Suscriptores   Asóciese  
Textos Completos Autorizados
DEFINIÇAO DOS VARIOS PARAMETROS DA REAÇAO EM CADEIA DA POLIMERASE PARA O DIAGNOSTICO DA DENGUE

v-estudie-lo-mejor.gif
Expertos Invitados





DEFINIÇAO DOS VARIOS PARAMETROS DA REAÇAO EM CADEIA DA POLIMERASE PARA O DIAGNOSTICO DA DENGUE

(especial para SIIC © Derechos reservados)
Dengue é a mais importante doença causada por um arbovírus no mundo. O desenvolvimento de novos métodos diagnósticos ou a melhoria dos métodos já disponíveis certamente resultarão em um melhor cuidado dos pacientes que apresentam esta doença
lopesda9.jpg Autor:
Lopes Da Fonseca, Benedito Ant
Columnista Experto de SIIC

Institución:
Departamento de Clínica Médica Facultad de Medicina de Ribeirão Preto Universidad de São Paulo SP, Brasil

Artículos publicados por Lopes Da Fonseca, Benedito Ant
Coautor
Sérgio Oliveira de Paula* 
D.V.M., M.Sc. Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo, Ribeirão Preto, São Paulo, Brasil*
Recepción del artículo
6 de Julio, 2004 		Aprobación
16 de Septiembre, 2004
Primera edición
15 de Noviembre, 2004 		Segunda edición, ampliada y corregida
7 de Junio, 2021

DEFINIÇAO DOS VARIOS PARAMETROS DA REAÇAO EM CADEIA DA POLIMERASE PARA O DIAGNOSTICO DA DENGUE

Resumen
Dengue é a mais importante doença causada por um arbovírus no mundo. O desenvolvimento de novos métodos diagnósticos ou a melhoria dos métodos já disponíveis certamente resultarão em um melhor cuidado dos pacientes que apresentam esta doença. Recentemente, nós temos trabalhado, em nosso laboratório, na definição de parâmetros que melhorem o diagnóstico molecular da dengue. Nesta revisão, nós discutimos a padronização das várias etapas da técnica da reação em cadeia da polimerase (PCR) visando a definição de qual é a melhor amostra clínica a ser usada no diagnóstico da dengue; qual é a melhor técnica de extração do RNA viral de amostras clínicas; e qual é o melhor kit usado regularmente nos ensaios de RT-PCR para o diagnóstico da dengue. Nós também desenhamos uma nested-PCR, baseada em um protocolo publicado anteriormente e mostramos que esta técnica pode ser usada com sucesso na detecção e diferenciação dos vírus dengue presentes em amostras clínicas que contenham IgM. Finalmente, nós melhoramos a capacidade de detecção de infecções pelos vírus dengue em células C6/36 através do uso da PCR ao invés da IFA. Resumidamente, nossos dados mostram que esta técnica pode substituir o isolamento e identificação dos vírus dengue no futuro.

Palabras clave
Dengue, PCR, diagnóstico


Artículo completo
(castellano)
Extensión:  +/-5.06 páginas impresas en papel A4
Exclusivo para suscriptores/assinantes

OPTIMIZATION OF PCR PROTOCOLS FOR DENGUE DIAGNOSIS

Abstract
Dengue is the most important arboviral disease transmitted to humans. Development of new diagnostic methods or improvement of old ones will result in a better management of this disease. Currently, in our laboratory, we have been working on the improvement of the molecular diagnosis of this disease. In this review, we discuss the standardization of several steps of the polymerase chain reaction (PCR) aiming at the definition of which is the best technique for extracting viral RNA from human samples; which is the best clinical sample to be used for dengue diagnosis; and which is the best kit regularly used on RT-PCR protocols for dengue diagnosis. We have also designed a nested RT-PCR, based on previously published protocol and showed that this technique may be successfully used on the detection and differentiation of dengue viruses present in IgM-containing samples. Finally we have improved the detection rate of dengue virus detection on C6/36 cells by using PCR instead of IFA for detection of infection. In summary, our data show that this technique could replace dengue virus isolation and identification in the future due to its ease of manipulation and the rapidity of virus identification.

