About: Porcine epidemic diarrhea virus: Viral RNA detection and quantification using a validated one-step real time RT-PCR

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About: Porcine epidemic diarrhea virus: Viral RNA detection and quantification using a validated one-step real time RT-PCR Goto Sponge NotDistinct Permalink

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

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Porcine epidemic diarrhea virus: Viral RNA detection and quantification using a validated one-step real time RT-PCR
Creator	Blanchard, Yannick Grasland, Béatrice Brown, Paul Bigault, Lionel Bernard, Cécilia
Source	Elsevier; Medline; PMC
abstract	Since 2014, porcine epidemic diarrhea virus (PEDV) has reemerged in Europe. RT-PCR methods have been described for the detection of PEDV, but none have been validated according to a norm. In this study we described the development and validation of a SYBR™ Green one-step RT-qPCR according to the French norm NF U47-600, for the detection and quantification of PEDV viral RNA. The method was validated from sample preparation (feces or jejunum) through to nucleic acid extraction and RT-qPCR detection. Specificity and sensitivity, limit of detection (LoD), limit of quantification (LQ), linearity, intra and inter assay variability were evaluated using transcribed RNA and fecal and jejunum matrices spiked with virus. The analytical and diagnostic specificities and sensitivities of this RT-qPCR were 100% in this study. A LoD of 50 genome copies/5 µl of extract from fecal matrices spiked with virus or RNA transcript and 100 genome copies/5 µl of extract from jejunum matrices spiked with virus were obtained. The Lower LQ (LLQ) was 100 genome copies/5 µl and the Upper LQ (ULQ) 10(8) copies/5 µl. This method is the first, validated according a norm for PEDV and may serve as a global reference method to harmonize detection and quantification of PEDV viral RNA in both field and experimental settings.
has issue date	2020-05-31(xsd:dateTime)
bibo:doi	10.1016/j.jviromet.2020.113906
bibo:pmid	32485176
has license	no-cc
sha1sum (hex)	433b9f21f138515d9072fbd7d3d2789bed5a0ef8
schema:url	https://doi.org/10.1016/j.jviromet.2020.113906
resource representing a document's title	Porcine epidemic diarrhea virus: Viral RNA detection and quantification using a validated one-step real time RT-PCR
has PubMed Central identifier	PMC7261358
has PubMed identifier	32485176
schema:publication	J Virol Methods
resource representing a document's body	covid:433b9f21f138515d9072fbd7d3d2789bed5a0ef8#body_text
is schema:about of	named entity 'quantification' named entity 'reduce' named entity 'RT-PCR' named entity 'reproducibility' named entity 'Primer' named entity 'Porcine Epidemic Diarrhea Virus' named entity 'RT-PCR' named entity 'RNA' named entity 'RNA' named entity 'China' named entity 'ORFs' named entity 'sensitivity and specificity' named entity 'PEDV' named entity 'PEDV' named entity 'PEDV' named entity 'RNA' named entity 'linear regression' named entity 'melting curve analysis' named entity 'PEDV' named entity '100 %' named entity 'transcription' named entity 'PEDV' named entity 'base pairing' named entity 'TCoV' named entity 'hybridization' named entity 'agarose gel' named entity 'Diarrhea' named entity 'experimental settings' named entity 'PEDV' named entity 'vacuum' named entity 'PEDV' named entity 'ThermoFisher Scientific' named entity 'RNA' named entity 'swine influenza' named entity 'genotypes' named entity 'infectious bronchitis virus' named entity 'TCID50' named entity 'virus' named entity 'serially diluted' named entity 'Streptomycin' named entity 'animal health' named entity 'jejunum' named entity 'RNA' named entity 'genome' named entity 'limit of quantification' named entity 'porcine epidemic diarrhea virus' named entity 'animal health' named entity 'RNA transcripts' named entity 'PEDV' named entity 'jejunum' named entity 'Invitrogen' named entity 'vaccines' named entity 'Enteric virus' named entity 'transcription' named entity 'PEDV' named entity 'RNA extraction' named entity 'reverse transcription' named entity 'culture medium' named entity 'statistical bias' named entity 'RNA transcript' named entity 'RNA transcript' named entity 'jejunum' named entity 'feces' named entity 'genome' named entity 'Pedacovirus' named entity 'PEDV' named entity 'single stranded' named entity 'RNA genome' named entity 'EMEM' named entity 'Genbank' named entity 'viruses' named entity 'MgSO4' named entity 'nucleotides' named entity 'Trizol' named entity 'viral titer' named entity 'morbidity' named entity 'Nucleotide'

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