Quantification of the Middle East Respiratory Syndrome-Coronavirus RNA in Tissues by Quantitative Real-Time RT-PCR
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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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<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>
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<journal-subtitle>Methods and Protocols </journal-subtitle>
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<isbn publication-format="print">978-1-0716-0210-2</isbn>
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<contrib-group content-type="book editors"><contrib contrib-type="editor"><name><surname>Vijay</surname>
<given-names>Rahul</given-names>
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<address><email>rahul-vijay@uiowa.edu</email>
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<institution>Department of Microbiology and Immunology,</institution>
<institution>University of Iowa,</institution>
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Iowa City, IA USA</aff>
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<contrib-group><contrib contrib-type="author"><name><surname>Algaissi</surname>
<given-names>Abdullah</given-names>
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<given-names>Anurodh S.</given-names>
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<contrib contrib-type="author" corresp="yes"><name><surname>Hashem</surname>
<given-names>Anwar M.</given-names>
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<address><email>amhashem@kau.edu.sa</email>
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<contrib contrib-type="author" corresp="yes"><name><surname>Tseng</surname>
<given-names>Chien-Te K.</given-names>
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<address><email>sktseng@utmb.edu</email>
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Jazan, Saudi Arabia</aff>
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Jeddah, Saudi Arabia</aff>
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Galveston, TX USA</aff>
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<pub-date pub-type="epub"><day>14</day>
<month>9</month>
<year>2019</year>
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<pub-date pub-type="collection"><year>2020</year>
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<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>
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<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>
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<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>
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