Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus
Identifieur interne : 000600 ( Pmc/Checkpoint ); précédent : 000599; suivant : 000601Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus
Auteurs : Kazuya Shirato [Japon] ; Shohei Semba [Japon] ; Sherif A. El-Kafrawy [Arabie saoudite] ; Ahmed M. Hassan [Arabie saoudite] ; Ahmed M. Tolah [Arabie saoudite] ; Ikuyo Takayama [Japon] ; Tsutomu Kageyama [Japon] ; Tsugunori Notomi [Japon] ; Wataru Kamitani [Japon] ; Shutoku Matsuyama [Japon] ; Esam Ibraheem Azhar [Arabie saoudite]Source :
- Journal of Virological Methods [ 0166-0934 ] ; 2018.
Abstract
Fluorescent RT-LAMP assays using quenching probes for MERS-CoV were developed. Quenching probe (QProbe) can solve the problem in turbidity monitoring mechanism. Only primer-derived signal can be monitored specifically by QProbes. Two primer sets were developed to enable to confirm MERS case by RT-LAMP only. Both sets were highly specific and sensitive in comparison with real-time RT-PCR.
Url:
DOI: 10.1016/j.jviromet.2018.05.006
PubMed: 29763640
PubMed Central: 7113683
Affiliations:
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type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Kageyama, Tsutomu" sort="Kageyama, Tsutomu" uniqKey="Kageyama T" first="Tsutomu" last="Kageyama">Tsutomu Kageyama</name><affiliation wicri:level="1"><nlm:aff id="aff0015">Influenza virus Research Center, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Influenza virus Research Center, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Notomi, Tsugunori" sort="Notomi, Tsugunori" uniqKey="Notomi T" first="Tsugunori" last="Notomi">Tsugunori Notomi</name><affiliation wicri:level="1"><nlm:aff id="aff0010">Eiken Chemical Co., Ltd., 4-19-9 Taito, Taito-ku, Tokyo 110-8408, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Eiken Chemical Co., Ltd., 4-19-9 Taito, Taito-ku, Tokyo 110-8408</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Kamitani, Wataru" sort="Kamitani, Wataru" uniqKey="Kamitani W" first="Wataru" last="Kamitani">Wataru Kamitani</name><affiliation wicri:level="1"><nlm:aff id="aff0020">Laboratory of Clinical Research on Infectious Diseases, Department of Pathogen Molecular Biology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Laboratory of Clinical Research on Infectious Diseases, Department of Pathogen Molecular Biology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871</wicri:regionArea><wicri:noRegion>Osaka 565-0871</wicri:noRegion></affiliation></author><author><name sortKey="Matsuyama, Shutoku" sort="Matsuyama, Shutoku" uniqKey="Matsuyama S" first="Shutoku" last="Matsuyama">Shutoku Matsuyama</name><affiliation wicri:level="1"><nlm:aff id="aff0005">Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Azhar, Esam Ibraheem" sort="Azhar, Esam Ibraheem" uniqKey="Azhar E" first="Esam Ibraheem" last="Azhar">Esam Ibraheem Azhar</name><affiliation wicri:level="1"><nlm:aff id="aff0025">Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia</nlm:aff><country xml:lang="fr">Arabie saoudite</country><wicri:regionArea>Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589</wicri:regionArea><wicri:noRegion>Jeddah 21589</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff0030">Medical Laboratory Technology Department, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah 21589, Saudi Arabia</nlm:aff><country xml:lang="fr">Arabie saoudite</country><wicri:regionArea>Medical Laboratory Technology Department, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah 21589</wicri:regionArea><wicri:noRegion>Jeddah 21589</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno 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coronavirus</title><author><name sortKey="Shirato, Kazuya" sort="Shirato, Kazuya" uniqKey="Shirato K" first="Kazuya" last="Shirato">Kazuya Shirato</name><affiliation wicri:level="1"><nlm:aff