Performance Evaluation of the PowerChek MERS (upE & ORF1a) Real-Time PCR Kit for the Detection of Middle East Respiratory Syndrome Coronavirus RNA
Identifieur interne : 001B50 ( Ncbi/Merge ); précédent : 001B49; suivant : 001B51Performance 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.
Descripteurs français
- KwdFr :
- ARN viral (analyse), ARN viral (métabolisme), Analyse de séquence d'ARN, Cadres ouverts de lecture (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification), Expectoration (virologie), Humains, Infections à coronavirus (diagnostic), Infections à coronavirus (virologie), Partie nasale du pharynx (virologie), Réaction de polymérisation en chaine en temps réel, Sensibilité et spécificité, Trousses de réactifs pour diagnostic.
- MESH :
- analyse : ARN viral.
- diagnostic : Infections à coronavirus.
- génétique : Cadres ouverts de lecture, Coronavirus du syndrome respiratoire du Moyen-Orient.
- isolement et purification : Coronavirus du syndrome respiratoire du Moyen-Orient.
- métabolisme : ARN viral.
- virologie : Expectoration, Infections à coronavirus, Partie nasale du pharynx.
- Analyse de séquence d'ARN, Humains, Réaction de polymérisation en chaine en temps réel, Sensibilité et spécificité, Trousses de réactifs pour diagnostic.
English descriptors
- KwdEn :
- Coronavirus Infections (diagnosis), Coronavirus Infections (virology), Humans, Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (isolation & purification), Nasopharynx (virology), Open Reading Frames (genetics), RNA, Viral (analysis), RNA, Viral (metabolism), Reagent Kits, Diagnostic, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, Sequence Analysis, RNA, Sputum (virology).
- MESH :
- chemical , analysis : RNA, Viral.
- diagnosis : Coronavirus Infections.
- genetics : Middle East Respiratory Syndrome Coronavirus, Open Reading Frames.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus.
- chemical , metabolism : RNA, Viral.
- virology : Coronavirus Infections, Nasopharynx, Sputum.
- Humans, Reagent Kits, Diagnostic, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, Sequence Analysis, RNA.
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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PMC:5587821Le document en format XML
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<term>Coronavirus du syndrome respiratoire du Moyen-Orient (génétique)</term>
<term>Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification)</term>
<term>Expectoration (virologie)</term>
<term>Humains</term>
<term>Infections à coronavirus (diagnostic)</term>
<term>Infections à coronavirus (virologie)</term>
<term>Partie nasale du pharynx (virologie)</term>
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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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<author><name sortKey="To, Kk" uniqKey="To K">KK To</name>
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<author><name sortKey="Kim, Ji Youn" sort="Kim, Ji Youn" uniqKey="Kim J" first="Ji-Youn" last="Kim">Ji-Youn Kim</name>
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<author><name sortKey="Kwon, Hyeon Jeong" sort="Kwon, Hyeon Jeong" uniqKey="Kwon H" first="Hyeon Jeong" last="Kwon">Hyeon Jeong Kwon</name>
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</author>
<author><name sortKey="Yun, Sun Ae" sort="Yun, Sun Ae" uniqKey="Yun S" first="Sun Ae" last="Yun">Sun Ae Yun</name>
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</placeName>
</affiliation>
</author>
<author><name sortKey="Lee, Myoung Keun" sort="Lee, Myoung Keun" uniqKey="Lee M" first="Myoung-Keun" last="Lee">Myoung-Keun Lee</name>
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<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>
<placeName><settlement type="city">Séoul</settlement>
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</author>
<author><name sortKey="Ki, Chang Seok" sort="Ki, Chang Seok" uniqKey="Ki C" first="Chang-Seok" last="Ki">Chang-Seok Ki</name>
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<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>
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<author><name sortKey="Lee, Nam Yong" sort="Lee, Nam Yong" uniqKey="Lee N" first="Nam Yong" last="Lee">Nam Yong Lee</name>
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<author><name sortKey="Kim, Jong Won" sort="Kim, Jong Won" uniqKey="Kim J" first="Jong-Won" last="Kim">Jong-Won Kim</name>
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<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>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">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="3"><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>
<placeName><settlement type="city">Séoul</settlement>
<region type="capital">Région capitale de Séoul</region>
</placeName>
</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="3"><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>
<placeName><settlement type="city">Séoul</settlement>
<region type="capital">Région capitale de Séoul</region>
</placeName>
</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="3"><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>
<placeName><settlement type="city">Séoul</settlement>
<region type="capital">Région capitale de Séoul</region>
</placeName>
</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="3"><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>
<placeName><settlement type="city">Séoul</settlement>
<region type="capital">Région capitale de Séoul</region>
</placeName>
</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="3"><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>
<placeName><settlement type="city">Séoul</settlement>
<region type="capital">Région capitale de Séoul</region>
</placeName>
</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="3"><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>
<placeName><settlement type="city">Séoul</settlement>
<region type="capital">Région capitale de Séoul</region>
</placeName>
</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="3"><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>
<placeName><settlement type="city">Séoul</settlement>
<region type="capital">Région capitale de Séoul</region>
</placeName>
</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="3"><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>
<placeName><settlement type="city">Séoul</settlement>
<region type="capital">Région capitale de Séoul</region>
</placeName>
</affiliation>
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</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>
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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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<author><name sortKey="Huh, Hee Jae" sort="Huh, Hee Jae" uniqKey="Huh H" first="Hee Jae" last="Huh">Hee Jae Huh</name>
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<series><title level="j">Annals of laboratory medicine</title>
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<term>Coronavirus Infections (virology)</term>
<term>Humans</term>
<term>Middle East Respiratory Syndrome Coronavirus (genetics)</term>
<term>Middle East Respiratory Syndrome Coronavirus (isolation & purification)</term>
<term>Nasopharynx (virology)</term>
<term>Open Reading Frames (genetics)</term>
<term>RNA, Viral (analysis)</term>
<term>RNA, Viral (metabolism)</term>
<term>Reagent Kits, Diagnostic</term>
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<term>Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification)</term>
<term>Expectoration (virologie)</term>
<term>Humains</term>
<term>Infections à coronavirus (diagnostic)</term>
<term>Infections à coronavirus (virologie)</term>
<term>Partie nasale du pharynx (virologie)</term>
<term>Réaction de polymérisation en chaine en temps réel</term>
<term>Sensibilité et spécificité</term>
<term>Trousses de réactifs pour diagnostic</term>
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<term>Open Reading Frames</term>
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<term>Coronavirus du syndrome respiratoire du Moyen-Orient</term>
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<keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term>
</keywords>
<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term>
</keywords>
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<term>Nasopharynx</term>
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<keywords scheme="MESH" xml:lang="en"><term>Humans</term>
<term>Reagent Kits, Diagnostic</term>
<term>Real-Time Polymerase Chain Reaction</term>
<term>Sensitivity and Specificity</term>
<term>Sequence Analysis, RNA</term>
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<front><div type="abstract" xml:lang="en">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.</div>
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