About: Development of a duplex real-time RT-qPCR assay to monitor genome replication, gene expression and gene insert stability during in vivo replication of a prototype live attenuated canine distemper virus vector encoding SIV gag

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About: Development of a duplex real-time RT-qPCR assay to monitor genome replication, gene expression and gene insert stability during in vivo replication of a prototype live attenuated canine distemper virus vector encoding SIV gag Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	Development of a duplex real-time RT-qPCR assay to monitor genome replication, gene expression and gene insert stability during in vivo replication of a prototype live attenuated canine distemper virus vector encoding SIV gag
Creator	Zhang, Xinsheng Martinez, Jennifer Arendt, Heather Sharma, Palka Coleman, John »more»
source	Elsevier; Medline; PMC
abstract	Abstract Advancement of new vaccines based on live viral vectors requires sensitive assays to analyze in vivo replication, gene expression and genetic stability. In this study, attenuated canine distemper virus (CDV) was used as a vaccine delivery vector and duplex 2-step quantitative real-time RT-PCR (RT-qPCR) assays specific for genomic RNA (gRNA) or mRNA have been developed that concurrently quantify coding sequences for the CDV nucleocapsid protein (N) and a foreign vaccine antigen (SIV Gag). These amplicons, which had detection limits of about 10 copies per PCR reaction, were used to show that abdominal cavity lymphoid tissues were a primary site of CDV vector replication in infected ferrets, and importantly, CDV gRNA or mRNA was undetectable in brain tissue. In addition, the gRNA duplex assay was adapted for monitoring foreign gene insert genetic stability during in vivo replication by analyzing the ratio of CDV N and SIV gag genomic RNA copies over the course of vector infection. This measurement was found to be a sensitive probe for assessing the in vivo genetic stability of the foreign gene insert.
has issue date	2015-03-01(xsd:dateTime)
bibo:doi	10.1016/j.jviromet.2014.11.015
bibo:pmid	25486083
has license	els-covid
sha1sum (hex)	bb8e1ed1f26d0f912f83eb789a0fc2d6dcca541f
schema:url	https://doi.org/10.1016/j.jviromet.2014.11.015
resource representing a document's title	Development of a duplex real-time RT-qPCR assay to monitor genome replication, gene expression and gene insert stability during in vivo replication of a prototype live attenuated canine distemper virus vector encoding SIV gag
has PubMed Central identifier	PMC7111484
has PubMed identifier	25486083
schema:publication	Journal of Virological Methods
resource representing a document's body	covid:bb8e1ed1f26d0f912f83eb789a0fc2d6dcca541f#body_text
is schema:about of	named entity 'PCR' named entity 'genetic' named entity 'vector' named entity 'undetectable' named entity 'RT-qPCR' »more»

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