Quantification of the Middle East Respiratory Syndrome-Coronavirus RNA in Tissues by Quantitative Real-Time RT-PCR
Identifieur interne : 000202 ( Pmc/Checkpoint ); précédent : 000201; suivant : 000203Quantification of the Middle East Respiratory Syndrome-Coronavirus RNA in Tissues by Quantitative Real-Time RT-PCR
Auteurs :Source :
- MERS Coronavirus ; 2019.
Abstract
Since the emergence of the Middle East respiratory syndrome-coronavirus (MERS-CoV) in 2012, more than 2280 confirmed human infections and 800 associated deaths had been reported to the World Health Organization. MERS-CoV is a single-stranded RNA virus that belongs to the
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DOI: 10.1007/978-1-0716-0211-9_8
PubMed: 31883090
PubMed Central: 7122982
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Quantification of the Middle East Respiratory Syndrome-Coronavirus RNA in Tissues by Quantitative Real-Time RT-PCR</title></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">31883090</idno><idno type="pmc">7122982</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7122982</idno><idno type="RBID">PMC:7122982</idno><idno type="doi">10.1007/978-1-0716-0211-9_8</idno><date when="2019">2019</date><idno type="wicri:Area/Pmc/Corpus">000182</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000182</idno><idno type="wicri:Area/Pmc/Curation">000182</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000182</idno><idno type="wicri:Area/Pmc/Checkpoint">000202</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000202</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Quantification of the Middle East Respiratory Syndrome-Coronavirus RNA in Tissues by Quantitative Real-Time RT-PCR</title></analytic><series><title level="j">MERS Coronavirus</title><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p id="Par1">Since the emergence of the Middle East respiratory syndrome-coronavirus (MERS-CoV) in 2012, more than 2280 confirmed human infections and 800 associated deaths had been reported to the World Health Organization. MERS-CoV is a single-stranded RNA virus that belongs to the <italic>Coronaviridae</italic> family. MERS-CoV infection leads to a variety of clinical outcomes in humans ranging from asymptomatic and mild infection to severe acute lung injury and multi-organ failure and death. To study the pathogenesis of MERS-CoV infection and development of medical countermeasures (MCMs) for MERS, a number of genetically modified mouse models have been developed, including various versions of transgenic mice expressing the human DPP4 viral receptor. Tracking and quantifying viral infection, among others, in permissive hosts is a key endpoint for studying MERS pathogenesis and evaluating the efficacy of selected MCMs developed for MERS. In addition to quantifying infectious progeny virus which requires high-containment biosafety level (BSL)-3 laboratory, here we outlined an established real-time quantitative RT-PCR (RT-qPCR)-based procedure to unequivocally quantify MERS-CoV-specific RNAs within the lungs of infected human DPP4 (hDPP4, transgenic (hDPP4 Tg) mice under a standard BSL-2 laboratory.</p></div></front><back><div1 type="bibliography"><listBibl><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, Tm" uniqKey="Bestebroer T">TM Bestebroer</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chan, Jfw" uniqKey="Chan J">JFW Chan</name></author><author><name sortKey="Lau, Skp" uniqKey="Lau S">SKP Lau</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fehr, Ar" uniqKey="Fehr A">AR Fehr</name></author><author><name sortKey="Perlman, S" uniqKey="Perlman S">S Perlman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="De Wit, E" uniqKey="De Wit E">E de Wit</name></author><author><name sortKey="Van Doremalen, N" uniqKey="Van Doremalen N">N van Doremalen</name></author><author><name sortKey="Falzarano, D" uniqKey="Falzarano D">D Falzarano</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Coleman, Cm" uniqKey="Coleman C">CM Coleman</name></author><author><name sortKey="Matthews, Kl" uniqKey="Matthews K">KL Matthews</name></author><author><name sortKey="Goicochea, L" uniqKey="Goicochea L">L Goicochea</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cockrell, As" uniqKey="Cockrell A">AS Cockrell</name></author><author><name sortKey="Peck, Km" uniqKey="Peck K">KM Peck</name></author><author><name sortKey="Yount, Bl" uniqKey="Yount B">BL Yount</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Agrawal, As" uniqKey="Agrawal A">AS Agrawal</name></author><author><name sortKey="Garron, T" uniqKey="Garron T">T Garron</name></author><author><name sortKey="Tao, X" uniqKey="Tao X">X Tao</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tao, X" uniqKey="Tao X">X Tao</name></author><author><name sortKey="Garron, T" uniqKey="Garron T">T Garron</name></author><author><name sortKey="Agrawal, As" uniqKey="Agrawal A">AS Agrawal</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="chapter-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="publisher-id">978-1-0716-0211-9</journal-id><journal-id journal-id-type="doi">10.1007/978-1-0716-0211-9</journal-id><journal-id journal-id-type="nlm-ta">MERS Coronavirus</journal-id><journal-title-group><journal-title>MERS Coronavirus</journal-title><journal-subtitle>Methods and Protocols </journal-subtitle></journal-title-group><isbn publication-format="print">978-1-0716-0210-2</isbn><isbn publication-format="electronic">978-1-0716-0211-9</isbn></journal-meta><article-meta><article-id pub-id-type="pmid">31883090</article-id><article-id pub-id-type="pmc">7122982</article-id><article-id pub-id-type="publisher-id">8</article-id><article-id pub-id-type="doi">10.1007/978-1-0716-0211-9_8</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Quantification of the Middle East Respiratory Syndrome-Coronavirus RNA in Tissues by Quantitative Real-Time RT-PCR</article-title></title-group><contrib-group content-type="book editors"><contrib contrib-type="editor"><name><surname>Vijay</surname><given-names>Rahul</given-names></name><address><email>rahul-vijay@uiowa.edu</email></address><xref ref-type="aff" rid="Aff2"></xref></contrib><aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.214572.7</institution-id><institution-id institution-id-type="ISNI">0000 0004 1936 8294</institution-id><institution>Department of Microbiology and Immunology,</institution><institution>University of Iowa,</institution></institution-wrap>Iowa City, IA USA</aff></contrib-group><contrib-group><contrib contrib-type="author"><name><surname>Algaissi</surname><given-names>Abdullah</given-names></name><xref ref-type="aff" rid="Aff3">3</xref><xref ref-type="aff" rid="Aff4">4</xref></contrib><contrib contrib-type="author"><name><surname>Agrawal</surname><given-names>Anurodh S.