Development of Dual TaqMan Based One-Step rRT-PCR Assay Panel for Rapid and Accurate Diagnostic Test of MERS-CoV: A Novel Human Coronavirus, Ahead of Hajj Pilgrimage
Identifieur interne : 000D74 ( Pmc/Curation ); précédent : 000D73; suivant : 000D75Development of Dual TaqMan Based One-Step rRT-PCR Assay Panel for Rapid and Accurate Diagnostic Test of MERS-CoV: A Novel Human Coronavirus, Ahead of Hajj Pilgrimage
Auteurs : Mohammad Sadegh Hashemzadeh ; Rahimeh Rasouli ; Bentolhoda Zahraei ; Morteza Izadi ; Mahdi Tat ; Seyed Hassan Saadat ; Mohammad Najarasl ; Behzad Khansari Nejad ; Ruhollah DorostkarSource :
- Iranian Red Crescent Medical Journal [ 2074-1804 ] ; 2016.
Abstract
Coronaviruses (CoVs) are large ribonucleic acid (RNA) viruses causing primarily respiratory disease in humans. A novel human coronavirus, subsequently named middle east respiratory syndrome coronavirus (MERS-CoV), was first reported in Saudi Arabia in September of 2012. With increasing numbers of infections and deaths from MERS-CoV, development of a rapid and reliable kit was crucial to prevent further spread of MERS-CoV.
In this study, we present two real-time reverse-transcription polymerase chain reaction (rRT-PCR) assays for in-house rapid and sensitive diagnostic testing of MERS-CoV, detecting the regions upstream of the envelope gene (upE) and open reading frame (ORF) 1b, respectively, for initial screening and final confirmation of MERS-CoV infection, as recommended by the world health organization (WHO).
In this experimental study, acquiring patient samples was difficult; thus, according to WHO recommendations and standard protocols, we synthesized RNA sequences of upE and ORF1b genes as the template signatures and TaqMan based-diagnostic rRT-PCR assays were carried out using these synthetic genes for detection of MERS-CoV. In this research, we also inaugurated a cell-free system to transcribe these RNA sequences using the DNA templates synthesized.
The upE and ORF1b based one-step rRT-PCR assays were optimized by testing several times via different synthetic RNAs, and validation results were highly successful. The sensitivity obtained for upE was fewer than ten copies of RNA template per reaction and for ORF1b was 50 or fewer copies per reaction.
This study showed that the developed rRT-PCR assays are rapid, reliable, reproducible, specific, sensitive, and simple tools for detection of MERS-CoV. Finally, a kit consisting of two assay signatures and controls was assembled, which can be distributed to public health laboratories in Iran to support international MERS-CoV surveillance and public health response.
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DOI: 10.5812/ircmj.23874
PubMed: 28191331
PubMed Central: 5292137
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Mohammad Sadegh Hashemzadeh<affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff96433">Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff96433">Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff96434">Department of Microbiology and Immunology, Arak University of Medical Sciences, Arak, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation>Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development of Dual TaqMan Based One-Step rRT-PCR Assay Panel for Rapid and Accurate Diagnostic Test of MERS-CoV: A Novel Human Coronavirus, Ahead of Hajj Pilgrimage</title><author><name sortKey="Hashemzadeh, Mohammad Sadegh" sort="Hashemzadeh, Mohammad Sadegh" uniqKey="Hashemzadeh M" first="Mohammad Sadegh" last="Hashemzadeh">Mohammad Sadegh Hashemzadeh</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Rasouli, Rahimeh" sort="Rasouli, Rahimeh" uniqKey="Rasouli R" first="Rahimeh" last="Rasouli">Rahimeh Rasouli</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Zahraei, Bentolhoda" sort="Zahraei, Bentolhoda" uniqKey="Zahraei B" first="Bentolhoda" last="Zahraei">Bentolhoda Zahraei</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Izadi, Morteza" sort="Izadi, Morteza" uniqKey="Izadi M" first="Morteza" last="Izadi">Morteza Izadi</name><affiliation><nlm:aff id="aff96433">Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Tat, Mahdi" sort="Tat, Mahdi" uniqKey="Tat M" first="Mahdi" last="Tat">Mahdi Tat</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Saadat, Seyed Hassan" sort="Saadat, Seyed Hassan" uniqKey="Saadat S" first="Seyed Hassan" last="Saadat">Seyed Hassan Saadat</name><affiliation><nlm:aff