Novel pan-serotype control RNA for dengue virus typing through real-time reverse transcription-polymerase chain reaction.
Dengue virus
In vitro transcription
Serotyping
qRT-PCR
Journal
Journal of virological methods
ISSN: 1879-0984
Titre abrégé: J Virol Methods
Pays: Netherlands
ID NLM: 8005839
Informations de publication
Date de publication:
09 2019
09 2019
Historique:
received:
31
01
2019
revised:
05
06
2019
accepted:
09
06
2019
pubmed:
14
6
2019
medline:
4
6
2020
entrez:
14
6
2019
Statut:
ppublish
Résumé
Dengue virus (DENV) is the causative agent of one of the most important febrile illnesses worldwide. Four DENV serotypes are responsible for a broad clinical spectrum of the disease. Positive controls are costly and required for the validation of molecular test results of DENV serotyping. In this study, we describe the in silico design of the qDENV-Control plasmid with the target sequences to oligonucleotides and probes widely used for DENV serotyping, and the subsequent production of qDENV Control RNA by T7-driven run-off in vitro transcription. The qDENV Control RNA was successfully used to validate the positive and negative DENV serotyping results, allowing its incorporation in routine in-house protocols for virologic surveillance. This Control RNA allowed the absolute quantification of viral RNA copies from unknown samples as required in several fundamental studies.
Identifiants
pubmed: 31195032
pii: S0166-0934(19)30052-7
doi: 10.1016/j.jviromet.2019.113677
pii:
doi:
Substances chimiques
DNA Primers
0
DNA Probes
0
RNA, Viral
0
Types de publication
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
113677Informations de copyright
Copyright © 2019 Elsevier B.V. All rights reserved.