A one step quantitative RT-PCR for detection of SARS coronavirus with an internal control for PCR inhibitors
Identifieur interne : 005B81 ( Main/Exploration ); précédent : 005B80; suivant : 005B82A one step quantitative RT-PCR for detection of SARS coronavirus with an internal control for PCR inhibitors
Auteurs : Leo L. M. Poon [Hong Kong] ; Bonnie W. Y. Wong [Hong Kong] ; KWOK HUNG CHAN [Hong Kong] ; Cynthia S. W. Leung [Hong Kong] ; KWOK YUNG YUEN [Hong Kong] ; YI GUAN [Hong Kong] ; Joseph S. M. Peiris [Hong Kong]Source :
- Journal of clinical virology [ 1386-6532 ] ; 2004.
Descripteurs français
- KwdFr :
- ADN ribosomique (analyse), ARN ribosomique 18S (génétique), ARN viral (analyse), ARN viral (isolement et purification), Humains, Normes de référence, Partie nasale du pharynx (virologie), RT-PCR (), Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- analyse : ADN ribosomique, ARN viral.
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : ARN ribosomique 18S, Virus du SRAS.
- isolement et purification : ARN viral, Virus du SRAS.
- virologie : Partie nasale du pharynx, Syndrome respiratoire aigu sévère.
- Pascal (Inist)
- Wicri :
- topic : Analyse quantitative.
English descriptors
- KwdEn :
- Check, Coronavirus, DNA, Ribosomal (analysis), Detection, Diagnosis, Early, Humans, Inhibitor, Microbiology, Nasopharynx (virology), Polymerase chain reaction, Quantitative analysis, RNA, Ribosomal, 18S (genetics), RNA, Viral (analysis), RNA, Viral (isolation & purification), Reference Standards, 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 (virology), Severe acute respiratory syndrome, Virology.
- MESH :
- chemical , analysis : DNA, Ribosomal, RNA, Viral.
- chemical , genetics : RNA, Ribosomal, 18S.
- chemical , isolation & purification : RNA, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction.
- virology : Nasopharynx, Severe Acute Respiratory Syndrome.
- Humans, Reference Standards, Sensitivity and Specificity.
Abstract
In this study, we report a one step quantitative RT-PCR assay for severe acute respiratory syndrome (SARS) diagnosis. The overall detection rate for clinical samples collected from Days 1 to 9 of disease onset is 86.2% and the specificity of the assay is 100%. To detect false negative results due to PCR inhibitors or faulty RNA extraction, we have further modified this one step RT-PCR assay for simultaneous detection of severe acute respiratory syndrome (SARS) coronavirus (CoV) and 18S ribosomal RNA (rRNA) as an internal positive control.
Affiliations:
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M." last="Poon">Leo L. M. Poon</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Microbiology, Queen Mary Hospital, University of Hong Kong</s1><s2>Pokfulam SAR</s2><s3>HKG</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Hong Kong</country><wicri:noRegion>Pokfulam SAR</wicri:noRegion></affiliation></author><author><name sortKey="Wong, Bonnie W Y" sort="Wong, Bonnie W Y" uniqKey="Wong B" first="Bonnie W. Y." last="Wong">Bonnie W. Y. Wong</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Microbiology, Queen Mary Hospital, University of Hong Kong</s1><s2>Pokfulam SAR</s2><s3>HKG</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Hong Kong</country><wicri:noRegion>Pokfulam SAR</wicri:noRegion></affiliation></author><author><name sortKey="Kwok Hung Chan" sort="Kwok Hung Chan" uniqKey="Kwok Hung Chan" last="Kwok Hung Chan">KWOK HUNG CHAN</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Department of Microbiology, Queen Mary Hospital</s1><s2>SAR</s2><s3>HKG</s3><sZ>3 aut.</sZ></inist:fA14><country>Hong Kong</country><wicri:noRegion>SAR</wicri:noRegion></affiliation></author><author><name sortKey="Leung, Cynthia S W" sort="Leung, Cynthia S W" uniqKey="Leung C" first="Cynthia S. W." last="Leung">Cynthia S. W. Leung</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Microbiology, Queen Mary Hospital, University of Hong Kong</s1><s2>Pokfulam SAR</s2><s3>HKG</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Hong Kong</country><wicri:noRegion>Pokfulam SAR</wicri:noRegion></affiliation></author><author><name sortKey="Kwok Yung Yuen" sort="Kwok Yung Yuen" uniqKey="Kwok Yung Yuen" last="Kwok Yung Yuen">KWOK YUNG YUEN</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Microbiology, Queen Mary Hospital, University of Hong Kong</s1><s2>Pokfulam SAR</s2><s3>HKG</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Hong Kong</country><wicri:noRegion>Pokfulam SAR</wicri:noRegion></affiliation></author><author><name sortKey="Yi Guan" sort="Yi Guan" uniqKey="Yi Guan" last="Yi Guan">YI GUAN</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Microbiology, Queen Mary Hospital, University of Hong Kong</s1><s2>Pokfulam SAR</s2><s3>HKG</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Hong Kong</country><wicri:noRegion>Pokfulam SAR</wicri:noRegion></affiliation></author><author><name sortKey="Peiris, Joseph S M" sort="Peiris, Joseph S M" uniqKey="Peiris J" first="Joseph S. M." last="Peiris">Joseph S. M. Peiris</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Microbiology, Queen Mary Hospital, University of Hong Kong</s1><s2>Pokfulam SAR</s2><s3>HKG</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Hong Kong</country><wicri:noRegion>Pokfulam SAR</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Journal of clinical virology</title><title level="j" type="abbreviated">J. clin. virol.