title: 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.
abstract: 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.
authors.affiliation: Department of Immunology Research, Institute for Epidemiological Diagnosis and Reference, Secretariat of Health, Carpio 470, Col. Santo Tomas, Mexico City 11340, Mexico.
authors.forename: Gilberto
authors.initials: G
authors.lastname: Vaughan
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Gonzalez-Hernandez
authors.forename: Jose-Carmen
authors.initials: JC
authors.lastname: Gudino
authors.forename: Hiram
authors.initials: H
authors.lastname: Olivera
authors.forename: Abraham
authors.initials: A
authors.lastname: Landa-Piedra
authors.forename: Alejandro
authors.initials: A
authors.lastname: Escobar-Gutierrez
mesh_terms: D000818:Animals; D002460:Cell Line; D003715:Dengue / Q000175:diagnosis / Q000821:virology*; D003716:Dengue Virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016679:Genome, Viral; D006801:Humans; D012367:RNA, Viral / Q000032:analysis* / Q000097:blood / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012703:Serotyping
chemical_list: D012367:RNA, Viral