Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA
Identifieur interne : 000A52 ( Pmc/Curation ); précédent : 000A51; suivant : 000A53Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA
Auteurs : Hee Jae Huh [Corée du Sud] ; Ji-Youn Kim [Corée du Sud] ; Hyeon Jeong Kwon [Corée du Sud] ; Sun Ae Yun [Corée du Sud] ; Myoung-Keun Lee [Corée du Sud] ; Chang-Seok Ki [Corée du Sud] ; Nam Yong Lee [Corée du Sud] ; Jong-Won Kim [Corée du Sud]Source :
- Annals of Laboratory Medicine [ 2234-3806 ] ; 2017.
Abstract
Molecular detection of Middle East respiratory syndrome coronavirus (MERS-CoV) using real-time reverse transcription (rRT)-PCR assays is the method of choice for diagnosis of MERS. We evaluated the performance of the PowerChek MERS (upE & ORF1a) real-time PCR Kit (PowerChek MERS assay; Kogene Biotech, Korea) a one-step rRT-PCR assay for the qualitative detection of MERS-CoV.
We evaluated PowerChek MERS assay performance in comparison with nested RT-PCR and sequencing of the RNA-dependent RNA polymerase (RdRp) and N genes. To evaluate diagnostic sensitivity and specificity, 100 clinical specimens (50 positive and 50 negative for MERS-CoV) were simultaneously tested by using the PowerChek MERS and sequencing assays. Assay performance, including limit of detection and precision, was evaluated in vitro by using MERS-CoV RNA transcripts. Analytical specificity was evaluated with a diverse collection of 16 respiratory virus–positive clinical specimens and 14 respiratory bacterial isolates.
The 95% limits of detection of the PowerChek MERS assay for the upE and the open rading frame (ORF)1a were 16.2 copies/µL and 8.2 copies/µL, respectively. No cross-reactivity was observed. The diagnostic sensitivity and specificity of the PowerChek MERS assay were both 100% (95% confidence interval, 91.1–100%).
The PowerChek MERS assay is a straightforward and accurate assay for detecting MERS-CoV RNA. The assay will be a useful tool for the rapid diagnosis of MERS and could prove especially important for MERS outbreak control.
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DOI: 10.3343/alm.2017.37.6.494
PubMed: 28840986
PubMed Central: 5587821
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA</title><author><name sortKey="Huh, Hee Jae" sort="Huh, Hee Jae" uniqKey="Huh H" first="Hee Jae" last="Huh">Hee Jae Huh</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Kim, Ji Youn" sort="Kim, Ji Youn" uniqKey="Kim J" first="Ji-Youn" last="Kim">Ji-Youn Kim</name><affiliation wicri:level="1"><nlm:aff id="A2">Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Kwon, Hyeon Jeong" sort="Kwon, Hyeon Jeong" uniqKey="Kwon H" first="Hyeon Jeong" last="Kwon">Hyeon Jeong Kwon</name><affiliation wicri:level="1"><nlm:aff id="A2">Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Yun, Sun Ae" sort="Yun, Sun Ae" uniqKey="Yun S" first="Sun Ae" last="Yun">Sun Ae Yun</name><affiliation wicri:level="1"><nlm:aff id="A2">Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Lee, Myoung Keun" sort="Lee, Myoung Keun" uniqKey="Lee M" first="Myoung-Keun" last="Lee">Myoung-Keun Lee</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Ki, Chang Seok" sort="Ki, Chang Seok" uniqKey="Ki C" first="Chang-Seok" last="Ki">Chang-Seok Ki</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Lee, Nam Yong" sort="Lee, Nam Yong" uniqKey="Lee N" first="Nam Yong" last="Lee">Nam Yong Lee</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Kim, Jong Won" sort="Kim, Jong Won" uniqKey="Kim J" first="Jong-Won" last="Kim">Jong-Won Kim</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">28840986</idno><idno type="pmc">5587821</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5587821</idno><idno type="RBID">PMC:5587821</idno><idno type="doi">10.3343/alm.2017.37.6.494</idno><date when="2017">2017</date><idno type="wicri:Area/Pmc/Corpus">000A52</idno><idno type="wicri:explorRef" 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Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Kim, Ji Youn" sort="Kim, Ji Youn" uniqKey="Kim J" first="Ji-Youn" last="Kim">Ji-Youn Kim</name><affiliation wicri:level="1"><nlm:aff id="A2">Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Kwon, Hyeon Jeong" sort="Kwon, Hyeon Jeong" uniqKey="Kwon H" first="Hyeon Jeong" last="Kwon">Hyeon Jeong Kwon</name><affiliation wicri:level="1"><nlm:aff id="A2">Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Yun, Sun Ae" sort="Yun, Sun Ae" uniqKey="Yun S" first="Sun Ae" last="Yun">Sun Ae Yun</name><affiliation wicri:level="1"><nlm:aff id="A2">Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Lee, Myoung Keun" sort="Lee, Myoung Keun" uniqKey="Lee M" first="Myoung-Keun" last="Lee">Myoung-Keun Lee</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Ki, Chang Seok" sort="Ki, Chang Seok" uniqKey="Ki