id="aff0005">Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Semba, Shohei" sort="Semba, Shohei" uniqKey="Semba S" first="Shohei" last="Semba">Shohei Semba</name><affiliation wicri:level="1"><nlm:aff id="aff0010">Eiken Chemical Co., Ltd., 4-19-9 Taito, Taito-ku, Tokyo 110-8408, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Eiken Chemical Co., Ltd., 4-19-9 Taito, Taito-ku, Tokyo 110-8408</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="El Kafrawy, Sherif A" sort="El Kafrawy, Sherif A" uniqKey="El Kafrawy S" first="Sherif A." last="El-Kafrawy">Sherif A. El-Kafrawy</name><affiliation wicri:level="1"><nlm:aff id="aff0025">Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia</nlm:aff><country xml:lang="fr">Arabie saoudite</country><wicri:regionArea>Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589</wicri:regionArea><wicri:noRegion>Jeddah 21589</wicri:noRegion></affiliation></author><author><name sortKey="Hassan, Ahmed M" sort="Hassan, Ahmed M" uniqKey="Hassan A" first="Ahmed M." last="Hassan">Ahmed M. Hassan</name><affiliation wicri:level="1"><nlm:aff id="aff0025">Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia</nlm:aff><country xml:lang="fr">Arabie saoudite</country><wicri:regionArea>Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589</wicri:regionArea><wicri:noRegion>Jeddah 21589</wicri:noRegion></affiliation></author><author><name sortKey="Tolah, Ahmed M" sort="Tolah, Ahmed M" uniqKey="Tolah A" first="Ahmed M." last="Tolah">Ahmed M. Tolah</name><affiliation wicri:level="1"><nlm:aff id="aff0025">Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia</nlm:aff><country xml:lang="fr">Arabie saoudite</country><wicri:regionArea>Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589</wicri:regionArea><wicri:noRegion>Jeddah 21589</wicri:noRegion></affiliation></author><author><name sortKey="Takayama, Ikuyo" sort="Takayama, Ikuyo" uniqKey="Takayama I" first="Ikuyo" last="Takayama">Ikuyo Takayama</name><affiliation wicri:level="1"><nlm:aff id="aff0015">Influenza virus Research Center, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Influenza virus Research Center, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Kageyama, Tsutomu" sort="Kageyama, Tsutomu" uniqKey="Kageyama T" first="Tsutomu" last="Kageyama">Tsutomu Kageyama</name><affiliation wicri:level="1"><nlm:aff id="aff0015">Influenza virus Research Center, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Influenza virus Research Center, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Notomi, Tsugunori" sort="Notomi, Tsugunori" uniqKey="Notomi T" first="Tsugunori" last="Notomi">Tsugunori Notomi</name><affiliation wicri:level="1"><nlm:aff id="aff0010">Eiken Chemical Co., Ltd., 4-19-9 Taito, Taito-ku, Tokyo 110-8408, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Eiken Chemical Co., Ltd., 4-19-9 Taito, Taito-ku, Tokyo 110-8408</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Kamitani, Wataru" sort="Kamitani, Wataru" uniqKey="Kamitani W" first="Wataru" last="Kamitani">Wataru Kamitani</name><affiliation wicri:level="1"><nlm:aff id="aff0020">Laboratory of Clinical Research on Infectious Diseases, Department of Pathogen Molecular Biology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Laboratory of Clinical Research on Infectious Diseases, Department of Pathogen Molecular Biology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871</wicri:regionArea><wicri:noRegion>Osaka 565-0871</wicri:noRegion></affiliation></author><author><name sortKey="Matsuyama, Shutoku" sort="Matsuyama, Shutoku" uniqKey="Matsuyama S" first="Shutoku" last="Matsuyama">Shutoku Matsuyama</name><affiliation wicri:level="1"><nlm:aff id="aff0005">Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011</wicri:regionArea><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Azhar, Esam Ibraheem" sort="Azhar, Esam Ibraheem" uniqKey="Azhar E" first="Esam Ibraheem" last="Azhar">Esam