluates of filter paper blood spots in ELISA for the serodiagnosis of leprosy. Indian J. med. Res., 89: 150, 1989.
6. DHANDAYUTHAPANI, S.; ANANDAN, D. & BHATIA, V.N. - ELISA & lepromin skin tests in household contacts of leprosy patients. Indian. J. med. Res., 91: 431-436, 1990.
7. DISSANAYAKE, S.; YOUNG, D.B.; KHANOLKAR, S.R.; MILLER, R.A. & BUCHANAN, T.M. - Evaluation of the significance of antibodies to phenolic glycolipid of M. leprae in leprosy patients and their contacts. Int. J. Leprosy, 52 (Suppl. 4): 691, 1984.
8. GONZALEZ, A.B.; GLEZ-ABREU, E. & VALDEZ-PORTELA, A. - Leprosy: a brief account on the subject. Rev. Med. trop. (Cuba), 40: 67-81, 1988.
9. GUIMARĂES, M.C.S.; CASTILHO, E.A. & CELESTE, B.J. - Long-term storage of IgG and IgM on filter paper for use in parasitic disease seroepidemiology surveys. Bull. Pan Amer. Hith. Org., 19: 16-28, 1985.
10. HUSSAIN, R.; JAMIL, S. & KIFAYET, A. - Quantitation of IgM antibodies to the M. leprae synthetic disaccharide can predict early bacterial multiplication in leprosy. Int. J. Leprosy, 58: 491-502, 1990.
11. MEEKER, C.H.; SCHULLER-LEVIS, G. & FUSCO, F. - Sequential monitoring of leprosy patients with serum antibody levels to PGL-I, a synthetic analog of PGL-I, and Mycobacterial lipoarabinomannan. Int. J. Leprosy, 58: 503-511, 1990.
12. MOUNT, L.D. & CHURCHILL, C.F. - Determination of mefloquine in blood, filter paper adsorbed blood and urine by 9 fluorenylmethyl chloroformate derivatization followed by liquid chromatography with fluorescence detection. J. Chromatogr., 564: 181-193, 1991.
13. MOUDGIL, K.D.; MISHRA R. & TALWAR, G.P. - Comparative evaluation of enzyme immunoassays based on synthetic glyco-conjugates and PGL-I for immunodiagnosis of leprosy. Indian J. Leprosy, 62: 60-65, 1990.
14. SHRIPAD, A.P.; GOPAL, R.M.D. & SINHA, S. - Screening of anti-M. leprae antibodies in the blood samples eluted from filter paper blood blots. Int. J. Leprosy, 58: 123-126, 1990.
15. VARNIER, O.E.; LILLO, F.B.; REINA, S. & TERRAGNA, A. - Whole blood collection on filter paper is an effective means of obtaining samples for human immunodeficiency virus antibody assay. AIDS Res. Hum. Retroviruses, 4: 131-136, 1988.
16. ZICKER, F.; SMITH, P.G.; LUQUETTI, A.O. & OLIVEIRA, O.S. - Mass screening for Trypanosoma cruzi infections using the immunofluorescence, ELISA and haemagglutination tests on serum samples and on blood eluates from filter paper. Bull. Wld. Hith. Org., 68: 465-471, 1990.

Dirección para correspondencia:

Lic. Adriana Torrella

Centro de Inmunoensayo

Apart 6945. Calle 134 y Ave 25

Playa, Ciudad Habana, Cuba

Fechas de Publicación

Publicación en esta colección
20 Set 2006
Fecha del número
Abr 1994

Histórico

Recibido
01 Jul 1993
Acepto
09 Nov 1993

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

[1] 1. BAUMGART, K.; BRITTON, W.; BASTEN, A. & BAGSHAWE, A. - Use of phenolic glycolipid for serodiagnosis of leprosy in a high prevalence village in Papua, New Guinea. Trans. roy. Soc. trop. Med. Hyg., 81: 1030, 1987.

