Analytical and Clinical Validation of Six Commercial Middle East Respiratory Syndrome Coronavirus RNA Detection Kits Based on Real-Time Reverse-Transcription PCR
Identifieur interne : 001376 ( Main/Exploration ); précédent : 001375; suivant : 001377Analytical and Clinical Validation of Six Commercial Middle East Respiratory Syndrome Coronavirus RNA Detection Kits Based on Real-Time Reverse-Transcription PCR
Auteurs : Mi-Na Kim [Corée du Sud] ; Young Jin Ko [Corée du Sud] ; Moon-Woo Seong [Corée du Sud] ; Jae-Seok Kim [Corée du Sud] ; Bo-Moon Shin [Corée du Sud] ; Heungsup Sung [Corée du Sud]Source :
- Annals of Laboratory Medicine [ 2234-3806 ] ; 2016.
Descripteurs français
- KwdFr :
- ARN viral (analyse), ARN viral (métabolisme), Cadres ouverts de lecture (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification), Humains, Infections à coronavirus (diagnostic), Infections à coronavirus (virologie), Partie nasale du pharynx (virologie), Protéines de l'enveloppe virale (génétique), Réaction de polymérisation en chaine en temps réel, 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, Protéines de l'enveloppe virale.
- isolement et purification : Coronavirus du syndrome respiratoire du Moyen-Orient.
- métabolisme : ARN viral.
- virologie : Infections à coronavirus, Partie nasale du pharynx.
- Humains, Réaction de polymérisation en chaine en temps réel, 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, Viral Envelope Proteins (genetics).
- MESH :
- chemical , analysis : RNA, Viral.
- diagnosis : Coronavirus Infections.
- genetics : Middle East Respiratory Syndrome Coronavirus, Open Reading Frames, Viral Envelope Proteins.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus.
- chemical , metabolism : RNA, Viral.
- virology : Coronavirus Infections, Nasopharynx.
- Humans, Reagent Kits, Diagnostic, Real-Time Polymerase Chain Reaction.
Abstract
During the 2015 outbreak of Middle East Respiratory Syndrome coronavirus (MERS-CoV), six different commercial MERS-CoV RNA detection kits based on real-time reverse-transcription polymerase chain reaction (rRT-PCR) were available in Korea. We performed analytical and clinical validations of these kits.
PowerChek (Kogene Biotech, Korea), DiaPlexQ (SolGent, Korea), Anyplex (Seegene, Korea), AccuPower (Bioneer, Korea), LightMix (Roche Molecular Diagnostics, Switzerland), and UltraFast kits (Nanobiosys, Korea) were evaluated. Limits of detection (LOD) with 95% probability values were estimated by testing 16 replicates of upstream of the envelope gene (
The LODs for
The overall sensitivity and specificity of all six assay systems were sufficient for diagnosing MERS-CoV infection. However, the analytical sensitivity and detection ability in specimens with PCR inhibition could be improved with the use of appropriate internal controls.
Url:
DOI: 10.3343/alm.2016.36.5.450
PubMed: 27374710
PubMed Central: 4940488
Affiliations:
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Le document en format XML
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Coronavirus RNA Detection Kits Based on Real-Time Reverse-Transcription PCR</title><author><name sortKey="Kim, Mi Na" sort="Kim, Mi Na" uniqKey="Kim M" first="Mi-Na" last="Kim">Mi-Na Kim</name><affiliation wicri:level="3"><nlm:aff id="A1">Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan 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="Ko, Young Jin" sort="Ko, Young Jin" uniqKey="Ko Y" first="Young Jin" last="Ko">Young Jin Ko</name><affiliation wicri:level="3"><nlm:aff id="A1">Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan 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="Seong, Moon Woo" sort="Seong, Moon Woo" uniqKey="Seong M" first="Moon-Woo" last="Seong">Moon-Woo Seong</name><affiliation wicri:level="3"><nlm:aff id="A2">Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul National University Hospital, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author><author><name sortKey="Kim, Jae Seok" sort="Kim, Jae Seok" uniqKey="Kim J" first="Jae-Seok" last="Kim">Jae-Seok Kim</name><affiliation wicri:level="3"><nlm:aff id="A3">Department of Laboratory Medicine, Hallym University College of Medicine, Hallym University Kangdong Sacred Heart Hospital, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine, Hallym University College of Medicine, Hallym University Kangdong Sacred Heart Hospital, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author><author><name sortKey="Shin, Bo Moon" sort="Shin, Bo Moon" uniqKey="Shin B" first="Bo-Moon" last="Shin">Bo-Moon Shin</name><affiliation wicri:level="3"><nlm:aff id="A4">Department of Laboratory Medicine, Inje University, Sanggye Paik Hospital, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine, Inje University, Sanggye Paik Hospital, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author><author><name sortKey="Sung, Heungsup" sort="Sung, Heungsup" uniqKey="Sung H" first="Heungsup" last="Sung">Heungsup Sung</name><affiliation wicri:level="3"><nlm:aff id="A1">Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Korea.