C" first="Chang-Seok" last="Ki">Chang-Seok Ki</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Lee, Nam Yong" sort="Lee, Nam Yong" uniqKey="Lee N" first="Nam Yong" last="Lee">Nam Yong Lee</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Kim, Jong Won" sort="Kim, Jong Won" uniqKey="Kim J" first="Jong-Won" last="Kim">Jong-Won Kim</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea></affiliation></author></analytic><series><title level="j">Annals of Laboratory Medicine</title><idno type="ISSN">2234-3806</idno><idno type="eISSN">2234-3814</idno><imprint><date when="2017">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>Molecular detection of Middle East respiratory syndrome coronavirus (MERS-CoV) using real-time reverse transcription (rRT)-PCR assays is the method of choice for diagnosis of MERS. We evaluated the performance of the PowerChek MERS (upE & ORF1a) real-time PCR Kit (PowerChek MERS assay; Kogene Biotech, Korea) a one-step rRT-PCR assay for the qualitative detection of MERS-CoV.</p></sec><sec><title>Methods</title><p>We evaluated PowerChek MERS assay performance in comparison with nested RT-PCR and sequencing of the RNA-dependent RNA polymerase (RdRp) and N genes. To evaluate diagnostic sensitivity and specificity, 100 clinical specimens (50 positive and 50 negative for MERS-CoV) were simultaneously tested by using the PowerChek MERS and sequencing assays. Assay performance, including limit of detection and precision, was evaluated in vitro by using MERS-CoV RNA transcripts. Analytical specificity was evaluated with a diverse collection of 16 respiratory virus–positive clinical specimens and 14 respiratory bacterial isolates.</p></sec><sec><title>Results</title><p>The 95% limits of detection of the PowerChek MERS assay for the upE and the open rading frame (ORF)1a were 16.2 copies/µL and 8.2 copies/µL, respectively. No cross-reactivity was observed. The diagnostic sensitivity and specificity of the PowerChek MERS assay were both 100% (95% confidence interval, 91.1–100%).</p></sec><sec><title>Conclusions</title><p>The PowerChek MERS assay is a straightforward and accurate assay for detecting MERS-CoV RNA. The assay will be a useful tool for the rapid diagnosis of MERS and could prove especially important for MERS outbreak control.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Chan, Jf" uniqKey="Chan J">JF Chan</name></author><author><name sortKey="Lau, Sk" uniqKey="Lau S">SK Lau</name></author><author><name sortKey="To, Kk" uniqKey="To K">KK To</name></author><author><name sortKey="Cheng, Vc" uniqKey="Cheng V">VC Cheng</name></author><author><name sortKey="Woo, Pc" uniqKey="Woo P">PC Woo</name></author><author><name sortKey="Yuen, Ky" uniqKey="Yuen K">KY Yuen</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Corman, Vm" uniqKey="Corman V">VM Corman</name></author><author><name sortKey="Eckerle, I" uniqKey="Eckerle I">I Eckerle</name></author><author><name sortKey="Bleicker, T" uniqKey="Bleicker T">T Bleicker</name></author><author><name sortKey="Zaki, A" 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Ann Lab Med</journal-id><journal-id journal-id-type="iso-abbrev">Ann Lab Med</journal-id><journal-id journal-id-type="publisher-id">ALM</journal-id><journal-title-group><journal-title>Annals of Laboratory Medicine</journal-title></journal-title-group><issn pub-type="ppub">2234-3806</issn><issn pub-type="epub">2234-3814</issn><publisher><publisher-name>The Korean Society for Laboratory Medicine</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">28840986</article-id><article-id pub-id-type="pmc">5587821</article-id><article-id pub-id-type="doi">10.3343/alm.2017.37.6.494</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject><subj-group subj-group-type="subheading"><subject>Clinical Microbiology</subject></subj-group></subj-group></article-categories><title-group><article-title>Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Huh</surname><given-names>Hee Jae</given-names></name><degrees>M.D.</degrees><xref ref-type="aff" rid="A1">1</xref></contrib><contrib contrib-type="author"><name><surname>Kim</surname><given-names>Ji-Youn</given-names></name><degrees>M.T.</degrees><xref ref-type="aff" rid="A2">2</xref></contrib><contrib contrib-type="author"><name><surname>Kwon</surname><given-names>Hyeon Jeong</given-names></name><degrees>M.T.</degrees><xref ref-type="aff" rid="A2">2</xref></contrib><contrib contrib-type="author"><name><surname>Yun</surname><given-names>Sun Ae</given-names></name><degrees>M.T.</degrees><xref ref-type="aff" rid="A2">2</xref></contrib><contrib contrib-type="author"><name><surname>Lee</surname><given-names>Myoung-Keun</given-names></name><degrees>M.T.</degrees><xref ref-type="aff" rid="A1">1</xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Ki</surname><given-names>Chang-Seok</given-names></name><degrees>M.D.