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

[3] 3. CASSOL, S.; SALAS, T.; ARELLA, M. & NEUMANN, P. - Use of dried blood spots specimens in the detection of human immunodeficiency virus type 1 by the polymerase chain reaction. J. clin. Microbiol., 29: 667-671, 1991.

[4] 4. CHUJOR, S.N.; BERNHEIMER, H.; LEVIS, R.W. & SCHWERER, B. - Serum IgA and IgM antibodies against Mycobacterium leprae derived PGL-I: a comparative study in leprosy patients and their contacts. Int. J. Leprosy, 59: 441-449, 1991.

[5] 5. DHANDAYUTHAPANI, S.; ANANDAN, D.; VASANTHI, B. & BHATIA, V.N. - Use of eluates of filter paper blood spots in ELISA for the serodiagnosis of leprosy. Indian J. med. Res., 89: 150, 1989.

[6] 6. DHANDAYUTHAPANI, S.; ANANDAN, D. & BHATIA, V.N. - ELISA & lepromin skin tests in household contacts of leprosy patients. Indian. J. med. Res., 91: 431-436, 1990.

[7] 7. DISSANAYAKE, S.; YOUNG, D.B.; KHANOLKAR, S.R.; MILLER, R.A. & BUCHANAN, T.M. - Evaluation of the significance of antibodies to phenolic glycolipid of M. leprae in leprosy patients and their contacts. Int. J. Leprosy, 52 (Suppl. 4): 691, 1984.

[8] 8. GONZALEZ, A.B.; GLEZ-ABREU, E. & VALDEZ-PORTELA, A. - Leprosy: a brief account on the subject. Rev. Med. trop. (Cuba), 40: 67-81, 1988.

[9] 9. GUIMARĂES, M.C.S.; CASTILHO, E.A. & CELESTE, B.J. - Long-term storage of IgG and IgM on filter paper for use in parasitic disease seroepidemiology surveys. Bull. Pan Amer. Hith. Org., 19: 16-28, 1985.

[10] 10. HUSSAIN, R.; JAMIL, S. & KIFAYET, A. - Quantitation of IgM antibodies to the M. leprae synthetic disaccharide can predict early bacterial multiplication in leprosy. Int. J. Leprosy, 58: 491-502, 1990.

[11] 11. MEEKER, C.H.; SCHULLER-LEVIS, G. & FUSCO, F. - Sequential monitoring of leprosy patients with serum antibody levels to PGL-I, a synthetic analog of PGL-I, and Mycobacterial lipoarabinomannan. Int. J. Leprosy, 58: 503-511, 1990.

[12] 12. MOUNT, L.D. & CHURCHILL, C.F. - Determination of mefloquine in blood, filter paper adsorbed blood and urine by 9 fluorenylmethyl chloroformate derivatization followed by liquid chromatography with fluorescence detection. J. Chromatogr., 564: 181-193, 1991.

[13] 13. MOUDGIL, K.D.; MISHRA R. & TALWAR, G.P. - Comparative evaluation of enzyme immunoassays based on synthetic glyco-conjugates and PGL-I for immunodiagnosis of leprosy. Indian J. Leprosy, 62: 60-65, 1990.

[14] 14. SHRIPAD, A.P.; GOPAL, R.M.D. & SINHA, S. - Screening of anti-M. leprae antibodies in the blood samples eluted from filter paper blood blots. Int. J. Leprosy, 58: 123-126, 1990.

[15] 15. VARNIER, O.E.; LILLO, F.B.; REINA, S. & TERRAGNA, A. - Whole blood collection on filter paper is an effective means of obtaining samples for human immunodeficiency virus antibody assay. AIDS Res. Hum. Retroviruses, 4: 131-136, 1988.

[16] 16. ZICKER, F.; SMITH, P.G.; LUQUETTI, A.O. & OLIVEIRA, O.S. - Mass screening for Trypanosoma cruzi infections using the immunofluorescence, ELISA and haemagglutination tests on serum samples and on blood eluates from filter paper. Bull. Wld. Hith. Org., 68: 465-471, 1990.

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