</title><idno type="ISSN">1386-6532</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of clinical virology</title><title level="j" type="abbreviated">J. clin. virol.</title><idno type="ISSN">1386-6532</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Check</term><term>Coronavirus</term><term>DNA, Ribosomal (analysis)</term><term>Detection</term><term>Diagnosis</term><term>Early</term><term>Humans</term><term>Inhibitor</term><term>Microbiology</term><term>Nasopharynx (virology)</term><term>Polymerase chain reaction</term><term>Quantitative analysis</term><term>RNA, Ribosomal, 18S (genetics)</term><term>RNA, Viral (analysis)</term><term>RNA, Viral (isolation & purification)</term><term>Reference Standards</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 (virology)</term><term>Severe acute respiratory syndrome</term><term>Virology</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN ribosomique (analyse)</term><term>ARN ribosomique 18S (génétique)</term><term>ARN viral (analyse)</term><term>ARN viral (isolement et purification)</term><term>Humains</term><term>Normes de référence</term><term>Partie nasale du pharynx (virologie)</term><term>RT-PCR ()</term><term>Sensibilité et spécificité</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>DNA, Ribosomal</term><term>RNA, Viral</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Ribosomal, 18S</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ADN ribosomique</term><term>ARN viral</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>ARN ribosomique 18S</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>ARN viral</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Reverse Transcriptase Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Partie nasale du pharynx</term><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Nasopharynx</term><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term><term>Reference Standards</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term><term>Analyse quantitative</term><term>Humains</term><term>Normes de référence</term><term>RT-PCR</term><term>Réaction chaîne polymérase RT</term><term>Détection</term><term>Contrôle</term><term>Réaction chaîne polymérase</term><term>Inhibiteur</term><term>Précoce</term><term>Diagnostic</term><term>Sensibilité et spécificité</term><term>Syndrome respiratoire aigu sévère</term><term>Microbiologie</term><term>Virologie</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Analyse quantitative</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">In this study, we report a one step quantitative RT-PCR assay for severe acute respiratory syndrome (SARS) diagnosis. The overall detection rate for clinical samples collected from Days 1 to 9 of disease onset is 86.2% and the specificity of the assay is 100%. To detect false negative results due to PCR inhibitors or faulty RNA extraction, we have further modified this one step RT-PCR assay for simultaneous detection of severe acute respiratory syndrome (SARS) coronavirus (CoV) and 18S ribosomal RNA (rRNA) as an internal positive control.</div></front></TEI><affiliations><list><country><li>Hong Kong</li></country></list><tree><country name="Hong Kong"><noRegion><name sortKey="Poon, Leo L M" sort="Poon, Leo L M" uniqKey="Poon L" first="Leo L. M." last="Poon">Leo L. M. Poon</name></noRegion><name sortKey="Kwok Hung Chan" sort="Kwok Hung Chan" uniqKey="Kwok Hung Chan" last="Kwok Hung Chan">KWOK HUNG CHAN</name><name sortKey="Kwok Yung Yuen" sort="Kwok Yung Yuen" uniqKey="Kwok Yung Yuen" last="Kwok Yung Yuen">KWOK YUNG YUEN</name><name sortKey="Leung, Cynthia S W" sort="Leung, Cynthia S W" uniqKey="Leung C" first="Cynthia S. W." last="Leung">Cynthia S. W. Leung</name><name sortKey="Peiris, Joseph S M" sort="Peiris, Joseph S M" uniqKey="Peiris J" first="Joseph S. M." last="Peiris">Joseph S. M. Peiris</name><name sortKey="Wong, Bonnie W Y" sort="Wong, Bonnie W Y" uniqKey="Wong B" first="Bonnie W. Y." last="Wong">Bonnie W. Y. Wong</name><name sortKey="Yi Guan" sort="Yi Guan" uniqKey="Yi Guan" last="Yi Guan">YI GUAN</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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