Detection of Middle East respiratory syndrome coronavirus using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
Identifieur interne : 001C84 ( Main/Exploration ); précédent : 001C83; suivant : 001C85Detection of Middle East respiratory syndrome coronavirus using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
Auteurs : Kazuya Shirato [Japon] ; Takuya Yano ; Syouhei Senba [Japon] ; Shigehiro Akachi ; Takashi Kobayashi ; Takamichi Nishinaka ; Tsugunori Notomi [Japon] ; Shutoku Matsuyama [Japon]Source :
- Virology Journal [ 1743-422X ] ; 2014.
Descripteurs français
- KwdFr :
- Alignement de séquences, Analyse de séquence d'ADN, Animaux, Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification), Données de séquences moléculaires, Gènes viraux, Humains, Infections à coronavirus (diagnostic), Infections à coronavirus (virologie), Projets pilotes, Protéines nucléocapside (génétique), Reproductibilité des résultats, Réaction de polymérisation en chaine en temps réel, Sensibilité et spécificité, Séquence nucléotidique, Techniques d'amplification d'acides nucléiques.
- MESH :
- diagnostic : Infections à coronavirus.
- génétique : Coronavirus du syndrome respiratoire du Moyen-Orient, Protéines nucléocapside.
- isolement et purification : Coronavirus du syndrome respiratoire du Moyen-Orient.
- virologie : Infections à coronavirus.
- Alignement de séquences, Analyse de séquence d'ADN, Animaux, Données de séquences moléculaires, Gènes viraux, Humains, Projets pilotes, Reproductibilité des résultats, Réaction de polymérisation en chaine en temps réel, Sensibilité et spécificité, Séquence nucléotidique, Techniques d'amplification d'acides nucléiques.
English descriptors
- KwdEn :
- Animals, Base Sequence, Coronavirus Infections (diagnosis), Coronavirus Infections (virology), Genes, Viral, Humans, Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (isolation & purification), Molecular Sequence Data, Nucleic Acid Amplification Techniques, Nucleocapsid Proteins (genetics), Pilot Projects, Real-Time Polymerase Chain Reaction, Reproducibility of Results, Sensitivity and Specificity, Sequence Alignment, Sequence Analysis, DNA.
- MESH :
- chemical , genetics : Nucleocapsid Proteins.
- diagnosis : Coronavirus Infections.
- genetics : Middle East Respiratory Syndrome Coronavirus.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus.
- virology : Coronavirus Infections.
- Animals, Base Sequence, Genes, Viral, Humans, Molecular Sequence Data, Nucleic Acid Amplification Techniques, Pilot Projects, Real-Time Polymerase Chain Reaction, Reproducibility of Results, Sensitivity and Specificity, Sequence Alignment, Sequence Analysis, DNA.
Abstract
The first documented case of Middle East Respiratory Syndrome coronavirus (MERS-CoV) occurred in 2012, and outbreaks have continued ever since, mainly in Saudi Arabia. MERS-CoV is primarily diagnosed using a real-time RT-PCR assay, with at least two different genomic targets required for a positive diagnosis according to the case definition of The World Health Organization (WHO) as of 3 July 2013. Therefore, it is urgently necessary to develop as many specific genetic diagnostic methods as possible to allow stable diagnosis of MERS-CoV infections.
Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) is a genetic diagnostic method used widely for the detection of viral pathogens, which requires only a single temperature for amplification, and can be completed in less than 1 h. This study developed a novel RT-LAMP assay for detecting MERS-CoV using primer sets targeting a conserved nucleocapsid protein region.
The RT-LAMP assay was capable of detecting as few as 3.4 copies of MERS-CoV RNA, and was highly specific, with no cross-reaction to other respiratory viruses. Pilot experiments to detect MERS-CoV from medium containing pharyngeal swabs inoculated with pre-titrated viruses were also performed. The RT-LAMP assay exhibited sensitivity similar to that of MERS-CoV real-time RT-PCR.
These results suggest that the RT-LAMP assay described here is a useful tool for the diagnosis and epidemiologic surveillance of human MERS-CoV infections.
The online version of this article (doi:10.1186/1743-422X-11-139) contains supplementary material, which is available to authorized users.