Key words
Dengue, PCR, diagnostic

Clasificación en siicsalud
Artículos originales > Expertos de Iberoamérica >
página   www.siicsalud.com/des/expertocompleto.php/

Especialidades
Principal: Infectología
Relacionadas: Bioquímica, Diagnóstico por Laboratorio, Epidemiología



Comprar este artículo
Extensión: 5.06 páginas impresas en papel A4

file05.gif (1491 bytes) Artículos seleccionados para su compra



Enviar correspondencia a:
Lopes da Fonseca, Benedito Antônio
Bibliografía del artículo
  1. Anônimo. Dengue hemorragic fever, diagnosis, treatment and control. World Health Organization. Geneva, Switzerland. 1998.
  2. Morbidity and Mortality Weekly Report. Dengue type 3 infection. Nicaragua and Panamá, October/November 1994. Morbid. Mortal. Weekly Rep., 44(2):21-24. 1995.
  3. Miagostovich MP, Santos FB, Simone TS, Costa EV, Filippis AM, Schatzmayr HG, Nogueira RMR. Genetic characterization of dengue virus type 3 isolates in the state of Rio de Janeiro, 2001. Braz. J. Med. Biol. Res., 35(8), 869-872. 2002.
  4. Henchal E, Putnak JR. The dengue viruses. Clin. Microbiol. Rev., 3:376-396. 1990.
  5. Takeda A, Ennis FA. FcR-mediated enhancement of HIV-1 infection by antibody. AIDS Res. Hum. Retroviruses, 6: 999-1004. 1990.
  6. Halstead SB. Pathogenesis of dengue: challenges to molecular biology. Science, 239: 476-481. 1988.
  7. Anônimo. Pan American Health Organization. Dengue and dengue hemorragic fever in the Americas: guidelines for prevention and control. Scientific publication 548. Pan American Health Organization, Washington, D.C. 1994.
  8. Guzman MG, Kouri G. Advances in dengue diagnosis. Clin. Diagn. Lab. Immnunol., 3:621-627. 1996.
  9. Deubel V. The contribution of molecular techniques to the diagnosis of dengue infection. In DJ Gubler and G Kuno. Dengue and dengue hemmorrhagic fever. Cab International, London, United Kingdom, pp 335-366. 1997.
  10. Deubel V, Laille M, Hugnot JP, Chungue E, Guesdon JL, Drouet MT, Bassot S, Chevrier D. Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue vírus serotypes in peripheral blood. J. Virol. Methods, 30: 41-54. 1990.
  11. Henchal EA, Polo SI, Vorndam V, Yaemsiri C, Innis BI, Hoke CH. Sensitivity and specificity of a universal primer set for the rapid diagnosis of dengue virus infections by polymerase chain reaction and nucleic acid hibridization. Am. J. Med. Hyg., 45(4):418-428. 1991.
  12. Morita K, Meomoto S, Honda S, Onishi K, Murata M, Tanaka M, Igarashi A. Rapid detection of virus genome from imported dengue fever and dengue hemorrhagic fever patients by direct polymease chain reaction . J. Med. Virol., 44: 54-58. 1994.
  13. Lanciotti RS, Calisher CH, Gubler DG, Chang G, Vordam V. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J. Clin. Microbiol., 30: 545-551. 1992.
  14. Pao CC, Yao DS, Lin CY, King CC. Amplification of viral RNA for the detection of dengue types 1 and 2 virus. J. Infect., 24(1):23-29. 1992.
  15. Chippaux A, Deubel V. Application of the polymerase chain reaction (PCR) to diagnosis in virology. Bull. Soc. Pathol. Exot., 84(5):704-711. 1991.
  16. Harris E, Sandoval E, Xet-Mull AM, Johnson M, Riley LW. Rapid subtyping of dengue viruses by restriction site-specific (RSS)-PCR. Virology, 253: 86-95. 1999.
