Performance and clinical validation of the RealStar® MERS-CoV Kit for detection of Middle East respiratory syndrome coronavirus RNA
Identifieur interne : 001B23 ( Main/Exploration ); précédent : 001B22; suivant : 001B24Performance and clinical validation of the RealStar® MERS-CoV Kit for detection of Middle East respiratory syndrome coronavirus RNA
Auteurs : Victor Max Corman [Allemagne] ; Stephan Ölschl Ger [Allemagne] ; Clemens-Martin Wendtner [Allemagne] ; Jan Felix Drexler [Allemagne] ; Markus Hess [Allemagne] ; Christian Drosten [Allemagne]Source :
- Journal of Clinical Virology [ 1386-6532 ] ; 2014.
Descripteurs français
- KwdFr :
- ARN viral (génétique), ARN viral (isolement et purification), Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification), Humains, Infections à coronavirus (diagnostic), Normes de référence, Réaction de polymérisation en chaine en temps réel (), Réaction de polymérisation en chaine en temps réel (normes), Réactions croisées, Sensibilité et spécificité, Techniques de diagnostic moléculaire (), Techniques de diagnostic moléculaire (normes).
- MESH :
- diagnostic : Infections à coronavirus.
- génétique : ARN viral, Coronavirus du syndrome respiratoire du Moyen-Orient.
- isolement et purification : ARN viral, Coronavirus du syndrome respiratoire du Moyen-Orient.
- normes : Réaction de polymérisation en chaine en temps réel, Techniques de diagnostic moléculaire.
- Humains, Normes de référence, Réaction de polymérisation en chaine en temps réel, Réactions croisées, Sensibilité et spécificité, Techniques de diagnostic moléculaire.
English descriptors
- KwdEn :
- Coronavirus Infections (diagnosis), Cross Reactions, Humans, Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (isolation & purification), Molecular Diagnostic Techniques (methods), Molecular Diagnostic Techniques (standards), RNA, Viral (genetics), RNA, Viral (isolation & purification), Real-Time Polymerase Chain Reaction (methods), Real-Time Polymerase Chain Reaction (standards), Reference Standards, Sensitivity and Specificity.
- MESH :
- chemical , genetics : RNA, Viral.
- diagnosis : Coronavirus Infections.
- genetics : Middle East Respiratory Syndrome Coronavirus.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus, RNA, Viral.
- methods : Molecular Diagnostic Techniques, Real-Time Polymerase Chain Reaction.
- standards : Molecular Diagnostic Techniques, Real-Time Polymerase Chain Reaction.
- Cross Reactions, Humans, Reference Standards, Sensitivity and Specificity.
Abstract
A highly pathogenic human coronavirus causing respiratory disease emerged in the Middle East region in 2012. In-house molecular diagnostic methods for this virus termed
The aim of this study was to improve the availability of existing nucleic acid amplification-based diagnostic methods for MERS-CoV infections by providing a real-time RT-PCR kit, including an internal control and two target regions recommended by the World Health Organization (WHO). And to validate this kit (RealStar® MERS-CoV RT-PCR kit 1.0, Altona Diagnostics GmbH, Hamburg, Germany) using clinical samples of one MERS-CoV case from Munich and respiratory samples of patients with other respiratory diseases.
An internal amplification control was included into the RT-PCR assays targeting the genomic region upstream of the
The performance of both RT-PCR assays included in the kit is comparable to the in-house assays. They show high analytical sensitivity (upE: 5.3 copies/reaction; ORF1A: 9.3 copies/reaction), no cross-reactivity with other respiratory pathogens and detected MERS-CoV RNA in patient samples in almost the same manner as the in-house versions.
The kit is a valuable tool for assisting in the rapid diagnosis, patient management and epidemiology of suspected MERS-CoV cases.
