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 : 002277 ( Ncbi/Curation ); précédent : 002276; suivant : 002278Development 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 ; Victoria L. Demetriou [Chypre (pays)] ; Tony Mazzulli ; Susan M. Poutanen ; Barbara M. Willey ; Donald E. Low ; Jagdish Butany ; Sylvia L. Asa ; Kevin C. Kain ; Leondios G. Kostrikis [Chypre (pays)]Source :
- Archives of Virology [ 0304-8608 ] ; 2011.
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.
English descriptors
- KwdEn :
- Humans, Molecular Diagnostic Techniques (methods), Mutation, Oligonucleotide Probes (chemistry), Oligonucleotide Probes (genetics), Reverse Transcriptase Polymerase Chain Reaction (methods), SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), 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
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
Url:
DOI: 10.1007/s00705-010-0906-7
PubMed: 21221674
PubMed Central: 7087183
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Gabriella A. 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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">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</title><author><name sortKey="Hadjinicolaou, Andreas V" sort="Hadjinicolaou, Andreas V" uniqKey="Hadjinicolaou A" first="Andreas V." last="Hadjinicolaou">Andreas V. Hadjinicolaou</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.6603.3</institution-id><institution-id institution-id-type="ISNI">0000000121167908</institution-id><institution>Laboratory of Biotechnology and Molecular Virology, Department of Biological Sciences,</institution><institution>University of Cyprus,</institution></institution-wrap>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia, Cyprus</nlm:aff><country xml:lang="fr">Chypre (pays)</country><wicri:regionArea>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia</wicri:regionArea><wicri:noRegion>1678 Nicosia</wicri:noRegion></affiliation></author><author><name sortKey="Farcas, Gabriella A" sort="Farcas, Gabriella A" uniqKey="Farcas G" first="Gabriella A." last="Farcas">Gabriella A. Farcas</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="GRID">grid.417184.f</institution-id><institution-id institution-id-type="ISNI">0000000106611177</institution-id><institution>Toronto General Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Demetriou, Victoria L" sort="Demetriou, Victoria L" uniqKey="Demetriou V" first="Victoria L." last="Demetriou">Victoria L. Demetriou</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.6603.3</institution-id><institution-id institution-id-type="ISNI">0000000121167908</institution-id><institution>Laboratory of Biotechnology and Molecular Virology, Department of Biological Sciences,</institution><institution>University of Cyprus,</institution></institution-wrap>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia, Cyprus</nlm:aff><country xml:lang="fr">Chypre (pays)</country><wicri:regionArea>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia</wicri:regionArea><wicri:noRegion>1678 Nicosia</wicri:noRegion></affiliation></author><author><name sortKey="Mazzulli, Tony" sort="Mazzulli, Tony" uniqKey="Mazzulli T" first="Tony" last="Mazzulli">Tony Mazzulli</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Poutanen, Susan M" sort="Poutanen, Susan M" uniqKey="Poutanen S" first="Susan M." last="Poutanen">Susan M. Poutanen</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Willey, Barbara M" sort="Willey, Barbara M" uniqKey="Willey B" first="Barbara M." last="Willey">Barbara M. Willey</name><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Low, Donald E" sort="Low, Donald E" uniqKey="Low D" first="Donald E." last="Low">Donald E. Low</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Butany, Jagdish" sort="Butany, Jagdish" uniqKey="Butany J" first="Jagdish" last="Butany">Jagdish Butany</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff6"><institution-wrap><institution-id institution-id-type="GRID">grid.231844.8</institution-id><institution-id institution-id-type="ISNI">0000000404740428</institution-id><institution>University Health Network,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Asa, Sylvia L" sort="Asa, Sylvia L" uniqKey="Asa S" first="Sylvia L." last="Asa">Sylvia L. Asa</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff6"><institution-wrap><institution-id