id="aff96433">Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Najarasl, Mohammad" sort="Najarasl, Mohammad" uniqKey="Najarasl M" first="Mohammad" last="Najarasl">Mohammad Najarasl</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Khansari Nejad, Behzad" sort="Khansari Nejad, Behzad" uniqKey="Khansari Nejad B" first="Behzad" last="Khansari Nejad">Behzad Khansari Nejad</name><affiliation><nlm:aff id="aff96434">Department of Microbiology and Immunology, Arak University of Medical Sciences, Arak, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Dorostkar, Ruhollah" sort="Dorostkar, Ruhollah" uniqKey="Dorostkar R" first="Ruhollah" last="Dorostkar">Ruhollah Dorostkar</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">28191331</idno><idno type="pmc">5292137</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5292137</idno><idno type="RBID">PMC:5292137</idno><idno type="doi">10.5812/ircmj.23874</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000D74</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000D74</idno><idno type="wicri:Area/Pmc/Curation">000D74</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000D74</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Development of Dual TaqMan Based One-Step rRT-PCR Assay Panel for Rapid and Accurate Diagnostic Test of MERS-CoV: A Novel Human Coronavirus, Ahead of Hajj Pilgrimage</title><author><name sortKey="Hashemzadeh, Mohammad Sadegh" sort="Hashemzadeh, Mohammad Sadegh" uniqKey="Hashemzadeh M" first="Mohammad Sadegh" last="Hashemzadeh">Mohammad Sadegh Hashemzadeh</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Rasouli, Rahimeh" sort="Rasouli, Rahimeh" uniqKey="Rasouli R" first="Rahimeh" last="Rasouli">Rahimeh Rasouli</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Zahraei, Bentolhoda" sort="Zahraei, Bentolhoda" uniqKey="Zahraei B" first="Bentolhoda" last="Zahraei">Bentolhoda Zahraei</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Izadi, Morteza" sort="Izadi, Morteza" uniqKey="Izadi M" first="Morteza" last="Izadi">Morteza Izadi</name><affiliation><nlm:aff id="aff96433">Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Tat, Mahdi" sort="Tat, Mahdi" uniqKey="Tat M" first="Mahdi" last="Tat">Mahdi Tat</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Saadat, Seyed Hassan" sort="Saadat, Seyed Hassan" uniqKey="Saadat S" first="Seyed Hassan" last="Saadat">Seyed Hassan Saadat</name><affiliation><nlm:aff id="aff96433">Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Najarasl, Mohammad" sort="Najarasl, Mohammad" uniqKey="Najarasl M" first="Mohammad" last="Najarasl">Mohammad Najarasl</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Khansari Nejad, Behzad" sort="Khansari Nejad, Behzad" uniqKey="Khansari Nejad B" first="Behzad" last="Khansari Nejad">Behzad Khansari Nejad</name><affiliation><nlm:aff id="aff96434">Department of Microbiology and Immunology, Arak University of Medical Sciences, Arak, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Dorostkar, Ruhollah" sort="Dorostkar, Ruhollah" uniqKey="Dorostkar R" first="Ruhollah" last="Dorostkar">Ruhollah Dorostkar</name><affiliation><nlm:aff id="aff96432">Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author></analytic><series><title level="j">Iranian Red Crescent Medical Journal</title><idno type="ISSN">2074-1804</idno><idno type="eISSN">2074-1812</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>Coronaviruses (CoVs) are large ribonucleic acid (RNA) viruses causing primarily respiratory disease in humans. A novel human coronavirus, subsequently named middle east respiratory syndrome coronavirus (MERS-CoV), was first reported in Saudi Arabia in September of 2012. With increasing numbers of infections and deaths from MERS-CoV, development of a rapid and reliable kit was crucial to prevent further spread of MERS-CoV.</p></sec><sec><title>Objectives</title><p>In this study, we present two real-time reverse-transcription polymerase chain reaction (rRT-PCR) assays for in-house rapid and sensitive diagnostic testing of MERS-CoV, detecting the regions upstream of the envelope gene (upE) and open reading frame (ORF) 1b, respectively, for initial screening and final confirmation of MERS-CoV infection, as recommended by the world health organization (WHO).