Development of a molecular-beacon-based multi-allelic real-time RT-PCR assay for the detection of human coronavirus causing severe acute respiratory syndrome (SARS-CoV): a general methodology for detecting rapidly mutating viruses
Identifieur interne : 002300 ( Main/Exploration ); précédent : 002299; suivant : 002301Development of a molecular-beacon-based multi-allelic real-time RT-PCR assay for the detection of human coronavirus causing severe acute respiratory syndrome (SARS-CoV): a general methodology for detecting rapidly mutating viruses
Auteurs : Andreas V. Hadjinicolaou [Chypre (pays)] ; Gabriella A. Farcas [Canada] ; Victoria L. Demetriou [Chypre (pays)] ; Tony Mazzulli [Canada] ; Susan M. Poutanen [Canada] ; Barbara M. Willey [Canada] ; Donald E. Low [Canada] ; Jagdish Butany [Canada] ; Sylvia L. Asa [Canada] ; Kevin C. Kain [Canada] ; Leondios G. Kostrikis [Chypre (pays)]Source :
- Archives of Virology [ 0304-8608 ] ; 2011-04-01.
Descripteurs français
- KwdFr :
- Humains, Mutation, RT-PCR (), Sensibilité et spécificité, Sondes oligonucléotidiques (), Sondes oligonucléotidiques (génétique), Syndrome respiratoire aigu sévère (diagnostic), Techniques de diagnostic moléculaire (), Virologie (), Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Sondes oligonucléotidiques, Virus du SRAS.
- isolement et purification : Virus du SRAS.
- Humains, Mutation, RT-PCR, Sensibilité et spécificité, Sondes oligonucléotidiques, Techniques de diagnostic moléculaire, Virologie.
- Pascal (Inist)
- Wicri :
- topic : Homme.
English descriptors
- KwdEn :
- Coronavirus, Detection, Human, Humans, Molecular Diagnostic Techniques (methods), Mutation, Oligonucleotide Probes (chemistry), Oligonucleotide Probes (genetics), Real time, Reverse Transcriptase Polymerase Chain Reaction (methods), Reverse transcription polymerase chain reaction, SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe acute respiratory syndrome, Severe acute respiratory syndrome virus, Virology (methods).
- MESH :
- chemical , chemistry : Oligonucleotide Probes.
- chemical , genetics : Oligonucleotide Probes.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Molecular Diagnostic Techniques, Reverse Transcriptase Polymerase Chain Reaction, Virology.
- Humans, Mutation, Sensitivity and Specificity.
Abstract
Abstract: Emerging infectious diseases have caused a global effort for development of fast and accurate detection techniques. The rapidly mutating nature of viruses presents a major difficulty, highlighting the need for specific detection of genetically diverse strains. One such infectious agent is SARS-associated coronavirus (SARS-CoV), which emerged in 2003. This study aimed to develop a real-time RT-PCR detection assay specific for SARS-CoV, taking into account its intrinsic polymorphic nature due to genetic drift and recombination and the possibility of continuous and multiple introductions of genetically non-identical strains into the human population, by using mismatch-tolerant molecular beacons designed to specifically detect the SARS-CoV S, E, M and N genes. These were applied in simple, reproducible duplex and multiplex real-time PCR assays on 25 post-mortem samples and constructed RNA controls, and they demonstrated high target detection ability and specificity. This assay can readily be adapted for detection of other emerging and rapidly mutating pathogens.