Ibraheem Azhar</name><affiliation wicri:level="1"><nlm:aff id="aff0025">Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia</nlm:aff><country xml:lang="fr">Arabie saoudite</country><wicri:regionArea>Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589</wicri:regionArea><wicri:noRegion>Jeddah 21589</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff0030">Medical Laboratory Technology Department, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah 21589, Saudi Arabia</nlm:aff><country xml:lang="fr">Arabie saoudite</country><wicri:regionArea>Medical Laboratory Technology Department, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah 21589</wicri:regionArea><wicri:noRegion>Jeddah 21589</wicri:noRegion></affiliation></author></analytic><series><title level="j">Journal of Virological Methods</title><idno type="ISSN">0166-0934</idno><idno type="eISSN">1879-0984</idno><imprint><date when="2018">2018</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>Highlights</title><p><list list-type="simple" id="lis0005"><list-item id="lsti0005"><label>•</label><p id="par0005">Fluorescent RT-LAMP assays using quenching probes for MERS-CoV were developed.</p></list-item><list-item id="lsti0010"><label>•</label><p id="par0010">Quenching probe (QProbe) can solve the problem in turbidity monitoring mechanism.</p></list-item><list-item id="lsti0015"><label>•</label><p id="par0015">Only primer-derived signal can be monitored specifically by QProbes.</p></list-item><list-item id="lsti0020"><label>•</label><p id="par0020">Two primer sets were developed to enable to confirm MERS case by RT-LAMP only.</p></list-item><list-item id="lsti0025"><label>•</label><p id="par0025">Both sets were highly specific and sensitive in comparison with real-time RT-PCR.</p></list-item></list></p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Adhikari, B R" uniqKey="Adhikari B">B.R. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">J Virol Methods</journal-id><journal-id journal-id-type="iso-abbrev">J. Virol. Methods</journal-id><journal-title-group><journal-title>Journal of Virological Methods</journal-title></journal-title-group><issn pub-type="ppub">0166-0934</issn><issn pub-type="epub">1879-0984</issn><publisher><publisher-name>The Authors. Published by Elsevier B.V.</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">29763640</article-id><article-id pub-id-type="pmc">7113683</article-id><article-id pub-id-type="publisher-id">S0166-0934(18)30105-8</article-id><article-id pub-id-type="doi">10.1016/j.jviromet.2018.05.006</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus</article-title></title-group><contrib-group><contrib contrib-type="author" id="aut0005"><name><surname>Shirato</surname><given-names>Kazuya</given-names></name><email>shirato@nih.go.jp</email><xref rid="aff0005" ref-type="aff">a</xref><xref rid="cor0005" ref-type="corresp">*</xref></contrib><contrib contrib-type="author" id="aut0010"><name><surname>Semba</surname><given-names>Shohei</given-names></name><xref rid="aff0010" ref-type="aff">b</xref></contrib><contrib contrib-type="author" id="aut0015"><name><surname>El-Kafrawy</surname><given-names>Sherif A.</given-names></name><xref rid="aff0025" ref-type="aff">e</xref></contrib><contrib contrib-type="author" id="aut0020"><name><surname>Hassan</surname><given-names>Ahmed M.</given-names></name><xref rid="aff0025" ref-type="aff">e</xref></contrib><contrib contrib-type="author" id="aut0025"><name><surname>Tolah</surname><given-names>Ahmed M.