About: Detection of subgenomic mRNA of feline coronavirus by real-time polymerase chain reaction based on primer-probe energy transfer (P-sg-QPCR)

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About: Detection of subgenomic mRNA of feline coronavirus by real-time polymerase chain reaction based on primer-probe energy transfer (P-sg-QPCR) 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	Detection of subgenomic mRNA of feline coronavirus by real-time polymerase chain reaction based on primer-probe energy transfer (P-sg-QPCR)
Creator	Belák, Sándor Balka, Gyula Hakhverdyan, Mikhayil Blomberg, Jonas Bálint, Ádám Farsang, Attila Hornyák, Ákos Rasmussen, Thomas
source	Elsevier; Medline; PMC
abstract	Feline infectious peritonitis is one of the most severe devastating diseases of the Felidae. Upon the appearance of clinical signs, a cure for the infected animal is impossible. Therefore rapid and proper diagnosis for both the presence of the causative agent, feline coronavirus (FCoV) and the manifestation of feline infectious peritonitis is of paramount importance. In the present work, a novel real-time RT-PCR method is described which is able to detect FCoV and to determine simultaneously the quantity of the viral RNA. The new assay combines the M gene subgenomic messenger RNA (sg-mRNA) detection and the quantitation of the genome copies of FCoV. In order to detect the broadest spectrum of potential FCoV variants and to achieve the most accurate results in the detection ability the new assay is applying the primer-probe energy transfer (PriProET) principle. This technology was chosen since PriProET is very robust to tolerate the nucleotide substitutions in the target area. Therefore, this technology provides a very broad-range system, which is able to detect simultaneously many variants of the virus(es) even if the target genomic regions show large scale of variations. The detection specificity of the new assay was proven by positive amplification from a set of nine different FCoV strains and negative from the tested non-coronaviral targets. Examination of faecal samples of healthy young cats, organ samples of perished animals, which suffered from feline infectious peritonitis, and cat leukocytes from uncertain clinical cases were also subjected to the assay. The sensitivity of the P-sg-QPCR method was high, since as few as 10 genome copies of FCoV were detected. The quantitative sg-mRNA detection method revealed more than 10–50,000 times increase of the M gene sg-mRNA in organ materials of feline infectious peritonitis cases, compared to those of the enteric FCoV variants present in the faeces of normal, healthy cats. These results indicate the applicability of the new P-sg-QPCR test as a powerful novel tool for the better detection and quantitation of FCoV and for the improved diagnosis of feline infectious peritonitis, this important disease of the Felidae, causing serious losses in the cat populations at a global scale.
has issue date	2012-02-18(xsd:dateTime)
bibo:doi	10.1016/j.jviromet.2012.01.022
bibo:pmid	22349594
has license	no-cc
sha1sum (hex)	1546d1e182af6c416abe66319b6464083d730a5c
schema:url	https://doi.org/10.1016/j.jviromet.2012.01.022
resource representing a document's title	Detection of subgenomic mRNA of feline coronavirus by real-time polymerase chain reaction based on primer-probe energy transfer (P-sg-QPCR)
has PubMed Central identifier	PMC7112857
has PubMed identifier	22349594
schema:publication	J Virol Methods
resource representing a document's body	covid:1546d1e182af6c416abe66319b6464083d730a5c#body_text
is schema:about of	named entity 'impossible' named entity 'detect' named entity 'nucleotide' named entity 'peritonitis' named entity 'RNA' named entity 'spectrum' named entity 'subgenomic' named entity 'polymerase' covid:arg/1546d1e182af6c416abe66319b6464083d730a5c named entity 'detection' named entity 'real-time' named entity 'copies' named entity 'detection' named entity 'chosen' named entity 'tolerate' named entity 'regions' named entity 'test' named entity 'causing' named entity 'faeces' named entity 'accurate' named entity 'diagnosis' named entity 'genome' named entity 'cats' named entity 'severe' named entity 'combines' named entity 'detect' named entity 'principle' named entity 'suffered' named entity 'detected' named entity 'FCoV' named entity 'nucleotide substitutions' named entity 'causative agent' named entity 'quantitation' named entity 'quantitation' named entity 'real-time RT-PCR' named entity 'FCoV' named entity 'gene' named entity 'leukocytes' named entity 'Felidae' named entity 'energy transfer' named entity 'viral RNA' named entity 'assay' named entity 'feline infectious peritonitis' named entity 'primer' named entity 'feline coronavirus' named entity 'subgenomic mRNA' named entity 'target' named entity 'area' named entity 'annealing temperature' named entity 'FIPV' named entity 'faecal samples' named entity 'Sahna' named entity 'FCoV' named entity 'feline infectious peritonitis' named entity 'w/v' named entity 'essential amino acids' named entity 'virus replication' named entity 'nucleic acid' named entity 'PCR' named entity 'genotypes' named entity 'catteries' named entity 'melting point' named entity 'sequence data' named entity 'coronavirus' named entity 'fluorescent' named entity 'faecal samples' named entity 'Titanium' named entity 'organ' named entity 'subgenomic mRNA' named entity 'quantitation' named entity 'FCoV' named entity 'feline infectious peritonitis' named entity 'RNA' named entity 'genus'

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