</p></sec><sec><title>Materials and Methods</title><p>In this experimental study, acquiring patient samples was difficult; thus, according to WHO recommendations and standard protocols, we synthesized RNA sequences of upE and ORF1b genes as the template signatures and TaqMan based-diagnostic rRT-PCR assays were carried out using these synthetic genes for detection of MERS-CoV. In this research, we also inaugurated a cell-free system to transcribe these RNA sequences using the DNA templates synthesized.</p></sec><sec><title>Results</title><p>The upE and ORF1b based one-step rRT-PCR assays were optimized by testing several times via different synthetic RNAs, and validation results were highly successful. The sensitivity obtained for upE was fewer than ten copies of RNA template per reaction and for ORF1b was 50 or fewer copies per reaction.</p></sec><sec><title>Conclusions</title><p>This study showed that the developed rRT-PCR assays are rapid, reliable, reproducible, specific, sensitive, and simple tools for detection of MERS-CoV. Finally, a kit consisting of two assay signatures and controls was assembled, which can be distributed to public health laboratories in Iran to support international MERS-CoV surveillance and public health response.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="King, Amq" uniqKey="King A">AMQ King</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, Tm" uniqKey="Bestebroer T">TM Bestebroer</name></author><author><name sortKey="Osterhaus, Ad" uniqKey="Osterhaus A">AD Osterhaus</name></author><author><name sortKey="Fouchier, Ra" uniqKey="Fouchier R">RA Fouchier</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Memish, Za" uniqKey="Memish Z">ZA Memish</name></author><author><name sortKey="Zumla, Ai" uniqKey="Zumla A">AI Zumla</name></author><author><name sortKey="Al Hakeem, Rf" uniqKey="Al Hakeem R">RF Al-Hakeem</name></author><author><name sortKey="Al Rabeeah, Aa" uniqKey="Al Rabeeah A">AA Al-Rabeeah</name></author><author><name sortKey="Stephens, Gm" uniqKey="Stephens G">GM Stephens</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Assiri, A" uniqKey="Assiri A">A Assiri</name></author><author><name sortKey="Al Tawfiq, Ja" uniqKey="Al Tawfiq J">JA Al-Tawfiq</name></author><author><name sortKey="Al Rabeeah, Aa" uniqKey="Al Rabeeah A">AA Al-Rabeeah</name></author><author><name sortKey="Al Rabiah, Fa" uniqKey="Al Rabiah F">FA Al-Rabiah</name></author><author><name sortKey="Al Hajjar, S" uniqKey="Al Hajjar S">S Al-Hajjar</name></author><author><name sortKey="Al Barrak, A" uniqKey="Al Barrak A">A Al-Barrak</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Balkhair, A" uniqKey="Balkhair A">A 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Iran Red Crescent Med J</journal-id><journal-id journal-id-type="iso-abbrev">Iran Red Crescent Med J</journal-id><journal-id journal-id-type="doi">10.5812/ircmj</journal-id><journal-id journal-id-type="publisher-id">Kowsar</journal-id><journal-title-group><journal-title>Iranian Red Crescent Medical Journal</journal-title></journal-title-group><issn pub-type="ppub">2074-1804</issn><issn pub-type="epub">2074-1812</issn><publisher><publisher-name>Kowsar</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">28191331</article-id><article-id pub-id-type="pmc">5292137</article-id><article-id pub-id-type="doi">10.5812/ircmj.23874</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title>Development of Dual TaqMan Based One-Step rRT-PCR Assay Panel for Rapid and Accurate Diagnostic Test of MERS-CoV: A Novel Human Coronavirus, Ahead of Hajj Pilgrimage</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Hashemzadeh</surname><given-names>Mohammad Sadegh</given-names></name><xref ref-type="aff" rid="aff96432">1</xref></contrib><contrib contrib-type="author"><name><surname>Rasouli</surname><given-names>Rahimeh</given-names></name><xref ref-type="aff" rid="aff96432">1</xref></contrib><contrib contrib-type="author"><name><surname>Zahraei</surname><given-names>Bentolhoda</given-names></name><xref ref-type="aff" rid="aff96432">1</xref></contrib><contrib contrib-type="author"><name><surname>Izadi</surname><given-names>Morteza</given-names></name><xref ref-type="aff" rid="aff96433">2</xref></contrib><contrib contrib-type="author"><name><surname>Tat</surname><given-names>Mahdi</given-names></name><xref ref-type="aff" rid="aff96432">1</xref></contrib><contrib contrib-type="author"><name><surname>Saadat</surname><given-names>Seyed Hassan</given-names></name><xref ref-type="aff" rid="aff96433">2</xref></contrib><contrib contrib-type="author"><name><surname>Najarasl</surname><given-names>Mohammad</given-names></name><xref