Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR
Identifieur interne : 001064 ( Ncbi/Merge ); précédent : 001063; suivant : 001065Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR
Auteurs : Els Keyaerts [Belgique] ; Leen Vijgen [Belgique] ; Piet Maes [Belgique] ; Griet Duson [Belgique] ; Johan Neyts [Belgique] ; Marc Van Ranst [Belgique]Source :
- International Journal of Infectious Diseases [ 1201-9712 ] ; 2005.
Descripteurs français
- KwdFr :
- ARN viral (génétique), Animaux, Cellules Vero, Coronavirus (génétique), Coronavirus (isolement et purification), Coronavirus humain 229E (génétique), Coronavirus humain 229E (isolement et purification), Coronavirus humain OC43 (génétique), Coronavirus humain OC43 (isolement et purification), RNA replicase, Reproductibilité des résultats, Réaction de polymérisation en chaîne (), Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : ARN viral, Coronavirus, Coronavirus humain 229E, Coronavirus humain OC43.
- isolement et purification : Coronavirus, Coronavirus humain 229E, Coronavirus humain OC43.
- virologie : Syndrome respiratoire aigu sévère.
- Animaux, Cellules Vero, RNA replicase, Reproductibilité des résultats, Réaction de polymérisation en chaîne, Sensibilité et spécificité.
English descriptors
- KwdEn :
- Animals, Chlorocebus aethiops, Coronavirus (genetics), Coronavirus (isolation & purification), Coronavirus 229E, Human (genetics), Coronavirus 229E, Human (isolation & purification), Coronavirus OC43, Human (genetics), Coronavirus OC43, Human (isolation & purification), Polymerase Chain Reaction (methods), RNA Replicase, RNA, Viral (genetics), Reproducibility of Results, Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Vero Cells.
- MESH :
- chemical , genetics : RNA, Viral.
- chemical : RNA Replicase.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : Coronavirus, Coronavirus 229E, Human, Coronavirus OC43, Human.
- isolation & purification : Coronavirus, Coronavirus 229E, Human, Coronavirus OC43, Human.
- methods : Polymerase Chain Reaction.
- virology : Severe Acute Respiratory Syndrome.
- Animals, Chlorocebus aethiops, Reproducibility of Results, Sensitivity and Specificity, Vero Cells.
Abstract
Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first occurred in humans in the People's Republic of China in November 2002 and has subsequently spread worldwide. A novel virus belonging to the
A real-time quantitative RT-PCR was designed in the nsp11 region of the replicase 1B domain of the SARS-coronavirus (SARS-CoV) genome. To evaluate this quantitative RT-PCR, cRNA standards were constructed by in vitro transcription of SARS-CoV Frankfurt 1 RNA using T7 RNA polymerase, followed by real-time RT-PCR. The assay allowed quantitation over a range of 102 to 108 RNA copies per reaction.
Extrapolated to clinical samples, this novel assay has a detection range of 104 to 1010 copies of viral genome equivalents per millilitre. In comparison to the current de facto cRNA Artus Biotech standard, the in-house cRNA standard gives a 100-fold higher absolute quantity, suggesting a possible underestimation of the viral load when using the Artus Biotech standard.
Url:
DOI: 10.1016/j.ijid.2005.02.003
PubMed: 16023880
PubMed Central: 7110610
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PMC:7110610Le document en format XML
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xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Maes, Piet" sort="Maes, Piet" uniqKey="Maes P" first="Piet" last="Maes">Piet Maes</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Duson, Griet" sort="Duson, Griet" uniqKey="Duson G" first="Griet" last="Duson">Griet Duson</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Neyts, Johan" sort="Neyts, Johan" uniqKey="Neyts J" first="Johan" last="Neyts">Johan Neyts</name><affiliation wicri:level="1"><nlm:aff id="aff2">Laboratory of Virology and Chemotherapy, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Virology and Chemotherapy, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven</wicri:regionArea><wicri:noRegion>University of Leuven</wicri:noRegion></affiliation></author><author><name sortKey="Van Ranst, Marc" sort="Van Ranst, Marc" uniqKey="Van Ranst M" first="Marc" last="Van Ranst">Marc Van Ranst</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation><affiliation wicri:level="1"><nlm:aff id="aff3">Laboratory of Clinical Virology, University Hospital Gasthuisberg, University of Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical Virology, University Hospital Gasthuisberg, University of Leuven</wicri:regionArea><wicri:noRegion>University of Leuven</wicri:noRegion></affiliation></author></analytic><series><title level="j">International Journal of Infectious Diseases</title><idno type="ISSN">1201-9712</idno><idno type="eISSN">1878-3511</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Coronavirus (genetics)</term><term>Coronavirus (isolation & purification)</term><term>Coronavirus 229E, Human (genetics)</term><term>Coronavirus 229E, Human (isolation & purification)</term><term>Coronavirus OC43, Human (genetics)</term><term>Coronavirus