[Detection of Middle East Respiratory Syndrome Coronavirus by Reverse-transcription Loop-Mediated Isothermal Amplification].
Identifieur interne : 001507 ( Main/Exploration ); précédent : 001506; suivant : 001508[Detection of Middle East Respiratory Syndrome Coronavirus by Reverse-transcription Loop-Mediated Isothermal Amplification].
Auteurs : Guan Li ; Kai Nie ; Dan Zhang ; Xinna Li ; Yanqun Wang ; Wenjie Tan ; Xuejun MaSource :
- Bing du xue bao = Chinese journal of virology [ 1000-8721 ] ; 2015.
Descripteurs français
- KwdFr :
- Amorces ADN (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (), Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification), Humains, Infections à coronavirus (virologie), Techniques d'amplification d'acides nucléiques (), Transcription inverse.
- MESH :
- génétique : Amorces ADN, Coronavirus du syndrome respiratoire du Moyen-Orient.
- isolement et purification : Coronavirus du syndrome respiratoire du Moyen-Orient.
- virologie : Infections à coronavirus.
- Coronavirus du syndrome respiratoire du Moyen-Orient, Humains, Techniques d'amplification d'acides nucléiques, Transcription inverse.
English descriptors
- KwdEn :
- Coronavirus Infections (virology), DNA Primers (genetics), Humans, Middle East Respiratory Syndrome Coronavirus (classification), Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (isolation & purification), Nucleic Acid Amplification Techniques (methods), Reverse Transcription.
- MESH :
- chemical , genetics : DNA Primers.
- classification : Middle East Respiratory Syndrome Coronavirus.
- genetics : Middle East Respiratory Syndrome Coronavirus.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus.
- methods : Nucleic Acid Amplification Techniques.
- virology : Coronavirus Infections.
- Humans, Reverse Transcription.
Abstract
A simple, rapid and sensitive colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) was developed for rapid detection of Middle East respiratory syndrome coronavirus (MERS-CoV). The method employed six primers that recognized sequences of a nucleocapsid gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 60 min. Products were detected through a LA-320c Loopamp Turbidimeter (real-time RT-LAMP) or visual inspection of color change by pre-addition of Hydroxynaphthol Blue dye (visual RT-LAMP). Specificity of RT-LAMP was validated by detection of several human coronaviruses and common respiratory viruses. MERS-CoV real-time RT-LAMP had a linear correlation (R2) of 0.995 at 10(3)-10(6) copies. The limit of detection of real-time RT-LAMP, visual RT-LAMP and quantitative real-time PCR was 500, 1000 and 100 copies/reaction, respectively. The established RT-LAMP assay was demonstrated to be a rapid screening tool for MERS-CoV infection, and could be suitable in resource-limited clinical sites and for field studies.
PubMed: 26470533
Affiliations:
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Le document en format XML
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last="Ma">Xuejun Ma</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2015">2015</date><idno type="RBID">pubmed:26470533</idno><idno type="pmid">26470533</idno><idno type="wicri:Area/PubMed/Corpus">001437</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001437</idno><idno type="wicri:Area/PubMed/Curation">001437</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001437</idno><idno type="wicri:Area/PubMed/Checkpoint">001274</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001274</idno><idno type="wicri:Area/Ncbi/Merge">001301</idno><idno type="wicri:Area/Ncbi/Curation">001301</idno><idno type="wicri:Area/Ncbi/Checkpoint">001301</idno><idno type="wicri:doubleKey">1000-8721:2015:Li G:detection:of:middle</idno><idno type="wicri:Area/Main/Merge">001512</idno><idno type="wicri:Area/Main/Curation">001507</idno><idno 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sort="Ma, Xuejun" uniqKey="Ma X" first="Xuejun" last="Ma">Xuejun Ma</name></author></analytic><series><title level="j">Bing du xue bao = Chinese journal of virology</title><idno type="ISSN">1000-8721</idno><imprint><date when="2015" type="published">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Coronavirus Infections (virology)</term><term>DNA Primers (genetics)</term><term>Humans</term><term>Middle East Respiratory Syndrome Coronavirus (classification)</term><term>Middle East Respiratory Syndrome Coronavirus (genetics)</term><term>Middle East Respiratory Syndrome Coronavirus (isolation & purification)</term><term>Nucleic Acid Amplification Techniques (methods)</term><term>Reverse Transcription</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Amorces ADN (génétique)</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient ()</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (génétique)</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification)</term><term>Humains</term><term>Infections à coronavirus (virologie)</term><term>Techniques d'amplification d'acides nucléiques ()</term><term>Transcription inverse</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA Primers</term></keywords><keywords scheme="MESH" qualifier="classification" xml:lang="en"><term>Middle East Respiratory Syndrome 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>Amorces ADN</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient</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="methods" xml:lang="en"><term>Nucleic Acid Amplification Techniques</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>Humans</term><term>Reverse Transcription</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term><term>Humains</term><term>Techniques d'amplification d'acides nucléiques</term><term>Transcription inverse</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A simple, rapid and sensitive colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) was developed for rapid detection of Middle East respiratory syndrome coronavirus (MERS-CoV). The method employed six primers that recognized sequences of a nucleocapsid gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 60 min. Products were detected through a LA-320c Loopamp Turbidimeter (real-time RT-LAMP) or visual inspection of color change by pre-addition of Hydroxynaphthol Blue dye (visual RT-LAMP). Specificity of RT-LAMP was validated by detection of several human coronaviruses and common respiratory viruses. MERS-CoV real-time RT-LAMP had a linear correlation (R2) of 0.995 at 10(3)-10(6) copies. The limit of detection of real-time RT-LAMP, visual RT-LAMP and quantitative real-time PCR was 500, 1000 and 100 copies/reaction, respectively. The established RT-LAMP assay was demonstrated to be a rapid screening tool for MERS-CoV infection, and could be suitable in resource-limited clinical sites and for field studies.</div></front></TEI><affiliations><list></list><tree><noCountry><name sortKey="Li, Guan" sort="Li, Guan" uniqKey="Li G" first="Guan" last="Li">Guan Li</name><name sortKey="Li, Xinna" sort="Li, Xinna" uniqKey="Li X" first="Xinna" last="Li">Xinna Li</name><name sortKey="Ma, Xuejun" sort="Ma, Xuejun" uniqKey="Ma X" first="Xuejun" last="Ma">Xuejun Ma</name><name sortKey="Nie, Kai" sort="Nie, Kai" uniqKey="Nie K" first="Kai" last="Nie">Kai Nie</name><name sortKey="Tan, Wenjie" sort="Tan, Wenjie" uniqKey="Tan W" first="Wenjie" last="Tan">Wenjie Tan</name><name sortKey="Wang, Yanqun" sort="Wang, Yanqun" uniqKey="Wang Y" first="Yanqun" last="Wang">Yanqun Wang</name><name sortKey="Zhang, Dan" sort="Zhang, Dan" uniqKey="Zhang D" first="Dan" last="Zhang">Dan Zhang</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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