ref-type="aff" rid="aff96432">1</xref></contrib><contrib contrib-type="author"><name><surname>Khansari Nejad</surname><given-names>Behzad</given-names></name><xref ref-type="aff" rid="aff96434">3</xref></contrib><contrib contrib-type="author"><name><surname>Dorostkar</surname><given-names>Ruhollah</given-names></name><xref ref-type="aff" rid="aff96432">1</xref><xref ref-type="corresp" rid="cor96435">*</xref></contrib></contrib-group><aff id="aff96432"><label>1</label>Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</aff><aff id="aff96433"><label>2</label>Health Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran</aff><aff id="aff96434"><label>3</label>Department of Microbiology and Immunology, Arak University of Medical Sciences, Arak, IR Iran</aff><author-notes><corresp id="cor96435"><label>*</label>Corresponding Author: Ruhollah Dorostkar, Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran. Tel/Fax: +98-2188617711-16, E-mail: <email>R.dorost@yahoo.com</email></corresp></author-notes><pub-date pub-type="epub"><day>21</day><month>6</month><year>2016</year></pub-date><pub-date pub-type="collection"><month>11</month><year>2016</year></pub-date><volume>18</volume><issue>11</issue><elocation-id>e23874</elocation-id><history><date date-type="received"><day>30</day><month>9</month><year>2014</year></date><date date-type="rev-recd"><day>28</day><month>11</month><year>2014</year></date><date date-type="accepted"><day>05</day><month>4</month><year>2015</year></date></history><permissions><copyright-statement>Copyright © 2016, Iranian Red Crescent Medical Journal</copyright-statement><copyright-year>2016</copyright-year><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc/4.0/"><license-p><pmc-comment>CREATIVE COMMONS</pmc-comment>This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by-nc/4.0/">http://creativecommons.org/licenses/by-nc/4.0/</ext-link>) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.</license-p></license></permissions><abstract><sec><title>Background</title><p>Coronaviruses (CoVs) are large ribonucleic acid (RNA) viruses causing primarily respiratory disease in humans. A novel human coronavirus, subsequently named middle east respiratory syndrome coronavirus (MERS-CoV), was first reported in Saudi Arabia in September of 2012. With increasing numbers of infections and deaths from MERS-CoV, development of a rapid and reliable kit was crucial to prevent further spread of MERS-CoV.</p></sec><sec><title>Objectives</title><p>In this study, we present two real-time reverse-transcription polymerase chain reaction (rRT-PCR) assays for in-house rapid and sensitive diagnostic testing of MERS-CoV, detecting the regions upstream of the envelope gene (upE) and open reading frame (ORF) 1b, respectively, for initial screening and final confirmation of MERS-CoV infection, as recommended by the world health organization (WHO).</p></sec><sec><title>Materials and Methods</title><p>In this experimental study, acquiring patient samples was difficult; thus, according to WHO recommendations and standard protocols, we synthesized RNA sequences of upE and ORF1b genes as the template signatures and TaqMan based-diagnostic rRT-PCR assays were carried out using these synthetic genes for detection of MERS-CoV. In this research, we also inaugurated a cell-free system to transcribe these RNA sequences using the DNA templates synthesized.</p></sec><sec><title>Results</title><p>The upE and ORF1b based one-step rRT-PCR assays were optimized by testing several times via different synthetic RNAs, and validation results were highly successful. The sensitivity obtained for upE was fewer than ten copies of RNA template per reaction and for ORF1b was 50 or fewer copies per reaction.</p></sec><sec><title>Conclusions</title><p>This study showed that the developed rRT-PCR assays are rapid, reliable, reproducible, specific, sensitive, and simple tools for detection of MERS-CoV. Finally, a kit consisting of two assay signatures and controls was assembled, which can be distributed to public health laboratories in Iran to support international MERS-CoV surveillance and public health response.</p></sec></abstract><kwd-group kwd-group-type="author"><kwd>Hajj Pilgrimage</kwd><kwd>MERS-CoV</kwd><kwd>Diagnosis</kwd><kwd>Real-Time RT-PCR</kwd><kwd>upE</kwd><kwd>ORF1b</kwd></kwd-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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