Deteccion de citomegalovirus mediante la tecnica de inmunoperoxidasa y aislamiento viral

Alvarez, Maritza; Marrero, Miguel; Valdivia, Angel; Garcia, Serafina; Valdes, Odalys; Morier, Luis

doi:10.1590/S0036-46651991000300002

Resúmenes

En el presente estudio se comparó la técnica de inmunoperoxidasa para la detección de citomegalovirus (IPCMV) utilizando anticuerpos monoclonales que reconocen proteínas precoces virales con el método convencional de aislamiento viral en fibroblastos humanos. Un total de 150 muestras de orina fueron examinadas encontrando una sensibilidad de un 89.8% y una especificidad de 91.3% de la técnica de IPCMV comparada con el aislamiento viral. Una de las ventajas que presentó la IPCMV fue la rapidez con que fueron obtenidos los resultados (48 horas) mientras que el aislamiento viral fue como promedio 14 días.

Citomegalovirus; Inmunoperoxidasa; Aislamiento viral

An Immunoperoxidase assay was applied to detect early antigens of Cytomegalovirus (CMV) in 150 urine samples from immunocompromised patients, using the commercial available monoclonal antibody against CMV El3. The detection of early antigen by IP (IPCMV) is compared to the conventional cell culture isolation regarding specificity and sensitivity in order to evaluate is usefulness in the diagnostic of CMV infections. The IPCMV showed a sensitivity of 89.8% and a specificity of 91.3% when compared to the isolation method. The great advantage of the IPCMV is based on the shorter time results are achieved, since 48-72 Hs can be enough to provide evidence of CMV infection, while in the isolation technique cytopatho-genic effect was present around 14 days after sample inoculation.

ARTIGOS ORIGINAIS

Deteccion de citomegalovirus mediante la tecnica de inmunoperoxidasa y aislamiento viral

Cytomegalovirus detection by Immunoperoxidase assay and viral isolation

Maritza Alvarez; Miguel Marrero; Angel Valdivia; Serafina Garcia; Odalys Valdes; Luis Morier

Departamento de Virología. Instituto de Medicina Tropical "Pedro Kourí", Siboney, Ciudad Habana, Cuba

^{Dirección para correspondencia} Dirección para correspondencia: Dra. M. Alvarez Instituto de Medicina Tropical "Pedro Kourí" Apartado Postal Nº 601 Marianao, C. Habana, Cuba

RESUMEN

En el presente estudio se comparó la técnica de inmunoperoxidasa para la detección de citomegalovirus (IPCMV) utilizando anticuerpos monoclonales que reconocen proteínas precoces virales con el método convencional de aislamiento viral en fibroblastos humanos. Un total de 150 muestras de orina fueron examinadas encontrando una sensibilidad de un 89.8% y una especificidad de 91.3% de la técnica de IPCMV comparada con el aislamiento viral. Una de las ventajas que presentó la IPCMV fue la rapidez con que fueron obtenidos los resultados (48 horas) mientras que el aislamiento viral fue como promedio 14 días.

Unitermos: Citomegalovirus; Inmunoperoxidasa; Aislamiento viral.

SUMMARY

An Immunoperoxidase assay was applied to detect early antigens of Cytomegalovirus (CMV) in 150 urine samples from immunocompromised patients, using the commercial available monoclonal antibody against CMV El3. The detection of early antigen by IP (IPCMV) is compared to the conventional cell culture isolation regarding specificity and sensitivity in order to evaluate is usefulness in the diagnostic of CMV infections. The IPCMV showed a sensitivity of 89.8% and a specificity of 91.3% when compared to the isolation method. The great advantage of the IPCMV is based on the shorter time results are achieved, since 48-72 Hs can be enough to provide evidence of CMV infection, while in the isolation technique cytopatho-genic effect was present around 14 days after sample inoculation.

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Full text available only in PDF format.