OC43, Human (isolation & purification)</term><term>Polymerase Chain Reaction (methods)</term><term>RNA Replicase</term><term>RNA, Viral (genetics)</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Vero Cells</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (génétique)</term><term>Animaux</term><term>Cellules Vero</term><term>Coronavirus (génétique)</term><term>Coronavirus (isolement et purification)</term><term>Coronavirus humain 229E (génétique)</term><term>Coronavirus humain 229E (isolement et purification)</term><term>Coronavirus humain OC43 (génétique)</term><term>Coronavirus humain OC43 (isolement et purification)</term><term>RNA replicase</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaîne ()</term><term>Sensibilité et spécificité</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>RNA Replicase</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>Coronavirus</term><term>Coronavirus 229E, Human</term><term>Coronavirus OC43, Human</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN viral</term><term>Coronavirus</term><term>Coronavirus humain 229E</term><term>Coronavirus humain OC43</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Coronavirus</term><term>Coronavirus 229E, Human</term><term>Coronavirus OC43, Human</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Coronavirus</term><term>Coronavirus humain 229E</term><term>Coronavirus humain OC43</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Vero Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>RNA replicase</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaîne</term><term>Sensibilité et spécificité</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>Summary</title><sec><title>Introduction</title><p>Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first occurred in humans in the People's Republic of China in November 2002 and has subsequently spread worldwide. A novel virus belonging to the <italic>Coronaviridae</italic> family has been identified as the cause of this pulmonary disease. The severity of the disease combined with its rapid spread requires the development of fast and sensitive diagnostic assays.</p></sec><sec><title>Results</title><p>A real-time quantitative RT-PCR was designed in the nsp11 region of the replicase 1B domain of the SARS-coronavirus (SARS-CoV) genome. To evaluate this quantitative RT-PCR, cRNA standards were constructed by in vitro transcription of SARS-CoV Frankfurt 1 RNA using T7 RNA polymerase, followed by real-time RT-PCR. The assay allowed quantitation over a range of 10<sup>2</sup> to 10<sup>8</sup> RNA copies per reaction.</p></sec><sec><title>Conclusions</title><p>Extrapolated to clinical samples, this novel assay has a detection range of 10<sup>4</sup> to 10<sup>10</sup> copies of viral genome equivalents per millilitre. In comparison to the current de facto cRNA Artus Biotech standard, the in-house cRNA standard gives a 100-fold higher absolute quantity, suggesting a possible underestimation of the viral load when using the Artus Biotech standard.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Peiris, J S" uniqKey="Peiris J">J.S. Peiris</name></author><author><name sortKey="Guan, Y" uniqKey="Guan Y">Y. Guan</name></author><author><name sortKey="Yuen, K Y" uniqKey="Yuen K">K.Y. Yuen</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, H R" uniqKey="Brodt H">H.R. 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Cinatl</name></author></analytic></biblStruct></listBibl></div1></back></TEI><double pmid="16023880"><pmc><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR</title><author><name sortKey="Keyaerts, Els" sort="Keyaerts, Els" uniqKey="Keyaerts E" first="Els" last="Keyaerts">Els Keyaerts</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Vijgen, Leen" sort="Vijgen, Leen" uniqKey="Vijgen L" first="Leen" last="Vijgen">Leen Vijgen</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Maes, Piet" sort="Maes, Piet" uniqKey="Maes P" first="Piet" last="Maes">Piet Maes</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Duson, Griet" sort="Duson, Griet" uniqKey="Duson G" first="Griet" last="Duson">Griet Duson</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Neyts, Johan" sort="Neyts, Johan" uniqKey="Neyts J" first="Johan" last="Neyts">Johan Neyts</name><affiliation wicri:level="1"><nlm:aff id="aff2">Laboratory of Virology and Chemotherapy, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Virology and Chemotherapy, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven</wicri:regionArea><wicri:noRegion>University of Leuven</wicri:noRegion></affiliation></author><author><name sortKey="Van Ranst, Marc" sort="Van Ranst, Marc" uniqKey="Van Ranst M" first="Marc" last="Van Ranst">Marc Van Ranst</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation><affiliation wicri:level="1"><nlm:aff id="aff3">Laboratory of Clinical Virology, University Hospital Gasthuisberg, University of Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical Virology, University Hospital Gasthuisberg, University of Leuven</wicri:regionArea><wicri:noRegion>University of