</degrees><xref ref-type="aff" rid="A1">1</xref></contrib><contrib contrib-type="author"><name><surname>Lee</surname><given-names>Nam Yong</given-names></name><degrees>M.D.</degrees><xref ref-type="aff" rid="A1">1</xref></contrib><contrib contrib-type="author"><name><surname>Kim</surname><given-names>Jong-Won</given-names></name><degrees>M.D.</degrees><xref ref-type="aff" rid="A1">1</xref></contrib></contrib-group><aff id="A1"><label>1</label>Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</aff><aff id="A2"><label>2</label>Center for Clinical Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Korea.</aff><author-notes><corresp>Corresponding author: Chang-Seok Ki. Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, 81 Irwon-ro, Gangnam-gu, Seoul 06351, Korea. Tel: +82-2-3410-2709, Fax: +82-2-3410-2719, <email>changski@skku.edu</email></corresp></author-notes><pub-date pub-type="ppub"><month>11</month><year>2017</year></pub-date><pub-date pub-type="epub"><day>16</day><month>8</month><year>2017</year></pub-date><volume>37</volume><issue>6</issue><fpage>494</fpage><lpage>498</lpage><history><date date-type="received"><day>03</day><month>2</month><year>2017</year></date><date date-type="rev-recd"><day>26</day><month>3</month><year>2017</year></date><date date-type="accepted"><day>12</day><month>7</month><year>2017</year></date></history><permissions><copyright-statement>© The Korean Society for Laboratory Medicine</copyright-statement><copyright-year>2017</copyright-year><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc/4.0/"><license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial 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 unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><abstract><sec><title>Background</title><p>Molecular detection of Middle East respiratory syndrome coronavirus (MERS-CoV) using real-time reverse transcription (rRT)-PCR assays is the method of choice for diagnosis of MERS. We evaluated the performance of the PowerChek MERS (upE & ORF1a) real-time PCR Kit (PowerChek MERS assay; Kogene Biotech, Korea) a one-step rRT-PCR assay for the qualitative detection of MERS-CoV.</p></sec><sec><title>Methods</title><p>We evaluated PowerChek MERS assay performance in comparison with nested RT-PCR and sequencing of the RNA-dependent RNA polymerase (RdRp) and N genes. To evaluate diagnostic sensitivity and specificity, 100 clinical specimens (50 positive and 50 negative for MERS-CoV) were simultaneously tested by using the PowerChek MERS and sequencing assays. Assay performance, including limit of detection and precision, was evaluated in vitro by using MERS-CoV RNA transcripts. Analytical specificity was evaluated with a diverse collection of 16 respiratory virus–positive clinical specimens and 14 respiratory bacterial isolates.</p></sec><sec><title>Results</title><p>The 95% limits of detection of the PowerChek MERS assay for the upE and the open rading frame (ORF)1a were 16.2 copies/µL and 8.2 copies/µL, respectively. No cross-reactivity was observed. The diagnostic sensitivity and specificity of the PowerChek MERS assay were both 100% (95% confidence interval, 91.1–100%).</p></sec><sec><title>Conclusions</title><p>The PowerChek MERS assay is a straightforward and accurate assay for detecting MERS-CoV RNA. The assay will be a useful tool for the rapid diagnosis of MERS and could prove especially important for MERS outbreak control.</p></sec></abstract><kwd-group><kwd>MERS-CoV</kwd><kwd>Real-time reverse-transcription PCR</kwd><kwd>Performance</kwd></kwd-group><funding-group><award-group><funding-source><institution-wrap><institution>Korea Centers for Disease Control and Prevention</institution><institution-id institution-id-type="CrossRef">http://dx.doi.org/10.13039/501100003669</institution-id></institution-wrap></funding-source><award-id>HD15A3232</award-id></award-group></funding-group></article-meta></front><floats-group><table-wrap id="T1" orientation="portrait" position="float"><label>Table 1</label><caption><title>Analytical specificity of the PowerChek MERS (upE and ORF1a) real-time PCR Kit</title></caption><alternatives><graphic xlink:href="alm-37-494-i001"></graphic><table frame="hsides" rules="rows"><col width="35%" span="1"></col><col width="45%" span="1"></col><col width="20%" span="1"></col><thead><tr><th valign="middle" align="left" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">Microorganism</th><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">Source</th><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">PowerChek MERS assay</th></tr></thead><tbody><tr><td valign="top" align="left" rowspan="1" colspan="1">Viruses</td><td valign="top" align="left" rowspan="1" colspan="1"></td><td