institution-id-type="GRID">grid.231844.8</institution-id><institution-id institution-id-type="ISNI">0000000404740428</institution-id><institution>University Health Network,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></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><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="GRID">grid.417184.f</institution-id><institution-id institution-id-type="ISNI">0000000106611177</institution-id><institution>Toronto General Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff6"><institution-wrap><institution-id institution-id-type="GRID">grid.231844.8</institution-id><institution-id institution-id-type="ISNI">0000000404740428</institution-id><institution>University Health Network,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></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"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.6603.3</institution-id><institution-id institution-id-type="ISNI">0000000121167908</institution-id><institution>Laboratory of Biotechnology and Molecular Virology, Department of Biological Sciences,</institution><institution>University of Cyprus,</institution></institution-wrap>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia, Cyprus</nlm:aff><country xml:lang="fr">Chypre (pays)</country><wicri:regionArea>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia</wicri:regionArea><wicri:noRegion>1678 Nicosia</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">21221674</idno><idno type="pmc">7087183</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7087183</idno><idno type="RBID">PMC:7087183</idno><idno type="doi">10.1007/s00705-010-0906-7</idno><date when="2011">2011</date><idno type="wicri:Area/Pmc/Corpus">000267</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000267</idno><idno type="wicri:Area/Pmc/Curation">000267</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000267</idno><idno type="wicri:Area/Pmc/Checkpoint">000B52</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000B52</idno><idno type="wicri:source">PubMed</idno><idno type="RBID">pubmed:21221674</idno><idno type="wicri:Area/PubMed/Corpus">001565</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001565</idno><idno type="wicri:Area/PubMed/Curation">001565</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001565</idno><idno type="wicri:Area/PubMed/Checkpoint">001510</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001510</idno><idno type="wicri:Area/Ncbi/Merge">002277</idno><idno type="wicri:Area/Ncbi/Curation">002277</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">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</title><author><name sortKey="Hadjinicolaou, Andreas V" sort="Hadjinicolaou, Andreas V" uniqKey="Hadjinicolaou A" first="Andreas V." last="Hadjinicolaou">Andreas V. Hadjinicolaou</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.6603.3</institution-id><institution-id institution-id-type="ISNI">0000000121167908</institution-id><institution>Laboratory of Biotechnology and Molecular Virology, Department of Biological Sciences,</institution><institution>University of Cyprus,</institution></institution-wrap>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia, Cyprus</nlm:aff><country xml:lang="fr">Chypre (pays)</country><wicri:regionArea>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia</wicri:regionArea><wicri:noRegion>1678 Nicosia</wicri:noRegion></affiliation></author><author><name sortKey="Farcas, Gabriella A" sort="Farcas, Gabriella A" uniqKey="Farcas G" first="Gabriella A." last="Farcas">Gabriella A. Farcas</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="GRID">grid.417184.f</institution-id><institution-id institution-id-type="ISNI">0000000106611177</institution-id><institution>Toronto General Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Demetriou, Victoria L" sort="Demetriou, Victoria L" uniqKey="Demetriou V" first="Victoria L." last="Demetriou">Victoria L. Demetriou</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.6603.3</institution-id><institution-id institution-id-type="ISNI">0000000121167908</institution-id><institution>Laboratory of Biotechnology and Molecular Virology, Department of Biological Sciences,</institution><institution>University of Cyprus,</institution></institution-wrap>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia, Cyprus</nlm:aff><country xml:lang="fr">Chypre (pays)</country><wicri:regionArea>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia</wicri:regionArea><wicri:noRegion>1678 Nicosia</wicri:noRegion></affiliation></author><author><name sortKey="Mazzulli, Tony" sort="Mazzulli, Tony" uniqKey="Mazzulli T" first="Tony" last="Mazzulli">Tony Mazzulli</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Poutanen, Susan M" sort="Poutanen, Susan M" uniqKey="Poutanen S" first="Susan M." last="Poutanen">Susan M. Poutanen</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Willey, Barbara M" sort="Willey, Barbara M" uniqKey="Willey B" first="Barbara M." last="Willey">Barbara M. Willey</name><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Low, Donald E" sort="Low, Donald E" uniqKey="Low D" first="Donald E." last="Low">Donald E. Low</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Butany, Jagdish" sort="Butany, Jagdish" uniqKey="Butany J" first="Jagdish" last="Butany">Jagdish Butany</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff6"><institution-wrap><institution-id institution-id-type="GRID">grid.231844.8</institution-id><institution-id institution-id-type="ISNI">0000000404740428</institution-id><institution>University Health Network,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation></author><author><name sortKey="Asa, Sylvia L" sort="Asa, Sylvia L" uniqKey="Asa S" first="Sylvia L." last="Asa">Sylvia L. Asa</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff5">Toronto Medical Laboratories, Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff6"><institution-wrap><institution-id institution-id-type="GRID">grid.231844.8</institution-id><institution-id institution-id-type="ISNI">0000000404740428</institution-id><institution>University Health Network,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></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><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.17063.33</institution-id><institution-id institution-id-type="ISNI">0000 0001 2157 2938</institution-id><institution>University of Toronto,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="GRID">grid.417184.f</institution-id><institution-id institution-id-type="ISNI">0000000106611177</institution-id><institution>Toronto General Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.416166.2</institution-id><institution-id institution-id-type="ISNI">0000000404739881</institution-id><institution>Mount Sinai Hospital,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></affiliation><affiliation><nlm:aff id="Aff6"><institution-wrap><institution-id institution-id-type="GRID">grid.231844.8</institution-id><institution-id institution-id-type="ISNI">0000000404740428</institution-id><institution>University Health Network,</institution></institution-wrap>Toronto, ON Canada</nlm:aff><wicri:noCountry code="subfield">ON Canada</wicri:noCountry></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"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.6603.3</institution-id><institution-id institution-id-type="ISNI">0000000121167908</institution-id><institution>Laboratory of Biotechnology and Molecular Virology, Department of Biological Sciences,</institution><institution>University of Cyprus,</institution></institution-wrap>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia, Cyprus</nlm:aff><country xml:lang="fr">Chypre (pays)</country><wicri:regionArea>75 Kallipoleos Avenue, PO Box 20537, 1678 Nicosia</wicri:regionArea><wicri:noRegion>1678 Nicosia</wicri:noRegion></affiliation></author></analytic><series><title level="j">Archives of Virology</title><idno type="ISSN">0304-8608</idno><idno type="eISSN">1432-8798</idno><imprint><date when="2011">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Humans</term><term>Molecular Diagnostic Techniques (methods)</term><term>Mutation</term><term>Oligonucleotide Probes (chemistry)</term><term>Oligonucleotide Probes (genetics)</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</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>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></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>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 <italic>S, E, M</italic> and <italic>N</italic> 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.