</given-names></name><xref rid="aff0025" ref-type="aff">e</xref></contrib><contrib contrib-type="author" id="aut0030"><name><surname>Takayama</surname><given-names>Ikuyo</given-names></name><xref rid="aff0015" ref-type="aff">c</xref></contrib><contrib contrib-type="author" id="aut0035"><name><surname>Kageyama</surname><given-names>Tsutomu</given-names></name><xref rid="aff0015" ref-type="aff">c</xref></contrib><contrib contrib-type="author" id="aut0040"><name><surname>Notomi</surname><given-names>Tsugunori</given-names></name><xref rid="aff0010" ref-type="aff">b</xref></contrib><contrib contrib-type="author" id="aut0045"><name><surname>Kamitani</surname><given-names>Wataru</given-names></name><xref rid="aff0020" ref-type="aff">d</xref></contrib><contrib contrib-type="author" id="aut0050"><name><surname>Matsuyama</surname><given-names>Shutoku</given-names></name><xref rid="aff0005" ref-type="aff">a</xref></contrib><contrib contrib-type="author" id="aut0055"><name><surname>Azhar</surname><given-names>Esam Ibraheem</given-names></name><xref rid="aff0025" ref-type="aff">e</xref><xref rid="aff0030" ref-type="aff">f</xref></contrib></contrib-group><aff id="aff0005"><label>a</label>Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan</aff><aff id="aff0010"><label>b</label>Eiken Chemical Co., Ltd., 4-19-9 Taito, Taito-ku, Tokyo 110-8408, Japan</aff><aff id="aff0015"><label>c</label>Influenza virus Research Center, National Institute of Infectious Disease, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan</aff><aff id="aff0020"><label>d</label>Laboratory of Clinical Research on Infectious Diseases, Department of Pathogen Molecular Biology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan</aff><aff id="aff0025"><label>e</label>Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah 21589, Saudi Arabia</aff><aff id="aff0030"><label>f</label>Medical Laboratory Technology Department, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah 21589, Saudi Arabia</aff><author-notes><corresp id="cor0005"><label>⁎</label>Corresponding author at: Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National, Institute of Infectious Diseases, Murayama Branch, 4-7-1 Gakuen, Musashimurayama, Tokyo, 208-0011, Japan. <email>shirato@nih.go.jp</email></corresp></author-notes><pub-date pub-type="pmc-release"><day>12</day><month>5</month><year>2018</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><month>8</month><year>2018</year></pub-date><pub-date pub-type="epub"><day>12</day><month>5</month><year>2018</year></pub-date><volume>258</volume><fpage>41</fpage><lpage>48</lpage><history><date date-type="received"><day>23</day><month>2</month><year>2018</year></date><date date-type="rev-recd"><day>9</day><month>4</month><year>2018</year></date><date date-type="accepted"><day>10</day><month>5</month><year>2018</year></date></history><permissions><copyright-statement>© 2018 The Authors</copyright-statement><copyright-year>2018</copyright-year><license><license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p></license></permissions><abstract abstract-type="author-highlights" id="abs0005"><title>Highlights</title><p><list list-type="simple" id="lis0005"><list-item id="lsti0005"><label>•</label><p id="par0005">Fluorescent RT-LAMP assays using quenching probes for MERS-CoV were developed.</p></list-item><list-item id="lsti0010"><label>•</label><p id="par0010">Quenching probe (QProbe) can solve the problem in turbidity monitoring mechanism.</p></list-item><list-item id="lsti0015"><label>•</label><p id="par0015">Only primer-derived signal can be monitored specifically by QProbes.</p></list-item><list-item id="lsti0020"><label>•</label><p id="par0020">Two primer sets were developed to enable to confirm MERS case by RT-LAMP only.</p></list-item><list-item id="lsti0025"><label>•</label><p id="par0025">Both sets were highly specific and sensitive in comparison with real-time RT-PCR.</p></list-item></list></p></abstract><abstract id="abs0010"><p>Clinical detection of Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV) in patients is achieved using genetic diagnostic methods, such as real-time RT-PCR assay. Previously, we developed a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the detection of MERS-CoV [Virol J. 2014. 