Real-Time Sequence-Validated Loop-Mediated Isothermal Amplification Assays for Detection of Middle East Respiratory Syndrome Coronavirus (MERS-CoV)
Identifieur interne : 001600 ( Main/Exploration ); précédent : 001599; suivant : 001601Real-Time Sequence-Validated Loop-Mediated Isothermal Amplification Assays for Detection of Middle East Respiratory Syndrome Coronavirus (MERS-CoV)
Auteurs : Sanchita Bhadra [États-Unis] ; Yu Sherry Jiang [États-Unis] ; Mia R. Kumar [États-Unis] ; Reed F. Johnson [États-Unis] ; Lisa E. Hensley [États-Unis] ; Andrew D. Ellington [États-Unis]Source :
- PLoS ONE [ 1932-6203 ] ; 2015.
Descripteurs français
- KwdFr :
- Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification), Coronavirus du syndrome respiratoire du Moyen-Orient (pathogénicité), Génome viral, Humains, Infections à coronavirus (diagnostic), Infections à coronavirus (génétique), Infections à coronavirus (virologie), Protéines virales (génétique), Réaction de polymérisation en chaine en temps réel ().
- MESH :
- diagnostic : Infections à coronavirus.
- génétique : Coronavirus du syndrome respiratoire du Moyen-Orient, Infections à coronavirus, Protéines virales.
- isolement et purification : Coronavirus du syndrome respiratoire du Moyen-Orient.
- pathogénicité : Coronavirus du syndrome respiratoire du Moyen-Orient.
- virologie : Infections à coronavirus.
- Génome viral, Humains, Réaction de polymérisation en chaine en temps réel.
English descriptors
- KwdEn :
- Coronavirus Infections (diagnosis), Coronavirus Infections (genetics), Coronavirus Infections (virology), Genome, Viral, Humans, Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (isolation & purification), Middle East Respiratory Syndrome Coronavirus (pathogenicity), Real-Time Polymerase Chain Reaction (methods), Viral Proteins (genetics).
- MESH :
- chemical , genetics : Viral Proteins.
- diagnosis : Coronavirus Infections.
- genetics : Coronavirus Infections, Middle East Respiratory Syndrome Coronavirus.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus.
- methods : Real-Time Polymerase Chain Reaction.
- pathogenicity : Middle East Respiratory Syndrome Coronavirus.
- virology : Coronavirus Infections.
- Genome, Viral, Humans.
Abstract
The Middle East respiratory syndrome coronavirus (MERS-CoV), an emerging human coronavirus, causes severe acute respiratory illness with a 35% mortality rate. In light of the recent surge in reported infections we have developed asymmetric five-primer reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for detection of MERS-CoV. Isothermal amplification assays will facilitate the development of portable point-of-care diagnostics that are crucial for management of emerging infections. The RT-LAMP assays are designed to amplify MERS-CoV genomic loci located within the open reading frame (ORF)1a and ORF1b genes and upstream of the E gene. Additionally we applied one-step strand displacement probes (OSD) for real-time sequence-specific verification of LAMP amplicons. Asymmetric amplification effected by incorporating a single loop primer in each assay accelerated the time-to-result of the OSD-RT-LAMP assays. The resulting assays could detect 0.02 to 0.2 plaque forming units (PFU) (5 to 50 PFU/ml) of MERS-CoV in infected cell culture supernatants within 30 to 50 min and did not cross-react with common human respiratory pathogens.
Url:
DOI: 10.1371/journal.pone.0123126
PubMed: 25856093
PubMed Central: 4391951
Affiliations:
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Le document en format XML
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In light of the recent surge in reported infections we have developed asymmetric five-primer reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for detection of MERS-CoV. Isothermal amplification assays will facilitate the development of portable point-of-care diagnostics that are crucial for management of emerging infections. The RT-LAMP assays are designed to amplify MERS-CoV genomic loci located within the open reading frame (ORF)1a and ORF1b genes and upstream of the E gene. Additionally we applied one-step strand displacement probes (OSD) for real-time sequence-specific verification of LAMP amplicons. Asymmetric amplification effected by incorporating a single loop primer in each assay accelerated the time-to-result of the OSD-RT-LAMP assays. The resulting assays could detect 0.02 to 0.2 plaque forming units (PFU) (5 to 50 PFU/ml) of MERS-CoV in infected cell culture supernatants within 30 to 50 min and did not cross-react with common human respiratory pathogens.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Cotten, M" uniqKey="Cotten M">M Cotten</name></author><author><name sortKey="Lam, Tt" uniqKey="Lam T">TT Lam</name></author><author><name sortKey="Watson, Sj" uniqKey="Watson S">SJ Watson</name></author><author><name sortKey="Palser, Al" uniqKey="Palser A">AL Palser</name></author><author><name sortKey="Petrova, V" uniqKey="Petrova V">V Petrova</name></author><author><name sortKey="Grant, P" uniqKey="Grant P">P Grant</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Corman, Vm" uniqKey="Corman V">VM Corman</name></author><author><name sortKey="Eckerle, I" uniqKey="Eckerle I">I Eckerle</name></author><author><name 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W" uniqKey="Miller W">W Miller</name></author><author><name sortKey="Myers, Ew" uniqKey="Myers E">EW Myers</name></author><author><name sortKey="Lipman, Dj" uniqKey="Lipman D">DJ Lipman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Altschul, Sf" uniqKey="Altschul S">SF Altschul</name></author><author><name sortKey="Madden, Tl" uniqKey="Madden T">TL Madden</name></author><author><name sortKey="Schaffer, Aa" uniqKey="Schaffer A">AA Schaffer</name></author><author><name sortKey="Zhang, J" uniqKey="Zhang J">J Zhang</name></author><author><name sortKey="Zhang, Z" uniqKey="Zhang Z">Z Zhang</name></author><author><name sortKey="Miller, W" uniqKey="Miller W">W Miller</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Maryland</li><li>Texas</li></region><settlement><li>Austin (Texas)</li></settlement><orgName><li>Université du Texas à Austin</li></orgName></list><tree><country name="États-Unis"><region name="Texas"><name sortKey="Bhadra, Sanchita" sort="Bhadra, Sanchita" uniqKey="Bhadra S" first="Sanchita" last="Bhadra">Sanchita Bhadra</name></region><name sortKey="Ellington, Andrew D" sort="Ellington, Andrew D" uniqKey="Ellington A" first="Andrew D." last="Ellington">Andrew D. Ellington</name><name sortKey="Hensley, Lisa E" sort="Hensley, Lisa E" uniqKey="Hensley L" first="Lisa E." last="Hensley">Lisa E. Hensley</name><name sortKey="Jiang, Yu Sherry" sort="Jiang, Yu Sherry" uniqKey="Jiang Y" first="Yu Sherry" last="Jiang">Yu Sherry Jiang</name><name sortKey="Johnson, Reed F" sort="Johnson, Reed F" uniqKey="Johnson R" first="Reed F." last="Johnson">Reed F. Johnson</name><name sortKey="Kumar, Mia R" sort="Kumar, Mia R" uniqKey="Kumar M" first="Mia R." last="Kumar">Mia R. Kumar</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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