</nlm:aff><country xml:lang="fr" wicri:curation="lc">Corée du Sud</country><wicri:regionArea>Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></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="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Coronavirus Infections (diagnosis)</term><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><term>Real-Time Polymerase Chain Reaction</term><term>Viral Envelope Proteins (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (analyse)</term><term>ARN viral (métabolisme)</term><term>Cadres ouverts de lecture (génétique)</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>Infections à coronavirus (virologie)</term><term>Partie nasale du pharynx (virologie)</term><term>Protéines de l'enveloppe virale (génétique)</term><term>Réaction de polymérisation en chaine en temps réel</term><term>Trousses de réactifs pour diagnostic</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ARN 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><term>Open Reading Frames</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Cadres ouverts de lecture</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term><term>Protéines de l'enveloppe virale</term></keywords><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><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN viral</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Infections à coronavirus</term><term>Partie nasale du pharynx</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Coronavirus Infections</term><term>Nasopharynx</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term><term>Reagent Kits, Diagnostic</term><term>Real-Time Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Humains</term><term>Réaction de polymérisation en chaine en temps réel</term><term>Trousses de réactifs pour diagnostic</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>During the 2015 outbreak of Middle East Respiratory Syndrome coronavirus (MERS-CoV), six different commercial MERS-CoV RNA detection kits based on real-time reverse-transcription polymerase chain reaction (rRT-PCR) were available in Korea. We performed analytical and clinical validations of these kits.</p></sec><sec><title>Methods</title><p>PowerChek (Kogene Biotech, Korea), DiaPlexQ (SolGent, Korea), Anyplex (Seegene, Korea), AccuPower (Bioneer, Korea), LightMix (Roche Molecular Diagnostics, Switzerland), and UltraFast kits (Nanobiosys, Korea) were evaluated. Limits of detection (LOD) with 95% probability values were estimated by testing 16 replicates of upstream of the envelope gene (<italic>upE</italic>) and open reading frame 1a (<italic>ORF1a</italic>) RNA transcripts. Specificity was estimated by using 28 nasopharyngeal swabs that were positive for other respiratory viruses. Clinical sensitivity was evaluated by using 18 lower respiratory specimens. The sensitivity test panel and the high inhibition panel were composed of nine specimens each, including eight and six specimens that were positive for MERS-CoV, respectively.</p></sec><sec><title>Results</title><p>The LODs for <italic>upE</italic> ranged from 21.88 to 263.03 copies/reaction, and those for <italic>ORF1a</italic> ranged from 6.92 to 128.82 copies/reaction. No cross-reactivity with other respiratory viruses was found. All six kits correctly identified 8 of 8 (100%) positive clinical specimens. Based on results from the high inhibition panel, PowerChek and AccuPower were the least sensitive to the presence of PCR inhibition.</p></sec><sec><title>Conclusions</title><p>The overall sensitivity and specificity of all six assay systems were sufficient for diagnosing MERS-CoV infection. However, the analytical sensitivity and detection ability in specimens with PCR inhibition could be improved with the use of appropriate internal controls.