11:139]. Generally, amplification of RT-LAMP is monitored by the turbidity induced by precipitation of magnesium pyrophosphate with newly synthesized DNA. However, this mechanism cannot completely exclude the possibility of unexpected reactions. Therefore, in this study, fluorescent RT-LAMP assays using quenching probes (QProbes) were developed specifically to monitor only primer-derived signals. Two primer sets (targeting nucleocapsid and ORF1a sequences) were constructed to confirm MERS cases by RT-LAMP assay only. Our data indicate that both primer sets were capable of detecting MERS-CoV RNA to the same level as existing genetic diagnostic methods, and that both were highly specific with no cross-reactivity observed with other respiratory viruses. These primer sets were highly efficient in amplifying target sequences derived from different MERS-CoV strains, including camel MERS-CoV. In addition, the detection efficacy of QProbe RT-LAMP was comparable to that of real-time RT-PCR assay using clinical specimens from patients in Saudi Arabia. Altogether, these results indicate that QProbe RT-LAMP assays described here can be used as powerful diagnostic tools for rapid detection and surveillance of MERS-CoV infections.</p></abstract><kwd-group id="kwd0005"><title>Abbreviations</title><kwd>ADV, adenovirus</kwd><kwd>ATCC, American Type Culture Collection</kwd><kwd>BIP, backward inner primer</kwd><kwd>CoV, coronavirus</kwd><kwd>FFU, focus forming unit</kwd><kwd>FIP, forward inner primer</kwd><kwd>HBoV, human bocavirus</kwd><kwd>HCoV, human coronavirus</kwd><kwd>MERS, Middle East respiratory syndrome</kwd><kwd>MPV, metapneumovirus</kwd><kwd>N, nucleocapsid</kwd><kwd>ORF, open reading frame</kwd><kwd>PBS, phosphate-buffered saline</kwd><kwd>PIV, parainfluenza virus</kwd><kwd>PFU, plaque forming unit</kwd><kwd>QProbe or QP, quenching probe</kwd><kwd>RSV, respiratory syncytial virus</kwd><kwd>RT-LAMP, reverse transcription-loop-mediated isothermal amplification</kwd><kwd>TCID50, 50% tissue culture infectious dose</kwd><kwd>upE, upstream E</kwd></kwd-group><kwd-group id="kwd0010"><title>Keywords</title><kwd>Middle East respiratory syndrome</kwd><kwd>MERS coronavirus</kwd><kwd>Quenching probe</kwd><kwd>RT-LAMP</kwd></kwd-group></article-meta></front></pmc><affiliations><list><country><li>Arabie saoudite</li><li>Japon</li></country><region><li>Région de Kantō</li></region><settlement><li>Tokyo</li></settlement></list><tree><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="Kageyama, Tsutomu" sort="Kageyama, Tsutomu" uniqKey="Kageyama T" first="Tsutomu" last="Kageyama">Tsutomu Kageyama</name><name sortKey="Kamitani, Wataru" sort="Kamitani, Wataru" uniqKey="Kamitani W" first="Wataru" last="Kamitani">Wataru Kamitani</name><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="Semba, Shohei" sort="Semba, Shohei" uniqKey="Semba S" first="Shohei" last="Semba">Shohei Semba</name><name sortKey="Takayama, Ikuyo" sort="Takayama, Ikuyo" uniqKey="Takayama I" first="Ikuyo" last="Takayama">Ikuyo Takayama</name></country><country name="Arabie saoudite"><noRegion><name sortKey="El Kafrawy, Sherif A" sort="El Kafrawy, Sherif A" uniqKey="El Kafrawy S" first="Sherif A." last="El-Kafrawy">Sherif A. El-Kafrawy</name></noRegion><name sortKey="Azhar, Esam Ibraheem" sort="Azhar, Esam Ibraheem" uniqKey="Azhar E" first="Esam Ibraheem" last="Azhar">Esam Ibraheem Azhar</name><name sortKey="Azhar, Esam Ibraheem" sort="Azhar, Esam Ibraheem" uniqKey="Azhar E" first="Esam Ibraheem" last="Azhar">Esam Ibraheem Azhar</name><name sortKey="Hassan, Ahmed M" sort="Hassan, Ahmed M" uniqKey="Hassan A" first="Ahmed M." last="Hassan">Ahmed M. Hassan</name><name sortKey="Tolah, Ahmed M" sort="Tolah, Ahmed M" uniqKey="Tolah A" first="Ahmed M." last="Tolah">Ahmed M. Tolah</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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