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Updated: Jul 4, 2026

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
Published on: November 1, 2018
An alternative method for the synthesis of competitor RNA transcripts useful for specific detection and quantitation
Gilberto Vaughan1, Yolanda Gonzalez-Hernandez, Jose-Carmen Gudino
1Department of Immunology Research, Institute for Epidemiological Diagnosis and Reference, Secretariat of Health, Carpio 470, Col. Santo Tomas, Mexico City 11340, Mexico.
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.
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