Url:
DOI: 10.1016/j.jcv.2014.03.012
PubMed: 24726679
PubMed Central: 7106532
Affiliations:
- Allemagne
- Bavière, District de Cologne, District de Haute-Bavière, Hambourg, Rhénanie-du-Nord-Westphalie
- Bonn, Hambourg, Munich
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Le document en format XML
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Corman</name><affiliation wicri:level="3"><nlm:aff id="aff0005">Institute of Virology, University of Bonn Medical Centre, 53127 Bonn, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute of Virology, University of Bonn Medical Centre, 53127 Bonn</wicri:regionArea><placeName><region type="land" nuts="1">Rhénanie-du-Nord-Westphalie</region><region type="district" nuts="2">District de Cologne</region><settlement type="city">Bonn</settlement></placeName></affiliation></author><author><name sortKey="Olschl Ger, Stephan" sort="Olschl Ger, Stephan" uniqKey="Olschl Ger S" first="Stephan" last="Ölschl Ger">Stephan Ölschl Ger</name><affiliation wicri:level="1"><nlm:aff id="aff0010">Altona Diagnostics GmbH, Mörkenstrasse 12, 22767 Hamburg, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Altona Diagnostics GmbH, Mörkenstrasse 12, 22767 Hamburg</wicri:regionArea><wicri:noRegion>22767 Hamburg</wicri:noRegion><placeName><settlement 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C" first="Christian" last="Drosten">Christian Drosten</name><affiliation wicri:level="3"><nlm:aff id="aff0005">Institute of Virology, University of Bonn Medical Centre, 53127 Bonn, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute of Virology, University of Bonn Medical Centre, 53127 Bonn</wicri:regionArea><placeName><region type="land" nuts="1">Rhénanie-du-Nord-Westphalie</region><region type="district" nuts="2">District de Cologne</region><settlement type="city">Bonn</settlement></placeName></affiliation></author></analytic><series><title level="j">Journal of Clinical Virology</title><idno type="ISSN">1386-6532</idno><idno type="eISSN">1873-5967</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Coronavirus Infections (diagnosis)</term><term>Cross Reactions</term><term>Humans</term><term>Middle East Respiratory Syndrome Coronavirus (genetics)</term><term>Middle East Respiratory Syndrome Coronavirus (isolation & purification)</term><term>Molecular Diagnostic Techniques (methods)</term><term>Molecular Diagnostic Techniques (standards)</term><term>RNA, Viral (genetics)</term><term>RNA, Viral (isolation & purification)</term><term>Real-Time Polymerase Chain Reaction (methods)</term><term>Real-Time Polymerase Chain Reaction (standards)</term><term>Reference Standards</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (génétique)</term><term>ARN viral (isolement et purification)</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (génétique)</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification)</term><term>Humains</term><term>Infections à coronavirus (diagnostic)</term><term>Normes de référence</term><term>Réaction de polymérisation en chaine en temps réel ()</term><term>Réaction de polymérisation en chaine en temps réel (normes)</term><term>Réactions croisées</term><term>Sensibilité et spécificité</term><term>Techniques de diagnostic moléculaire ()</term><term>Techniques de diagnostic moléculaire (normes)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Coronavirus Infections</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Infections à coronavirus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN viral</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>ARN viral</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Molecular Diagnostic Techniques</term><term>Real-Time Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="normes" xml:lang="fr"><term>Réaction de polymérisation en chaine en temps réel</term><term>Techniques de diagnostic moléculaire</term></keywords><keywords scheme="MESH" qualifier="standards" xml:lang="en"><term>Molecular Diagnostic Techniques</term><term>Real-Time Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cross Reactions</term><term>Humans</term><term>Reference Standards</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Humains</term><term>Normes de référence</term><term>Réaction de polymérisation en chaine en temps réel</term><term>Réactions croisées</term><term>Sensibilité et spécificité</term><term>Techniques de diagnostic moléculaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>A highly pathogenic human coronavirus causing respiratory disease emerged in the Middle East region in 2012. In-house molecular diagnostic methods for this virus termed <italic>Middle East respiratory syndrome coronavirus</italic> (MERS-CoV) allowed sensitive MERS-CoV RNA detection in patient samples. Fast diagnosis is important to manage human cases and trace possible contacts.</p></sec><sec><title>Objectives</title><p>The aim of this study was to improve the availability of existing nucleic acid amplification-based diagnostic methods for MERS-CoV infections by providing a real-time RT-PCR kit, including an internal control and two target regions recommended by the World Health Organization (WHO). And to validate this kit (RealStar<sup>®</sup> MERS-CoV RT-PCR kit 1.0, Altona Diagnostics GmbH, Hamburg, Germany) using clinical samples of one MERS-CoV case from Munich and respiratory samples of patients with other respiratory diseases.