Url:
- https://api.istex.fr/ark:/67375/VQC-2DM8TFP6-C/fulltext.pdf
- http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7087183
DOI: 10.1007/s00705-010-0906-7
Affiliations:
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Le document en format XML
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Asa</name><affiliation wicri:level="4"><country xml:lang="fr">Canada</country><wicri:regionArea>University of Toronto, Toronto, ON</wicri:regionArea><orgName type="university">Université de Toronto</orgName><placeName><settlement type="city">Toronto</settlement><region type="state">Ontario</region></placeName></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Canada</country><wicri:regionArea>Toronto Medical Laboratories, Toronto, ON</wicri:regionArea><wicri:noRegion>ON</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Canada</country><wicri:regionArea>University Health Network, Toronto, ON</wicri:regionArea><wicri:noRegion>ON</wicri:noRegion></affiliation></author><author><name sortKey="Kain, Kevin C" sort="Kain, Kevin C" uniqKey="Kain K" first="Kevin C." last="Kain">Kevin C. Kain</name><affiliation wicri:level="4"><country xml:lang="fr">Canada</country><wicri:regionArea>University of Toronto, Toronto, ON</wicri:regionArea><orgName type="university">Université de Toronto</orgName><placeName><settlement type="city">Toronto</settlement><region type="state">Ontario</region></placeName></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Canada</country><wicri:regionArea>Toronto General Hospital, Toronto, ON</wicri:regionArea><wicri:noRegion>ON</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Canada</country><wicri:regionArea>Mount Sinai Hospital, Toronto, ON</wicri:regionArea><wicri:noRegion>ON</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Canada</country><wicri:regionArea>University Health Network, Toronto, ON</wicri:regionArea><wicri:noRegion>ON</wicri:noRegion></affiliation></author><author><name sortKey="Kostrikis, Leondios G" sort="Kostrikis, Leondios G" uniqKey="Kostrikis L" first="Leondios G." last="Kostrikis">Leondios G. Kostrikis</name><affiliation wicri:level="1"><country xml:lang="fr">Chypre (pays)</country><wicri:regionArea>Laboratory of Biotechnology and Molecular Virology, Department of Biological Sciences, University of Cyprus, 75 Kallipoleos Avenue, PO Box 20537, 1678, Nicosia</wicri:regionArea><wicri:noRegion>Nicosia</wicri:noRegion></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Chypre (pays)</country></affiliation></author></analytic><monogr></monogr><series><title level="j">Archives of Virology</title><title level="j" type="sub">Official Journal of the Virology Division of the International Union of Microbiological Societies</title><title level="j" type="abbrev">Arch Virol</title><idno type="ISSN">0304-8608</idno><idno type="eISSN">1432-8798</idno><imprint><publisher>Springer Vienna</publisher><pubPlace>Vienna</pubPlace><date type="published" when="2011-04-01">2011-04-01</date><biblScope unit="volume">156</biblScope><biblScope unit="issue">4</biblScope><biblScope unit="page" from="671">671</biblScope><biblScope unit="page" to="680">680</biblScope></imprint><idno type="ISSN">0304-8608</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0304-8608</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Coronavirus</term><term>Detection</term><term>Human</term><term>Humans</term><term>Molecular Diagnostic Techniques (methods)</term><term>Mutation</term><term>Oligonucleotide Probes (chemistry)</term><term>Oligonucleotide Probes (genetics)</term><term>Real time</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term><term>Reverse transcription polymerase chain reaction</term><term>SARS Virus (genetics)</term><term>SARS Virus (isolation & purification)</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe acute respiratory syndrome</term><term>Severe acute respiratory syndrome virus</term><term>Virology (methods)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Humains</term><term>Mutation</term><term>RT-PCR ()</term><term>Sensibilité et spécificité</term><term>Sondes oligonucléotidiques ()</term><term>Sondes oligonucléotidiques (génétique)</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Techniques de diagnostic moléculaire ()</term><term>Virologie ()</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Oligonucleotide Probes</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Oligonucleotide Probes</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Sondes oligonucléotidiques</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Molecular Diagnostic Techniques</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Virology</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term><term>Mutation</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Humains</term><term>Mutation</term><term>RT-PCR</term><term>Sensibilité et spécificité</term><term>Sondes oligonucléotidiques</term><term>Techniques de diagnostic moléculaire</term><term>Virologie</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term><term>Détection</term><term>Homme</term><term>Réaction chaîne polymérase RT</term><term>Syndrome respiratoire aigu sévère</term><term>Temps