Recebido para publicação em 16/05/1990

Aceito para publicação em 18/03/1991

1. ALPERT, G.; MAZERON M.C.; COLIMON, R. & PLOTKIN, S. - Rapid detection of human cytomegalovirus in the urine of humans. J. infect. Dis., 152: 631-633, 1985.
2. ANDERSON, C.H. & MICHAELS, R.H. - Cytomegalovirus infections detection by direct fluorescent-antibody technique. Lancet, 2: 308-309, 1987.
3. BLASER, J.M. & COHN, D.L. - Opportunistic infections in patients with AIDS. Clues to the epidemiology of AIDS and the relative virulence of pathogens. J. infect. Dis., 8: 21-30, 1986.
4. COREY, L.; DRAVAGON, J. & BENJAMIN, D. - Rapid methods and automation in microbiology. In: TILTON, R.C., ed - Immunoperoxidase stainning in the clinical virology laboratory, Washington, American Society for Microbiology. p.246-253.
5. DEMMLER, G.J.; BUFFONE, G.J.; SCHIMBOR, C.M. & MAY, R.A. - Detection of cytomegalovirus in urine from newborns by using polymerase chain reaction DNA amplification. J. infect. Dis., 158: 1177-1184, 1988.
6. FORBES, B.A. - Acquisition of cytomegalovirus infections. An update. Clin. Microbiol. Rev., 2: 202-261, 1989.
7. GLEAVES, C.A.; SMITH, T.F.; SHUSTER, E.A. & PEARSON, G.R. - Rapid detection of cytomegalovirus in MRC 5 cells inoculated with urine specimens by using low-speed centrifugation and monoclonal antibody to an early antigen. J. clin. Microbiol., 19: 917-919, 1984.
8. GLENN, J. - Cytomegalovirus infections following renal transplantation. Rev. infect. Dis., 3: 1151-1178, 1981.
9. GOLDSTEIN, L.C.; DOUGALL, J.K.; HACKMAN, R.C.; MIGERS, J.D.; THOMAS, E.D. & NOWINSKI, R.C. - Monoclonal antibodies to cytomegalovirus rapid identifications of clinical isolates and preliminary use in diagnosis of cytomegalovirus pneumonia. Infect. Immun., 38: 273-281, 1982.
10. HO, M. - Cytomegalovirus biology and infection currents topics in infectious diseases. In: GREENOUGH, W.B. & MERIGAN, T.C., ed. Human cytomegalovirus infections in immunocompromised patients. New York, Plenun Publishing, 1982. p.171-204.
11. HUDSON, J.B., MISRA, V. & MOSMANN, T.R. - Cytomegalovirus infectivity: analysis of the phenomenon of centrifugal enhancement of infectivity. Virology, 72: 235-243, 1976.
12. NAQUI, S.H. & BLAIR, L.L. - Detection of cytomegalovirus antigen and antibodies in the urine in small infants and children. J. med. Virol., 18: 139-147, 1986.
13. PAYA, C.V.; WOLD, D.A. & SMITH, T.F. - Detection of cytomegalovirus infections in specimens other than urine by shell vial assay and conventional tubes cell cultures. J. clin. Microbiol., 25: 755-577, 1987.
14. REYNOLDS, D.W.; STAGNO, S. & ALFORD, C.A. - Diagnostic procedures for viral, rickettsial and chlamydial infections. In: LENNETTE, E.H. & SCHMIDT N.J., ed. - Laboratory diagnosis of cytomegalovirus infections. 5th. ed. Washington, American Public Health Association, 1979. p.399-439.
15. SPECTOR, S.A.; RUA, J.A.; SPECTOR, D.H. & MCMILLIAN, R. - Detection of human cytomegalovirus in clinical specimens by DNA DNA hibridisation. J. infect. Dis., 150: 121-126, 1984.
16. SWENSON, P.D. & KAPLAN, N.H. - Rapid detection of cytomegalovirus in cell cultures by indirect immunoperoxidase stainning with monoclonal antibodies to an early nuclear antigen. J. clin. Microbiol., 21: 669-673, 1985.
17. TYMS, A.S.; TAYLOR, D.L. & PARKIN, J.M. - Cytomegalovirus and the Acquired Immunodeficiency Syndrome. J. Antimicrob. Chemother., 23: 89-105, 1989.
18. VANDERBY, W.; TORENSMA, R.; VANSON, W.J.; ANEMA, J.; SCHIRM, J.; TEGZESS, A.M. & THE, A.M. - Rapid immunodiagnosis of active cytomegalovirus infection by monoclonal antibodies staining by blood leucocytes. J. med. Virol., 25: 179-188, 1988.
19. YOW, M.D. - Congenital cytomegalovirus diseases. A new problem. J. infect. Dis., 159: 163-167, 1989.

Dirección para correspondencia:

Dra. M. Alvarez

Instituto de Medicina Tropical "Pedro Kourí"

Apartado Postal Nº 601

Marianao, C. Habana, Cuba

Fechas de Publicación

Publicación en esta colección
12 Set 2006
Fecha del número
Jun 1991

Histórico

Acepto
18 Mar 1991
Recibido
16 Mayo 1990

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

[1] 1. ALPERT, G.; MAZERON M.C.; COLIMON, R. & PLOTKIN, S. - Rapid detection of human cytomegalovirus in the urine of humans. J. infect. Dis., 152: 631-633, 1985.