valign="top" align="left" rowspan="1" colspan="1"></td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Adenovirus</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Bocavirus</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Coronavirus 229E</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Coronavirus OC43</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Coronavirus HKU1</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Coronavirus NL63</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Human enterovirus</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Human metapneumovirus</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Human rhinovirus</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Influenza A</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Influenza B</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Parainfluenza 1</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Parainfluenza 2</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Parainfluenza 3</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Respiratory syncytial virus A</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"> Respiratory syncytial virus B</td><td valign="top" align="left" rowspan="1" colspan="1">Clinical specimen<sup>*</sup></td><td valign="top" align="left" rowspan="1" colspan="1">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Bacteria</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"></td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"></td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Acinetobacter baumannii</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 19606)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Escherichia coli</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 25922)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Haemophilus influenza</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 9007)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Klebsiella pneumoniae</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 900603)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Legionella pneumophila</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 33156)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Mycoplasma pneumoniae</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Clinical specimen<sup>†</sup></td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Mycobacterium tuberculosis</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 27294)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Mycobacterium avium</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (KMRC 00136-41011)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Mycobacterium abscessus</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 19977)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Pseudomonas aeruginosa</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 27853)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Staphylococcus aureus</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 25923)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Staphylococcus epidermidis</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 12228)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Streptococcus pneumoniae</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 49619)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)"> Streptococcus pyogenes</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Reference strain (ATCC 19615)</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td></tr></tbody></table></alternatives><table-wrap-foot><fn><p><sup>*</sup>Specimens that yielded positive results in two respiratory virus panel assays: the AdvanSure RV real-time PCR Kit (LG chem, Seoul, Korea) and the Allplex Respiratory Panel 1, 2, and 3 (Seegene, Seoul, Korea); <sup>†</sup>Specimens confirmed by sequencing analysis.</p><p>Abbreviation: KMRC, Korea Mycobacterium Resource Center.</p></fn></table-wrap-foot></table-wrap><table-wrap id="T2" orientation="portrait" position="float"><label>Table 2</label><caption><title>Analytical sensitivity of the PowerChek MERS (upE and ORF1a) real-time PCR Kit</title></caption><alternatives><graphic xlink:href="alm-37-494-i002"></graphic><table frame="hsides" rules="rows"><col width="40%" span="1"></col><col width="30%" span="1"></col><col width="30%" span="1"></col><thead><tr><th valign="middle" align="left" rowspan="2" colspan="1" style="background-color:rgb(218,227,244)">Concentration (copies/µL)</th><th valign="middle" align="center" rowspan="1" colspan="2" style="background-color:rgb(218,227,244)">N of detections/N of replicates (%)</th></tr><tr><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">upE</th><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">ORF1a</th></tr></thead><tbody><tr><td