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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
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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



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Lopes da Fonseca, Benedito Antônio
Bibliografía del artículo
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