  17. Figueiredo LT, Batista WC, Igarashi A. Detection and identification of dengue virus isolates from Brazil by a simplified reverse transcription – polymerase chain reaction (RT-PCR) method. Rev. Inst. Med. Trop. São Paulo, 39(2): 79-83. 1997.
  18. Pao CC, Yao DS, Lin CY, King CC. Amplification of viral RNA for the detection of dengue types 1 and 2 virus. J. Infect., 24(1):23-29. 1992.
  19. Suk-Yin C, Kautner I, Sai-Kit L. Detection and serotyping of dengue viruses by PCR: a simple, rapid method for the isolation of viral RNA from infected mosquito larvae. South Asian J. Trop. Med. Public Health, 25: 258-261. 1994.
  20. De Paula SO, Pires Neto RJ, Corrêa JA, Assumpção SR, Costa ML, Lima DM, Fonseca BA. The use of reverse transcription-polymerase chain reaction (RT-PCR) for the rapid detection and identification of dengue virus in an endemic region: a validation study. Trans. R. S. Trop. Med. Hyg., 96:266-269. 2002.
  21. De Paula SO, Nunes C, Matos R, Machado Z, Malta DL, Fonseca BA. Comparison of techniques for extracting viral RNA from isolation-negative serum for dengue diagnosis by polymerase chain reaction. J. Virol. Methods, 98 (2): 119-125. 2001.
  22. De Paula SO, Fonseca BA. Optimizing dengue diagnosis by RT-PCR in IgM-positive samples: comparison of whole blood, buffy-coat and serum as clinical samples. J. Virol. Methods, 102: 113-117. 2002.
  23. De Paula SO, Lima DM, Clotteau M, Pires Neto RJ, Fonseca BA. Improved detection of dengue-1 virus from igm-positive serum samples using C6/36 cell cultures in association with RT-PCR. Intervirology, 46(4): 227-31. 2003.
  24. Lanciotti RS, Calisher CH, Gubler DG, Chang G, Vordam V. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J. Clin. Microbiol., 30: 545-551. 1992.
  25. Vorndam V, Kuno G, Rosado N. A PCR-restriction enzyme techinique for determining dengue vírus subgroups within serotypes. J. Virol. Methods, 48: 237-244. 1994.
  26. Vorndam V, Nogueira RM, Trent DW. Restriction enzyme analysis of American region dengue viruses. Arch. Virol., 136: 191-196. 1994.
  27. De Paula SO, Malta DL, Fonseca BA. Detection and identification of dengue-1 virus from clinical samples by a nested-PCR followed by restriction enzyme digestion of amplicons. J. Med. Virol., 66 (4): 529-534. 2002.
  28. De Paula SO, Lima CM, Torres MP, Pereira MR, Fonseca BA. One-step RT-PCR protocols improves the rate of dengue diagnosis compared with the two-step approach. J. Clin. Virol., 30: 297-301, 2004.
  29. Warrilow D, Northill JA, Pyke A, Smith Greg A. Single Rapid TaqMan fluorogenic probe based PCR assay that detects all four dengue serotypes. J. Med. Virol. 66: 524-528, 2002.
  30. Houng HS, Chen RC, Vaughan DW, Kanesa-Thasan N. Development of a fluorogenic RT-PCR systems for quantitative identification of dengue virus serotypes 1-4 using conserved and serotype-specific 3´noncoding sequences. J. Virol. Methods 95: 19-32, 2001.
 
 
 
 
 
 
 
 
 
 
 
 
Está expresamente prohibida la redistribución y la redifusión de todo o parte de los contenidos de la Sociedad Iberoamericana de Información Científica (SIIC) S.A. sin previo y expreso consentimiento de SIIC.
ua31618
Acerca de SIIC   Estructura de SIIC   Fuentes informativas  

Sociedad Iberoamericana de Información Científica

Copyright siicsalud© 1997-2024 ISSN siicsalud: 1667-9008
ua31517