réel</term><term>Virus syndrome respiratoire aigu sévère</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Homme</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Emerging infectious diseases have caused a global effort for development of fast and accurate detection techniques. The rapidly mutating nature of viruses presents a major difficulty, highlighting the need for specific detection of genetically diverse strains. One such infectious agent is SARS-associated coronavirus (SARS-CoV), which emerged in 2003. This study aimed to develop a real-time RT-PCR detection assay specific for SARS-CoV, taking into account its intrinsic polymorphic nature due to genetic drift and recombination and the possibility of continuous and multiple introductions of genetically non-identical strains into the human population, by using mismatch-tolerant molecular beacons designed to specifically detect the SARS-CoV S, E, M and N genes. These were applied in simple, reproducible duplex and multiplex real-time PCR assays on 25 post-mortem samples and constructed RNA controls, and they demonstrated high target detection ability and specificity. This assay can readily be adapted for detection of other emerging and rapidly mutating pathogens.</div></front></TEI><affiliations><list><country><li>Canada</li><li>Chypre (pays)</li></country><region><li>Ontario</li></region><settlement><li>Toronto</li></settlement><orgName><li>Université de Toronto</li></orgName></list><tree><country name="Chypre (pays)"><noRegion><name sortKey="Hadjinicolaou, Andreas V" sort="Hadjinicolaou, Andreas V" uniqKey="Hadjinicolaou A" first="Andreas V." last="Hadjinicolaou">Andreas V. Hadjinicolaou</name></noRegion><name sortKey="Demetriou, Victoria L" sort="Demetriou, Victoria L" uniqKey="Demetriou V" first="Victoria L." last="Demetriou">Victoria L. Demetriou</name><name sortKey="Kostrikis, Leondios G" sort="Kostrikis, Leondios G" uniqKey="Kostrikis L" first="Leondios G." last="Kostrikis">Leondios G. Kostrikis</name><name sortKey="Kostrikis, Leondios G" sort="Kostrikis, Leondios G" uniqKey="Kostrikis L" first="Leondios G." last="Kostrikis">Leondios G. Kostrikis</name></country><country name="Canada"><region name="Ontario"><name sortKey="Farcas, Gabriella A" sort="Farcas, Gabriella A" uniqKey="Farcas G" first="Gabriella A." last="Farcas">Gabriella A. Farcas</name></region><name sortKey="Asa, Sylvia L" sort="Asa, Sylvia L" uniqKey="Asa S" first="Sylvia L." last="Asa">Sylvia L. Asa</name><name sortKey="Asa, Sylvia L" sort="Asa, Sylvia L" uniqKey="Asa S" first="Sylvia L." last="Asa">Sylvia L. Asa</name><name sortKey="Asa, Sylvia L" sort="Asa, Sylvia L" uniqKey="Asa S" first="Sylvia L." last="Asa">Sylvia L. Asa</name><name sortKey="Butany, Jagdish" sort="Butany, Jagdish" uniqKey="Butany J" first="Jagdish" last="Butany">Jagdish Butany</name><name sortKey="Butany, Jagdish" sort="Butany, Jagdish" uniqKey="Butany J" first="Jagdish" last="Butany">Jagdish Butany</name><name sortKey="Butany, Jagdish" sort="Butany, Jagdish" uniqKey="Butany J" first="Jagdish" last="Butany">Jagdish Butany</name><name sortKey="Farcas, Gabriella A" sort="Farcas, Gabriella A" uniqKey="Farcas G" first="Gabriella A." last="Farcas">Gabriella A. Farcas</name><name sortKey="Kain, Kevin C" sort="Kain, Kevin C" uniqKey="Kain K" first="Kevin C." last="Kain">Kevin C. Kain</name><name sortKey="Kain, Kevin C" sort="Kain, Kevin C" uniqKey="Kain K" first="Kevin C." last="Kain">Kevin C. Kain</name><name sortKey="Kain, Kevin C" sort="Kain, Kevin C" uniqKey="Kain K" first="Kevin C." last="Kain">Kevin C. Kain</name><name sortKey="Kain, Kevin C" sort="Kain, Kevin C" uniqKey="Kain K" first="Kevin C." last="Kain">Kevin C. Kain</name><name sortKey="Low, Donald E" sort="Low, Donald E" uniqKey="Low D" first="Donald E." last="Low">Donald E. Low</name><name sortKey="Low, Donald E" sort="Low, Donald E" uniqKey="Low D" first="Donald E." last="Low">Donald E. Low</name><name sortKey="Low, Donald E" sort="Low, Donald E" uniqKey="Low D" first="Donald E." last="Low">Donald E. Low</name><name sortKey="Mazzulli, Tony" sort="Mazzulli, Tony" uniqKey="Mazzulli T" first="Tony" last="Mazzulli">Tony Mazzulli</name><name sortKey="Mazzulli, Tony" sort="Mazzulli, Tony" uniqKey="Mazzulli T" first="Tony" last="Mazzulli">Tony Mazzulli</name><name sortKey="Mazzulli, Tony" sort="Mazzulli, Tony" uniqKey="Mazzulli T" first="Tony" last="Mazzulli">Tony Mazzulli</name><name sortKey="Poutanen, Susan M" sort="Poutanen, Susan M" uniqKey="Poutanen S" first="Susan M." last="Poutanen">Susan M. Poutanen</name><name sortKey="Poutanen, Susan M" sort="Poutanen, Susan M" uniqKey="Poutanen S" first="Susan M." last="Poutanen">Susan M. Poutanen</name><name sortKey="Poutanen, Susan M" sort="Poutanen, Susan M" uniqKey="Poutanen S" first="Susan M." last="Poutanen">Susan M. Poutanen</name><name sortKey="Willey, Barbara M" sort="Willey, Barbara M" uniqKey="Willey B" first="Barbara M." last="Willey">Barbara M. Willey</name><name sortKey="Willey, Barbara M" sort="Willey, Barbara M" uniqKey="Willey B" first="Barbara M." last="Willey">Barbara M. Willey</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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