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An alternative method for the synthesis of competitor RNA transcripts useful for specific detection and quantitation of dengue virus serotype 2 genome and replicative intermediate RNA.
Vaughan, Gilberto; Gonzalez-Hernandez, Yolanda; Gudino, Jose-Carmen; Olivera, Hiram; Landa-Piedra, Abraham; Escobar-Gutierrez, Alejandro.
Afiliação
  • Vaughan G; Department of Immunology Research, Institute for Epidemiological Diagnosis and Reference, Secretariat of Health, Carpio 470, Col. Santo Tomas, Mexico City 11340, Mexico.
J Virol Methods ; 152(1-2): 72-6, 2008 Sep.
Article em En | MEDLINE | ID: mdl-18597860
The development of a quantitative-competitive reverse transcription-PCR (RT-PCR) assay to quantify dengue virus (DEN) genome (vRNA) and its replicative intermediate RNA (vRI) is described. A highly conserved region located on the DEN capsid-premembrane genes was used to produce a competitor RNA molecule which contains an internal deletion of 70 nucleotides. The competitor provides a suitable internal control useful to quantify viral RNA from all four dengue virus (DEN 1-4) serotypes. The detection limit of the assay was found to be 100 copies per reaction. This is a rapid, simple, sensitive, inexpensive and easy method for quantitation of DEN RNA species.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA Viral / Genoma Viral / Reação em Cadeia da Polimerase Via Transcriptase Reversa / Dengue / Vírus da Dengue Tipo de estudo: Diagnostic_studies Limite: Animals / Humans Idioma: En Revista: J Virol Methods Ano de publicação: 2008 Tipo de documento: Article País de afiliação: México País de publicação: Holanda

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA Viral / Genoma Viral / Reação em Cadeia da Polimerase Via Transcriptase Reversa / Dengue / Vírus da Dengue Tipo de estudo: Diagnostic_studies Limite: Animals / Humans Idioma: En Revista: J Virol Methods Ano de publicação: 2008 Tipo de documento: Article País de afiliação: México País de publicação: Holanda