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.
Url:
DOI: 10.3343/alm.2017.37.6.494
PubMed: 28840986
PubMed Central: 5587821
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<series><title level="j">Annals of Laboratory Medicine</title>
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<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>
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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>
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<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>
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<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>
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<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>
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