Leuven</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">16023880</idno><idno type="pmc">7110610</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7110610</idno><idno type="RBID">PMC:7110610</idno><idno type="doi">10.1016/j.ijid.2005.02.003</idno><date when="2005">2005</date><idno type="wicri:Area/Pmc/Corpus">001086</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001086</idno><idno type="wicri:Area/Pmc/Curation">001086</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">001086</idno><idno type="wicri:Area/Pmc/Checkpoint">001189</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">001189</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR</title><author><name sortKey="Keyaerts, Els" sort="Keyaerts, Els" uniqKey="Keyaerts E" first="Els" last="Keyaerts">Els Keyaerts</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Vijgen, Leen" sort="Vijgen, Leen" uniqKey="Vijgen L" first="Leen" last="Vijgen">Leen Vijgen</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Maes, Piet" sort="Maes, Piet" uniqKey="Maes P" first="Piet" last="Maes">Piet Maes</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Duson, Griet" sort="Duson, Griet" uniqKey="Duson G" first="Griet" last="Duson">Griet Duson</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation></author><author><name sortKey="Neyts, Johan" sort="Neyts, Johan" uniqKey="Neyts J" first="Johan" last="Neyts">Johan Neyts</name><affiliation wicri:level="1"><nlm:aff id="aff2">Laboratory of Virology and Chemotherapy, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Virology and Chemotherapy, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven</wicri:regionArea><wicri:noRegion>University of Leuven</wicri:noRegion></affiliation></author><author><name sortKey="Van Ranst, Marc" sort="Van Ranst, Marc" uniqKey="Van Ranst M" first="Marc" last="Van Ranst">Marc Van Ranst</name><affiliation wicri:level="3"><nlm:aff id="aff1">Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, Minderbroedersstraat 10, 3000 Leuven</wicri:regionArea><placeName><region type="province" nuts="2">Province du Brabant flamand</region><settlement type="city">Louvain</settlement></placeName></affiliation><affiliation wicri:level="1"><nlm:aff id="aff3">Laboratory of Clinical Virology, University Hospital Gasthuisberg, University of Leuven, Belgium</nlm:aff><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical Virology, University Hospital Gasthuisberg, University of Leuven</wicri:regionArea><wicri:noRegion>University of Leuven</wicri:noRegion></affiliation></author></analytic><series><title level="j">International Journal of Infectious Diseases</title><idno type="ISSN">1201-9712</idno><idno type="eISSN">1878-3511</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><title>Summary</title><sec><title>Introduction</title><p>Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first occurred in humans in the People's Republic of China in November 2002 and has subsequently spread worldwide. A novel virus belonging to the <italic>Coronaviridae</italic> family has been identified as the cause of this pulmonary disease. The severity of the disease combined with its rapid spread requires the development of fast and sensitive diagnostic assays.</p></sec><sec><title>Results</title><p>A real-time quantitative RT-PCR was designed in the nsp11 region of the replicase 1B domain of the SARS-coronavirus (SARS-CoV) genome. To evaluate this quantitative RT-PCR, cRNA standards were constructed by in vitro transcription of SARS-CoV Frankfurt 1 RNA using T7 RNA polymerase, followed by real-time RT-PCR. The assay allowed quantitation over a range of 10<sup>2</sup> to 10<sup>8</sup> RNA copies per reaction.</p></sec><sec><title>Conclusions</title><p>Extrapolated to clinical samples, this novel assay has a detection range of 10<sup>4</sup> to 10<sup>10</sup> copies of viral genome equivalents per millilitre. 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Neyts</name></author><author><name sortKey="Van Ranst, M" uniqKey="Van Ranst M">M. Van Ranst</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Yamamoto, N" uniqKey="Yamamoto N">N. Yamamoto</name></author><author><name sortKey="Yang, R" uniqKey="Yang R">R. Yang</name></author><author><name sortKey="Yoshinaka, Y" uniqKey="Yoshinaka Y">Y. Yoshinaka</name></author><author><name sortKey="Amari, S" uniqKey="Amari S">S. Amari</name></author><author><name sortKey="Nakano, T" uniqKey="Nakano T">T. Nakano</name></author><author><name sortKey="Cinatl, J" uniqKey="Cinatl J">J. Cinatl</name></author></analytic></biblStruct></listBibl></div1></back></TEI></pmc><pubmed><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR.</title><author><name sortKey="Keyaerts, Els" sort="Keyaerts, Els" uniqKey="Keyaerts E" first="Els" last="Keyaerts">Els Keyaerts</name><affiliation wicri:level="1"><nlm:affiliation>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, University Hospital Gasthuisberg, Leuven, Belgium.