</p></sec><sec><title>Study design</title><p>An internal amplification control was included into the RT-PCR assays targeting the genomic region upstream of the <italic>Envelope</italic> gene (upE) and within open reading frame (ORF) 1A. Based on these assays, a ready-to-use real-time RT-PCR kit featuring both the upE and ORF1A assays was developed, validated and compared to the established in-house versions.</p></sec><sec><title>Results</title><p>The performance of both RT-PCR assays included in the kit is comparable to the in-house assays. They show high analytical sensitivity (upE: 5.3 copies/reaction; ORF1A: 9.3 copies/reaction), no cross-reactivity with other respiratory pathogens and detected MERS-CoV RNA in patient samples in almost the same manner as the in-house versions.</p></sec><sec><title>Conclusion</title><p>The kit is a valuable tool for assisting in the rapid diagnosis, patient management and epidemiology of suspected MERS-CoV cases.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Bermingham, A" uniqKey="Bermingham A">A. Bermingham</name></author><author><name sortKey="Chand, M A" uniqKey="Chand M">M.A. Chand</name></author><author><name sortKey="Brown, C S" uniqKey="Brown C">C.S. Brown</name></author><author><name sortKey="Aarons, E" uniqKey="Aarons E">E. Aarons</name></author><author><name sortKey="Tong, C" uniqKey="Tong C">C. Tong</name></author><author><name sortKey="Langrish, C" uniqKey="Langrish C">C. 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Hassan</name></author><author><name sortKey="Memish, Z A" uniqKey="Memish Z">Z.A. Memish</name></author><author><name sortKey="Muth, D" uniqKey="Muth D">D. Muth</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gierer, S" uniqKey="Gierer S">S. Gierer</name></author><author><name sortKey="Hofmann Winkler, H" uniqKey="Hofmann Winkler H">H. Hofmann-Winkler</name></author><author><name sortKey="Albuali, W H" uniqKey="Albuali W">W.H. Albuali</name></author><author><name sortKey="Bertram, S" uniqKey="Bertram S">S. Bertram</name></author><author><name sortKey="Al Rubaish, A M" uniqKey="Al Rubaish A">A.M. Al-Rubaish</name></author><author><name sortKey="Yousef, A A" uniqKey="Yousef A">A.A. Yousef</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Khuri Bulos, N" uniqKey="Khuri Bulos N">N. Khuri-Bulos</name></author><author><name sortKey="Payne, D C" uniqKey="Payne D">D.C. Payne</name></author><author><name sortKey="Xiaoyan, L" uniqKey="Xiaoyan L">L. Xiaoyan</name></author><author><name sortKey="Erdman, D" uniqKey="Erdman D">D. Erdman</name></author><author><name sortKey="Wang, L" uniqKey="Wang L">L. Wang</name></author><author><name sortKey="Faouri, S" uniqKey="Faouri S">S. Faouri</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="De Sousa, R" uniqKey="De Sousa R">R. de Sousa</name></author><author><name sortKey="Reusken, C" uniqKey="Reusken C">C. Reusken</name></author><author><name sortKey="Koopmans, M" uniqKey="Koopmans M">M. Koopmans</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cotten, M" uniqKey="Cotten M">M. Cotten</name></author><author><name sortKey="Watson, S J" uniqKey="Watson S">S.J. Watson</name></author><author><name sortKey="Zumla, A I" uniqKey="Zumla A">A.I. Zumla</name></author><author><name sortKey="Makhdoom, H Q" uniqKey="Makhdoom H">H.Q. Makhdoom</name></author><author><name sortKey="Palser, A L" uniqKey="Palser A">A.L. Palser</name></author><author><name sortKey="Ong, S H" uniqKey="Ong S">S.H. Ong</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chu, D K W" uniqKey="Chu D">D.K.W. Chu</name></author><author><name sortKey="Poon, L L M" uniqKey="Poon L">L.L.M. Poon</name></author><author><name sortKey="Gomaa, M M" uniqKey="Gomaa M">M.M. Gomaa</name></author><author><name sortKey="Shehata, M M" uniqKey="Shehata M">M.M. Shehata</name></author><author><name sortKey="Perera, R A P M" uniqKey="Perera R">R.A.P.M. Perera</name></author><author><name sortKey="Zeid, D A" uniqKey="Zeid D">D.A. Zeid</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Allemagne</li></country><region><li>Bavière</li><li>District de Cologne</li><li>District de Haute-Bavière</li><li>Hambourg</li><li>Rhénanie-du-Nord-Westphalie</li></region><settlement><li>Bonn</li><li>Hambourg</li><li>Munich</li></settlement></list><tree><country name="Allemagne"><region name="Rhénanie-du-Nord-Westphalie"><name sortKey="Corman, Victor Max" sort="Corman, Victor Max" uniqKey="Corman V" first="Victor Max" last="Corman">Victor Max Corman</name></region><name sortKey="Drexler, Jan Felix" sort="Drexler, Jan Felix" uniqKey="Drexler J" first="Jan Felix" last="Drexler">Jan Felix Drexler</name><name sortKey="Drosten, Christian" sort="Drosten, Christian" uniqKey="Drosten C" first="Christian" last="Drosten">Christian Drosten</name><name sortKey="Hess, Markus" sort="Hess, Markus" uniqKey="Hess M" first="Markus" last="Hess">Markus Hess</name><name sortKey="Olschl Ger, Stephan" sort="Olschl Ger, Stephan" uniqKey="Olschl Ger S" first="Stephan" last="Ölschl Ger">Stephan Ölschl Ger</name><name sortKey="Wendtner, Clemens Martin" sort="Wendtner, Clemens Martin" uniqKey="Wendtner C" first="Clemens-Martin" last="Wendtner">Clemens-Martin Wendtner</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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