valign="top" align="left" rowspan="1" colspan="1">100</td><td valign="top" align="center" rowspan="1" colspan="1">20/20 (100)</td><td valign="top" align="center" rowspan="1" colspan="1">20/20 (100)</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">50</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">20/20 (100)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">20/20 (100)</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1">25</td><td valign="top" align="center" rowspan="1" colspan="1">20/20 (100)</td><td valign="top" align="center" rowspan="1" colspan="1">20/20 (100)</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">12.5</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">17/20 (85)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">20/20 (100)</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1">10</td><td valign="top" align="center" rowspan="1" colspan="1">18/20 (90)</td><td valign="top" align="center" rowspan="1" colspan="1">19/20 (95)</td></tr></tbody></table></alternatives></table-wrap><table-wrap id="T3" orientation="portrait" position="float"><label>Table 3</label><caption><title>Precision of the PowerChek MERS (upE and ORF1a) real-time PCR Kit</title></caption><alternatives><graphic xlink:href="alm-37-494-i003"></graphic><table frame="hsides" rules="rows"><col width="9.12%" span="1"></col><col width="9.12%" span="1"></col><col width="8.95%" span="1"></col><col width="8.95%" span="1"></col><col width="8.95%" span="1"></col><col width="8.95%" span="1"></col><col width="8.95%" span="1"></col><col width="8.95%" span="1"></col><thead><tr><th valign="middle" align="left" rowspan="3" colspan="1" style="background-color:rgb(218,227,244)">Target</th><th valign="middle" align="center" rowspan="3" colspan="1" style="background-color:rgb(218,227,244)">Concentration (copies/µL)</th><th valign="middle" align="center" rowspan="1" colspan="6" style="background-color:rgb(218,227,244)">Mean Ct (% coefficient of variation)</th></tr><tr><th valign="middle" align="center" rowspan="1" colspan="2" style="background-color:rgb(218,227,244)">Reproducibility</th><th valign="middle" align="center" rowspan="2" colspan="1" style="background-color:rgb(218,227,244)">Repeatability</th><th valign="middle" align="center" rowspan="1" colspan="3" style="background-color:rgb(218,227,244)">Lot-to-lot variation</th></tr><tr><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">Site 1</th><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">Site 2</th><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">Lot 1</th><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">Lot 2</th><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">Lot 3</th></tr></thead><tbody><tr><td valign="middle" align="left" rowspan="3" colspan="1">upE</td><td valign="middle" align="right" rowspan="1" colspan="1">50</td><td valign="middle" align="center" rowspan="1" colspan="1">34.40 (2.10)</td><td valign="middle" align="center" rowspan="1" colspan="1">34.40 (1.32)</td><td valign="middle" align="center" rowspan="1" colspan="1">34.74 (2.11)</td><td valign="middle" align="center" rowspan="1" colspan="1">34.40 (2.10)</td><td valign="middle" align="center" rowspan="1" colspan="1">34.45 (3.36)</td><td valign="middle" align="center" rowspan="1" colspan="1">34.40 (2.10)</td></tr><tr><td valign="middle" align="right" rowspan="1" colspan="1">150</td><td valign="top" align="center" rowspan="1" colspan="1">32.68 (0.89)</td><td valign="top" align="center" rowspan="1" colspan="1">32.85 (1.19)</td><td valign="top" align="center" rowspan="1" colspan="1">32.92 (1.06)</td><td valign="top" align="center" rowspan="1" colspan="1">32.68 (0.89)</td><td valign="top" align="center" rowspan="1" colspan="1">32.82 (0.97)</td><td valign="top" align="center" rowspan="1" colspan="1">32.68 (0.89)</td></tr><tr><td valign="middle" align="right" rowspan="1" colspan="1">300</td><td valign="top" align="center" rowspan="1" colspan="1">31.91 (0.54)</td><td valign="top" align="center" rowspan="1" colspan="1">31.99 (0.88)</td><td valign="top" align="center" rowspan="1" colspan="1">31.97 (0.77)</td><td valign="top" align="center" rowspan="1" colspan="1">31.91 (0.54)</td><td valign="top" align="center" rowspan="1" colspan="1">31.98 (0.68)</td><td valign="top" align="center" rowspan="1" colspan="1">31.91 (0.54)</td></tr><tr><td valign="top" align="left" rowspan="3" colspan="1" style="background-color:rgb(238,242,249)">ORF1a</td><td valign="middle" align="right" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">50</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">33.93 (1.30)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">33.90 (0.86)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">34.07 (1.53)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">33.93 (1.30)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">34.14 (1.34)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">34.04 (1.21)</td></tr><tr><td valign="middle" align="right" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">150</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">32.16 (0.79)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">32.11 (0.63)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">32.23 (0.77)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">32.16 (0.79)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">32.31 (0.77)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">32.27 (0.49)</td></tr><tr><td valign="middle" align="right" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">300</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">30.88 (0.62)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">30.78 (0.62)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">30.91 (0.54)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">30.88 (0.62)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">30.85 (1.15)</td><td valign="top" align="center" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">30.98 (0.70)</td></tr></tbody></table></alternatives><table-wrap-foot><fn><p>Abbreviation: Ct, Cycle threshold.</p></fn></table-wrap-foot></table-wrap><table-wrap id="T4" orientation="portrait" position="float"><label>Table 4</label><caption><title>Comparison of the PowerChek MERS (upE and ORF1a) real-time PCR Kit and the sequencing assay for the detection of MERS-CoV</title></caption><alternatives><graphic xlink:href="alm-37-494-i004"></graphic><table frame="hsides" rules="rows"><col width="40%" span="1"></col><col width="10%" span="1"></col><col width="20%" span="1"></col><col width="20%" span="1"></col><col width="10%" span="1"></col><thead><tr><th valign="middle" align="left" rowspan="2" colspan="1" style="background-color:rgb(218,227,244)">Detection method</th><th valign="middle" align="center" rowspan="2" colspan="1" style="background-color:rgb(218,227,244)">Result</th><th valign="middle" align="center" rowspan="1" colspan="2" style="background-color:rgb(218,227,244)">Subject characteristics</th><th valign="middle" align="center" rowspan="2" colspan="1" style="background-color:rgb(218,227,244)">Total</th></tr><tr><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">Symptomatic<break></break>MERS-confirmed patients</th><th valign="middle" align="center" rowspan="1" colspan="1" style="background-color:rgb(218,227,244)">Asymptomatic<break></break>MERS-negative individuals</th></tr></thead><tbody><tr><td valign="top" align="left" rowspan="2" colspan="1">PowerChek MERS (upE and ORF1a) real-time PCR Kit</td><td valign="top" align="left" rowspan="1" colspan="1">Positive</td><td valign="top" align="right" rowspan="1" colspan="1">50</td><td valign="top" align="right" rowspan="1" colspan="1">0</td><td valign="top" align="right" rowspan="1" colspan="1">50</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1">Negative</td><td valign="top" align="right" rowspan="1" colspan="1">0</td><td valign="top" align="right" rowspan="1" colspan="1">50</td><td valign="top" align="right" rowspan="1" colspan="1">50</td></tr><tr><td valign="top" align="left" rowspan="2" colspan="1" style="background-color:rgb(238,242,249)">RdRp and N gene sequencing assay</td><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Positive</td><td valign="top" align="right" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">50</td><td valign="top" align="right" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">0</td><td valign="top" align="right" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">50</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">Negative</td><td valign="top" align="right" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">0</td><td valign="top" align="right" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">50</td><td valign="top" align="right" rowspan="1" colspan="1" style="background-color:rgb(238,242,249)">50</td></tr><tr><td valign="top" align="left" rowspan="1" colspan="1"></td><td valign="top" align="left" rowspan="1" colspan="1">Total</td><td valign="top" align="right" rowspan="1" colspan="1">50</td><td valign="top" align="right" rowspan="1" colspan="1">50</td><td valign="top" align="right" rowspan="1" colspan="1">100</td></tr></tbody></table></alternatives><table-wrap-foot><fn><p>Positive percent agreement, 100% (91.1–100%).</p><p>Negative percent agreement, 100% (91.1–100%).</p><p>Kappa coefficient, 1.00.</p><p>Abbreviations: MERS, Middle East respiratory syndrome; RdRp, RNA-dependent RNA polymerase.</p></fn></table-wrap-foot></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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