</nlm:affiliation><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, University Hospital Gasthuisberg, Leuven</wicri:regionArea><wicri:noRegion>Leuven</wicri:noRegion></affiliation></author><author><name sortKey="Vijgen, Leen" sort="Vijgen, Leen" uniqKey="Vijgen L" first="Leen" last="Vijgen">Leen Vijgen</name></author><author><name sortKey="Maes, Piet" sort="Maes, Piet" uniqKey="Maes P" first="Piet" last="Maes">Piet Maes</name></author><author><name sortKey="Duson, Griet" sort="Duson, Griet" uniqKey="Duson G" first="Griet" last="Duson">Griet Duson</name></author><author><name sortKey="Neyts, Johan" sort="Neyts, Johan" uniqKey="Neyts J" first="Johan" last="Neyts">Johan Neyts</name></author><author><name sortKey="Van Ranst, Marc" sort="Van Ranst, Marc" uniqKey="Van Ranst M" first="Marc" last="Van Ranst">Marc Van Ranst</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2006">2006</date><idno type="RBID">pubmed:16023880</idno><idno type="pmid">16023880</idno><idno type="doi">10.1016/j.ijid.2005.02.003</idno><idno type="wicri:Area/PubMed/Corpus">002637</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002637</idno><idno type="wicri:Area/PubMed/Curation">002637</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002637</idno><idno type="wicri:Area/PubMed/Checkpoint">001F28</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001F28</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR.</title><author><name sortKey="Keyaerts, Els" sort="Keyaerts, Els" uniqKey="Keyaerts E" first="Els" last="Keyaerts">Els Keyaerts</name><affiliation wicri:level="1"><nlm:affiliation>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, University Hospital Gasthuisberg, Leuven, Belgium.</nlm:affiliation><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Clinical and Epidemiological Virology, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven, University Hospital Gasthuisberg, Leuven</wicri:regionArea><wicri:noRegion>Leuven</wicri:noRegion></affiliation></author><author><name sortKey="Vijgen, Leen" sort="Vijgen, Leen" uniqKey="Vijgen L" first="Leen" last="Vijgen">Leen Vijgen</name></author><author><name sortKey="Maes, Piet" sort="Maes, Piet" uniqKey="Maes P" first="Piet" last="Maes">Piet Maes</name></author><author><name sortKey="Duson, Griet" sort="Duson, Griet" uniqKey="Duson G" first="Griet" last="Duson">Griet Duson</name></author><author><name sortKey="Neyts, Johan" sort="Neyts, Johan" uniqKey="Neyts J" first="Johan" last="Neyts">Johan Neyts</name></author><author><name sortKey="Van Ranst, Marc" sort="Van Ranst, Marc" uniqKey="Van Ranst M" first="Marc" last="Van Ranst">Marc Van Ranst</name></author></analytic><series><title level="j">International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases</title><idno type="ISSN">1201-9712</idno><imprint><date when="2006" type="published">2006</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Coronavirus (genetics)</term><term>Coronavirus (isolation & purification)</term><term>Coronavirus 229E, Human (genetics)</term><term>Coronavirus 229E, Human (isolation & purification)</term><term>Coronavirus OC43, Human (genetics)</term><term>Coronavirus OC43, Human (isolation & purification)</term><term>Polymerase Chain Reaction (methods)</term><term>RNA Replicase</term><term>RNA, Viral (genetics)</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Vero Cells</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (génétique)</term><term>Animaux</term><term>Cellules Vero</term><term>Coronavirus (génétique)</term><term>Coronavirus (isolement et purification)</term><term>Coronavirus humain 229E (génétique)</term><term>Coronavirus humain 229E (isolement et purification)</term><term>Coronavirus humain OC43 (génétique)</term><term>Coronavirus humain OC43 (isolement et purification)</term><term>RNA replicase</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaîne ()</term><term>Sensibilité et spécificité</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>RNA Replicase</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>Coronavirus</term><term>Coronavirus 229E, Human</term><term>Coronavirus OC43, Human</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN viral</term><term>Coronavirus</term><term>Coronavirus humain 229E</term><term>Coronavirus humain OC43</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Coronavirus</term><term>Coronavirus 229E, Human</term><term>Coronavirus OC43, Human</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Coronavirus</term><term>Coronavirus humain 229E</term><term>Coronavirus humain OC43</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Vero Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>RNA replicase</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaîne</term><term>Sensibilité et spécificité</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first occurred in humans in the People's Republic of China in November 2002 and has subsequently spread worldwide. A novel virus belonging to the Coronaviridae family has been identified as the cause of this pulmonary disease. The severity of the disease combined with its rapid spread requires the development of fast and sensitive diagnostic assays.</div></front></TEI></pubmed></double></record>Pour manipuler ce document sous Unix (Dilib)
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