Url:
DOI: 10.1186/1743-422X-11-139
PubMed: 25103205
PubMed Central: 4132226
Affiliations:
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Le document en format XML
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Alignment</term><term>Sequence Analysis, DNA</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Alignement de séquences</term><term>Analyse de séquence d'ADN</term><term>Animaux</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (génétique)</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification)</term><term>Données de séquences moléculaires</term><term>Gènes viraux</term><term>Humains</term><term>Infections à coronavirus (diagnostic)</term><term>Infections à coronavirus (virologie)</term><term>Projets pilotes</term><term>Protéines nucléocapside (génétique)</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaine en temps réel</term><term>Sensibilité et spécificité</term><term>Séquence nucléotidique</term><term>Techniques d'amplification d'acides nucléiques</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Coronavirus Infections</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Infections à coronavirus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term><term>Protéines nucléocapside</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Infections à coronavirus</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Coronavirus Infections</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Genes, Viral</term><term>Humans</term><term>Molecular Sequence Data</term><term>Nucleic Acid Amplification Techniques</term><term>Pilot Projects</term><term>Real-Time Polymerase Chain Reaction</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Sequence Alignment</term><term>Sequence Analysis, DNA</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Alignement de séquences</term><term>Analyse de séquence d'ADN</term><term>Animaux</term><term>Données de séquences moléculaires</term><term>Gènes viraux</term><term>Humains</term><term>Projets pilotes</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaine en temps réel</term><term>Sensibilité et spécificité</term><term>Séquence nucléotidique</term><term>Techniques d'amplification d'acides nucléiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>The first documented case of Middle East Respiratory Syndrome coronavirus (MERS-CoV) occurred in 2012, and outbreaks have continued ever since, mainly in Saudi Arabia. MERS-CoV is primarily diagnosed using a real-time RT-PCR assay, with at least two different genomic targets required for a positive diagnosis according to the case definition of The World Health Organization (WHO) as of 3 July 2013. Therefore, it is urgently necessary to develop as many specific genetic diagnostic methods as possible to allow stable diagnosis of MERS-CoV infections.</p></sec><sec><title>Methods</title><p>Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) is a genetic diagnostic method used widely for the detection of viral pathogens, which requires only a single temperature for amplification, and can be completed in less than 1 h. This study developed a novel RT-LAMP assay for detecting MERS-CoV using primer sets targeting a conserved nucleocapsid protein region.</p></sec><sec><title>Results</title><p>The RT-LAMP assay was capable of detecting as few as 3.4 copies of MERS-CoV RNA, and was highly specific, with no cross-reaction to other respiratory viruses. Pilot experiments to detect MERS-CoV from medium containing pharyngeal swabs inoculated with pre-titrated viruses were also performed. The RT-LAMP assay exhibited sensitivity similar to that of MERS-CoV real-time RT-PCR.</p></sec><sec><title>Conclusions</title><p>These results suggest that the RT-LAMP assay described here is a useful tool for the diagnosis and epidemiologic surveillance of human MERS-CoV infections.</p></sec><sec><title>Electronic supplementary material</title><p>The online version of this article (doi:10.1186/1743-422X-11-139) contains supplementary material, which is available to authorized users.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Danielsson, N" uniqKey="Danielsson N">N Danielsson</name></author><author><name sortKey="Catchpole, M" uniqKey="Catchpole M">M Catchpole</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zaki, Am" uniqKey="Zaki A">AM Zaki</name></author><author><name sortKey="Van Boheemen, S" uniqKey="Van Boheemen S">S van Boheemen</name></author><author><name sortKey="Bestebroer, 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Wendtner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Baric, Rs" uniqKey="Baric R">RS Baric</name></author><author><name sortKey="Stohlman, Sa" uniqKey="Stohlman S">SA Stohlman</name></author><author><name sortKey="Lai, Mm" uniqKey="Lai M">MM Lai</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lai, Mm" uniqKey="Lai M">MM Lai</name></author><author><name sortKey="Patton, Cd" uniqKey="Patton C">CD Patton</name></author><author><name sortKey="Baric, Rs" uniqKey="Baric R">RS Baric</name></author><author><name sortKey="Stohlman, Sa" uniqKey="Stohlman S">SA Stohlman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Spaan, Wj" uniqKey="Spaan W">WJ Spaan</name></author><author><name sortKey="Rottier, Pj" uniqKey="Rottier P">PJ Rottier</name></author><author><name sortKey="Horzinek, Mc" uniqKey="Horzinek M">MC Horzinek</name></author><author><name sortKey="Van Der Zeijst, Ba" uniqKey="Van Der Zeijst B">BA van der Zeijst</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Japon</li></country><region><li>Région de Kantō</li></region><settlement><li>Tokyo</li></settlement></list><tree><noCountry><name sortKey="Akachi, Shigehiro" sort="Akachi, Shigehiro" uniqKey="Akachi S" first="Shigehiro" last="Akachi">Shigehiro Akachi</name><name sortKey="Kobayashi, Takashi" sort="Kobayashi, Takashi" uniqKey="Kobayashi T" first="Takashi" last="Kobayashi">Takashi Kobayashi</name><name sortKey="Nishinaka, Takamichi" sort="Nishinaka, Takamichi" uniqKey="Nishinaka T" first="Takamichi" last="Nishinaka">Takamichi Nishinaka</name><name sortKey="Yano, Takuya" sort="Yano, Takuya" uniqKey="Yano T" first="Takuya" last="Yano">Takuya Yano</name></noCountry><country name="Japon"><region name="Région de Kantō"><name sortKey="Shirato, Kazuya" sort="Shirato, Kazuya" uniqKey="Shirato K" first="Kazuya" last="Shirato">Kazuya Shirato</name></region><name sortKey="Matsuyama, Shutoku" sort="Matsuyama, Shutoku" uniqKey="Matsuyama S" first="Shutoku" last="Matsuyama">Shutoku Matsuyama</name><name sortKey="Notomi, Tsugunori" sort="Notomi, Tsugunori" uniqKey="Notomi T" first="Tsugunori" last="Notomi">Tsugunori Notomi</name><name sortKey="Senba, Syouhei" sort="Senba, Syouhei" uniqKey="Senba S" first="Syouhei" last="Senba">Syouhei Senba</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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