About: Use of chimeric influenza viruses as a novel internal control for diagnostic rRT-PCR assays

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

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Use of chimeric influenza viruses as a novel internal control for diagnostic rRT-PCR assays
Creator	Bao, Yun Jiang, Lingli Liu, Fen Wang, Hualiang Wang, Xueliang Xiao, Yanqun
Source	Medline; PMC
abstract	Real-time quantitative reverse transcriptase polymerase chain reaction (rRT-PCR) is now widely used to detect viral pathogens in various human specimens. The application of internal controls to validate the entire process of these assays is necessary to prevent false-negative results caused by unexpected inhibition or inefficient extraction. In the present study, we describe a strategy to produce a stable internal control for rRT-PCR by packaging foreign RNA into influenza virions using plasmid-based reverse genetics technology. The envelope structure of influenza virus can effectively protect RNA segments from RNase digestion, which provides an advantage for its routine use as an internal control. Utilizing this approach, we successfully generated a recombinant influenza virus (rPR8-HCV) containing the 5′ untranslated region (5′ UTR) of the hepatitis C virus (HCV) RNA genome. After inactivation and purification, the rPR8-HCV particles were demonstrated to be RNase resistant and stable at 4 °C for at least 252 days in human plasma, with no degradation even after being frozen and thawed multiple times. These results were reproducible in the COBAS TaqMan HCV test for 164 days. Moreover, the chimeric influenza virus particles could be easily produced in embryonated eggs and were noninfectious after inactivation treatment. Additionally, this strategy could also be adapted for real-time clinical applications of other RNA targets, providing a universal approach with broad clinical applications in rRT-PCR assays.
has issue date	2015-10-16(xsd:dateTime)
bibo:doi	10.1007/s00253-015-7042-y
bibo:pmid	26474983
has license	no-cc
sha1sum (hex)	1521e960269ff578998fb4abf303a20e24541268
schema:url	https://doi.org/10.1007/s00253-015-7042-y
resource representing a document's title	Use of chimeric influenza viruses as a novel internal control for diagnostic rRT-PCR assays
has PubMed Central identifier	PMC7080162
has PubMed identifier	26474983
schema:publication	Appl Microbiol Biotechnol
resource representing a document's body	covid:1521e960269ff578998fb4abf303a20e24541268#body_text
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