[2] 2. ANDERSON, C.H. & MICHAELS, R.H. - Cytomegalovirus infections detection by direct fluorescent-antibody technique. Lancet, 2: 308-309, 1987.

[3] 3. BLASER, J.M. & COHN, D.L. - Opportunistic infections in patients with AIDS. Clues to the epidemiology of AIDS and the relative virulence of pathogens. J. infect. Dis., 8: 21-30, 1986.

[4] 4. COREY, L.; DRAVAGON, J. & BENJAMIN, D. - Rapid methods and automation in microbiology. In: TILTON, R.C., ed - Immunoperoxidase stainning in the clinical virology laboratory, Washington, American Society for Microbiology. p.246-253.

[5] 5. DEMMLER, G.J.; BUFFONE, G.J.; SCHIMBOR, C.M. & MAY, R.A. - Detection of cytomegalovirus in urine from newborns by using polymerase chain reaction DNA amplification. J. infect. Dis., 158: 1177-1184, 1988.

[6] 6. FORBES, B.A. - Acquisition of cytomegalovirus infections. An update. Clin. Microbiol. Rev., 2: 202-261, 1989.

[7] 7. GLEAVES, C.A.; SMITH, T.F.; SHUSTER, E.A. & PEARSON, G.R. - Rapid detection of cytomegalovirus in MRC 5 cells inoculated with urine specimens by using low-speed centrifugation and monoclonal antibody to an early antigen. J. clin. Microbiol., 19: 917-919, 1984.

[8] 8. GLENN, J. - Cytomegalovirus infections following renal transplantation. Rev. infect. Dis., 3: 1151-1178, 1981.

[9] 9. GOLDSTEIN, L.C.; DOUGALL, J.K.; HACKMAN, R.C.; MIGERS, J.D.; THOMAS, E.D. & NOWINSKI, R.C. - Monoclonal antibodies to cytomegalovirus rapid identifications of clinical isolates and preliminary use in diagnosis of cytomegalovirus pneumonia. Infect. Immun., 38: 273-281, 1982.

[10] 10. HO, M. - Cytomegalovirus biology and infection currents topics in infectious diseases. In: GREENOUGH, W.B. & MERIGAN, T.C., ed. Human cytomegalovirus infections in immunocompromised patients. New York, Plenun Publishing, 1982. p.171-204.

[11] 11. HUDSON, J.B., MISRA, V. & MOSMANN, T.R. - Cytomegalovirus infectivity: analysis of the phenomenon of centrifugal enhancement of infectivity. Virology, 72: 235-243, 1976.

[12] 12. NAQUI, S.H. & BLAIR, L.L. - Detection of cytomegalovirus antigen and antibodies in the urine in small infants and children. J. med. Virol., 18: 139-147, 1986.

[13] 13. PAYA, C.V.; WOLD, D.A. & SMITH, T.F. - Detection of cytomegalovirus infections in specimens other than urine by shell vial assay and conventional tubes cell cultures. J. clin. Microbiol., 25: 755-577, 1987.

[14] 14. REYNOLDS, D.W.; STAGNO, S. & ALFORD, C.A. - Diagnostic procedures for viral, rickettsial and chlamydial infections. In: LENNETTE, E.H. & SCHMIDT N.J., ed. - Laboratory diagnosis of cytomegalovirus infections. 5th. ed. Washington, American Public Health Association, 1979. p.399-439.

[15] 15. SPECTOR, S.A.; RUA, J.A.; SPECTOR, D.H. & MCMILLIAN, R. - Detection of human cytomegalovirus in clinical specimens by DNA DNA hibridisation. J. infect. Dis., 150: 121-126, 1984.

[16] 16. SWENSON, P.D. & KAPLAN, N.H. - Rapid detection of cytomegalovirus in cell cultures by indirect immunoperoxidase stainning with monoclonal antibodies to an early nuclear antigen. J. clin. Microbiol., 21: 669-673, 1985.

[17] 17. TYMS, A.S.; TAYLOR, D.L. & PARKIN, J.M. - Cytomegalovirus and the Acquired Immunodeficiency Syndrome. J. Antimicrob. Chemother., 23: 89-105, 1989.

[18] 18. VANDERBY, W.; TORENSMA, R.; VANSON, W.J.; ANEMA, J.; SCHIRM, J.; TEGZESS, A.M. & THE, A.M. - Rapid immunodiagnosis of active cytomegalovirus infection by monoclonal antibodies staining by blood leucocytes. J. med. Virol., 25: 179-188, 1988.

[19] 19. YOW, M.D. - Congenital cytomegalovirus diseases. A new problem. J. infect. Dis., 159: 163-167, 1989.

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