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Bressler, Am" uniqKey="Bressler A">AM Bressler</name></author><author><name sortKey="Nolte, Fs" uniqKey="Nolte F">FS Nolte</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Drosten, C" uniqKey="Drosten C">C Drosten</name></author><author><name sortKey="Gunther, S" uniqKey="Gunther S">S Gunther</name></author><author><name sortKey="Preiser, W" uniqKey="Preiser W">W Preiser</name></author><author><name sortKey="Van Der Werf, S" uniqKey="Van Der Werf S">S van der Werf</name></author><author><name sortKey="Brodt, Hr" uniqKey="Brodt H">HR Brodt</name></author><author><name sortKey="Becker, S" uniqKey="Becker S">S Becker</name></author><author><name sortKey="Rabenau, H" uniqKey="Rabenau H">H Rabenau</name></author><author><name sortKey="Panning, M" uniqKey="Panning M">M Panning</name></author><author><name sortKey="Kolesnikova, L" uniqKey="Kolesnikova L">L Kolesnikova</name></author><author><name sortKey="Fouchier, Ra" uniqKey="Fouchier R">RA Fouchier</name></author><author><name sortKey="Berger, A" uniqKey="Berger A">A Berger</name></author><author><name sortKey="Burguiere, Am" uniqKey="Burguiere A">AM Burguiere</name></author><author><name sortKey="Cinatl, J" uniqKey="Cinatl J">J Cinatl</name></author><author><name sortKey="Eickmann, M" uniqKey="Eickmann M">M Eickmann</name></author><author><name sortKey="Escriou, N" uniqKey="Escriou N">N Escriou</name></author><author><name sortKey="Grywna, K" uniqKey="Grywna K">K Grywna</name></author><author><name sortKey="Kramme, S" uniqKey="Kramme S">S Kramme</name></author><author><name sortKey="Manuguerra, Jc" uniqKey="Manuguerra J">JC Manuguerra</name></author><author><name sortKey="Muller, S" uniqKey="Muller S">S Muller</name></author><author><name sortKey="Rickerts, V" uniqKey="Rickerts V">V Rickerts</name></author><author><name sortKey="Sturmer, M" uniqKey="Sturmer M">M Sturmer</name></author><author><name sortKey="Vieth, S" uniqKey="Vieth S">S Vieth</name></author><author><name sortKey="Klenk, Hd" uniqKey="Klenk H">HD Klenk</name></author><author><name sortKey="Osterhaus, Ad" uniqKey="Osterhaus A">AD Osterhaus</name></author><author><name sortKey="Schmitz, H" uniqKey="Schmitz H">H Schmitz</name></author><author><name sortKey="Doerr, Hw" uniqKey="Doerr H">HW Doerr</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="El Hajj, Hh" uniqKey="El Hajj H">HH El-Hajj</name></author><author><name sortKey="Marras, Sa" uniqKey="Marras S">SA Marras</name></author><author><name sortKey="Tyagi, S" uniqKey="Tyagi S">S Tyagi</name></author><author><name sortKey="Shashkina, E" uniqKey="Shashkina E">E Shashkina</name></author><author><name sortKey="Kamboj, M" uniqKey="Kamboj M">M Kamboj</name></author><author><name sortKey="Kiehn, Te" 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sortKey="Zhang, S" uniqKey="Zhang S">S Zhang</name></author><author><name sortKey="Field, H" uniqKey="Field H">H Field</name></author><author><name sortKey="Daszak, P" uniqKey="Daszak P">P Daszak</name></author><author><name sortKey="Eaton, Bt" uniqKey="Eaton B">BT Eaton</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Weidmann, M" uniqKey="Weidmann M">M Weidmann</name></author><author><name sortKey="Zanotto, Pm" uniqKey="Zanotto P">PM Zanotto</name></author><author><name sortKey="Weber, F" uniqKey="Weber F">F Weber</name></author><author><name sortKey="Spiegel, M" uniqKey="Spiegel M">M Spiegel</name></author><author><name sortKey="Brodt, Hr" uniqKey="Brodt H">HR Brodt</name></author><author><name sortKey="Hufert, Ft" uniqKey="Hufert F">FT Hufert</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wu, Q" uniqKey="Wu Q">Q Wu</name></author><author><name sortKey="Xu, Z" uniqKey="Xu Z">Z Xu</name></author><author><name sortKey="Wei, T" uniqKey="Wei T">T Wei</name></author><author><name sortKey="Zeng, H" uniqKey="Zeng H">H Zeng</name></author><author><name sortKey="Li, J" uniqKey="Li J">J Li</name></author><author><name sortKey="Gang, H" uniqKey="Gang H">H Gang</name></author><author><name sortKey="Sun, M" uniqKey="Sun M">M Sun</name></author><author><name sortKey="Jiang, F" uniqKey="Jiang F">F Jiang</name></author><author><name sortKey="Wang, X" uniqKey="Wang X">X Wang</name></author><author><name sortKey="Dong, W" uniqKey="Dong W">W Dong</name></author><author><name sortKey="Yang, L" uniqKey="Yang L">L Yang</name></author><author><name sortKey="Wang, J" uniqKey="Wang J">J Wang</name></author></analytic></biblStruct></listBibl></div1></back></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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