</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Hashem</surname><given-names>Anwar M.</given-names></name><address><email>amhashem@kau.edu.sa</email></address><xref ref-type="aff" rid="Aff5">5</xref><xref ref-type="aff" rid="Aff6">6</xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Tseng</surname><given-names>Chien-Te K.</given-names></name><address><email>sktseng@utmb.edu</email></address><xref ref-type="aff" rid="Aff3">3</xref><xref ref-type="aff" rid="Aff8">8</xref></contrib><aff id="Aff3"><label>3</label><institution-wrap><institution-id institution-id-type="GRID">grid.176731.5</institution-id><institution-id institution-id-type="ISNI">0000 0001 1547 9964</institution-id><institution>Departments of Microbiology and Immunology,</institution><institution>University of Texas Medical Branch,</institution></institution-wrap>Galveston, TX USA</aff><aff id="Aff4"><label>4</label><institution-wrap><institution-id institution-id-type="GRID">grid.411831.e</institution-id><institution-id institution-id-type="ISNI">0000 0004 0398 1027</institution-id><institution>Department of Medical Laboratories Technology, College of Applied Medical Sciences,</institution><institution>Jazan University,</institution></institution-wrap>Jazan, Saudi Arabia</aff><aff id="Aff5"><label>5</label><institution-wrap><institution-id institution-id-type="GRID">grid.412125.1</institution-id><institution-id institution-id-type="ISNI">0000 0001 0619 1117</institution-id><institution>Department of Medical Microbiology and Parasitology, Faculty of Medicine,</institution><institution>King Abdulaziz University,</institution></institution-wrap>Jeddah, Saudi Arabia</aff><aff id="Aff6"><label>6</label><institution-wrap><institution-id institution-id-type="GRID">grid.412125.1</institution-id><institution-id institution-id-type="ISNI">0000 0001 0619 1117</institution-id><institution>Vaccines and Immunotherapy Unit, King Fahd Medical Research Center,</institution><institution>King Abdulaziz University,</institution></institution-wrap>Jeddah, Saudi Arabia</aff><aff id="Aff7"><label>7</label><institution-wrap><institution-id institution-id-type="GRID">grid.412125.1</institution-id><institution-id institution-id-type="ISNI">0000 0001 0619 1117</institution-id><institution>Special Infectious Agents Unit, King Fahd Medical Research Center,</institution><institution>King Abdulaziz University,</institution></institution-wrap>Jeddah, Saudi Arabia</aff><aff id="Aff8"><label>8</label><institution-wrap><institution-id institution-id-type="GRID">grid.176731.5</institution-id><institution-id institution-id-type="ISNI">0000 0001 1547 9964</institution-id><institution>Center for Biodefense and Emerging Infectious Disease,</institution><institution>University of Texas Medical Branch,</institution></institution-wrap>Galveston, TX USA</aff></contrib-group><pub-date pub-type="epub"><day>14</day><month>9</month><year>2019</year></pub-date><pub-date pub-type="collection"><year>2020</year></pub-date><volume>2099</volume><fpage>99</fpage><lpage>106</lpage><permissions><copyright-statement>© Springer Science+Business Media, LLC, part of Springer Nature 2020</copyright-statement><license><license-p>This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.</license-p></license></permissions><abstract id="Abs1"><p id="Par1">Since the emergence of the Middle East respiratory syndrome-coronavirus (MERS-CoV) in 2012, more than 2280 confirmed human infections and 800 associated deaths had been reported to the World Health Organization. MERS-CoV is a single-stranded RNA virus that belongs to the <italic>Coronaviridae</italic> family. MERS-CoV infection leads to a variety of clinical outcomes in humans ranging from asymptomatic and mild infection to severe acute lung injury and multi-organ failure and death. To study the pathogenesis of MERS-CoV infection and development of medical countermeasures (MCMs) for MERS, a number of genetically modified mouse models have been developed, including various versions of transgenic mice expressing the human DPP4 viral receptor. Tracking and quantifying viral infection, among others, in permissive hosts is a key endpoint for studying MERS pathogenesis and evaluating the efficacy of selected MCMs developed for MERS. In addition to quantifying infectious progeny virus which requires high-containment biosafety level (BSL)-3 laboratory, here we outlined an established real-time quantitative RT-PCR (RT-qPCR)-based procedure to unequivocally quantify MERS-CoV-specific RNAs within the lungs of infected human DPP4 (hDPP4, transgenic (hDPP4 Tg) mice under a standard BSL-2 laboratory.</p></abstract><kwd-group xml:lang="en"><title>Key words</title><kwd>MERS-CoV</kwd><kwd>RT-qPCR</kwd><kwd>Animal models</kwd></kwd-group><custom-meta-group><custom-meta><meta-name>issue-copyright-statement</meta-name><meta-value>© Springer Science+Business Media, LLC, part of Springer Nature 2020</meta-value></custom-meta></custom-meta-group></article-meta></front></pmc><affiliations><list></list><tree></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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