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Cowling, Bj" uniqKey="Cowling B">BJ Cowling</name></author><author><name sortKey="Park, M" uniqKey="Park M">M Park</name></author><author><name sortKey="Fang, Vj" uniqKey="Fang V">VJ Fang</name></author><author><name sortKey="Wu, P" uniqKey="Wu P">P Wu</name></author><author><name sortKey="Leung, Gm" uniqKey="Leung G">GM Leung</name></author><author><name sortKey="Wu, Jt" uniqKey="Wu J">JT Wu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Park, Hy" uniqKey="Park H">HY Park</name></author><author><name sortKey="Lee, Ej" uniqKey="Lee E">EJ Lee</name></author><author><name sortKey="Ryu, Yw" uniqKey="Ryu Y">YW Ryu</name></author><author><name sortKey="Kim, Y" uniqKey="Kim Y">Y Kim</name></author><author><name sortKey="Kim, H" uniqKey="Kim H">H Kim</name></author><author><name sortKey="Lee, H" uniqKey="Lee H">H Lee</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Lu, X" uniqKey="Lu X">X Lu</name></author><author><name sortKey="Whitaker, B" uniqKey="Whitaker B">B Whitaker</name></author><author><name sortKey="Sakthivel, Sk" uniqKey="Sakthivel S">SK Sakthivel</name></author><author><name sortKey="Kamili, S" uniqKey="Kamili S">S Kamili</name></author><author><name sortKey="Rose, Le" uniqKey="Rose L">LE Rose</name></author><author><name sortKey="Lowe, L" uniqKey="Lowe L">L Lowe</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Corman, Vm" uniqKey="Corman V">VM Corman</name></author><author><name sortKey="Muller, Ma" uniqKey="Muller M">MA Müller</name></author><author><name sortKey="Costabel, U" uniqKey="Costabel U">U Costabel</name></author><author><name sortKey="Timm, J" uniqKey="Timm J">J Timm</name></author><author><name sortKey="Binger, T" uniqKey="Binger T">T Binger</name></author><author><name sortKey="Meyer, B" uniqKey="Meyer B">B Meyer</name></author></analytic></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" uniqKey="Zaki A">A Zaki</name></author><author><name sortKey="Landt, O" uniqKey="Landt O">O Landt</name></author><author><name sortKey="Eschbach Bludau, M" uniqKey="Eschbach Bludau M">M Eschbach-Bludau</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Corman, Vm" uniqKey="Corman V">VM Corman</name></author><author><name sortKey="Olschl Ger, S" uniqKey="Olschl Ger S">S Ölschläger</name></author><author><name sortKey="Wendtner, Cm" uniqKey="Wendtner C">CM Wendtner</name></author><author><name sortKey="Drexler, Jf" uniqKey="Drexler J">JF Drexler</name></author><author><name sortKey="Hess, M" uniqKey="Hess M">M Hess</name></author><author><name sortKey="Drosten, C" uniqKey="Drosten C">C Drosten</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Madej, Rm" uniqKey="Madej R">RM Madej</name></author><author><name sortKey="Davis, J" uniqKey="Davis J">J Davis</name></author><author><name sortKey="Holden, Mj" uniqKey="Holden M">MJ Holden</name></author><author><name sortKey="Kwang, S" uniqKey="Kwang S">S Kwang</name></author><author><name sortKey="Labourier, E" uniqKey="Labourier E">E Labourier</name></author><author><name sortKey="Schneider, Gj" uniqKey="Schneider G">GJ Schneider</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kraft, Cs" uniqKey="Kraft C">CS Kraft</name></author><author><name sortKey="Armstrong, Ws" uniqKey="Armstrong W">WS Armstrong</name></author><author><name sortKey="Caliendo, Am" uniqKey="Caliendo A">AM Caliendo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Schnepf, N" uniqKey="Schnepf N">N Schnepf</name></author><author><name sortKey="Scieux, C" uniqKey="Scieux C">C Scieux</name></author><author><name sortKey="Resche Riggon, M" uniqKey="Resche Riggon M">M Resche-Riggon</name></author><author><name sortKey="Feghoul, L" uniqKey="Feghoul L">L Feghoul</name></author><author><name sortKey="Xhaard, A" uniqKey="Xhaard A">A Xhaard</name></author><author><name sortKey="Gallien, S" uniqKey="Gallien S">S Gallien</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cotten, M" uniqKey="Cotten M">M Cotten</name></author></analytic></biblStruct><biblStruct></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Corée du Sud</li></country><region><li>Région capitale de Séoul</li></region><settlement><li>Séoul</li></settlement></list><tree><country name="Corée du Sud"><region name="Région capitale de Séoul"><name sortKey="Kim, Mi Na" sort="Kim, Mi Na" uniqKey="Kim M" first="Mi-Na" last="Kim">Mi-Na Kim</name></region><name sortKey="Kim, Jae Seok" sort="Kim, Jae Seok" uniqKey="Kim J" first="Jae-Seok" last="Kim">Jae-Seok Kim</name><name sortKey="Ko, Young Jin" sort="Ko, Young Jin" uniqKey="Ko Y" first="Young Jin" last="Ko">Young Jin Ko</name><name sortKey="Seong, Moon Woo" sort="Seong, Moon Woo" uniqKey="Seong M" first="Moon-Woo" last="Seong">Moon-Woo Seong</name><name sortKey="Shin, Bo Moon" sort="Shin, Bo Moon" uniqKey="Shin B" first="Bo-Moon" last="Shin">Bo-Moon Shin</name><name sortKey="Sung, Heungsup" sort="Sung, Heungsup" uniqKey="Sung H" first="Heungsup" last="Sung">Heungsup Sung</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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