Clinical evaluation of real-time PCR assays for rapid diagnosis of SARS coronavirus during outbreak and post-epidemic periods
Identifieur interne : 005855 ( Main/Merge ); précédent : 005854; suivant : 005856Clinical evaluation of real-time PCR assays for rapid diagnosis of SARS coronavirus during outbreak and post-epidemic periods
Auteurs : W. C. Yam ; K. H. Chan ; K. H. Chow ; L. L. M. Poon ; H. Y. Lam ; K. Y. Yuen ; W. H. Seto ; J. S. M. PeirisSource :
- Journal of Clinical Virology [ 1386-6532 ] ; 2004.
Descripteurs français
- KwdFr :
- Facteurs temps, Flambées de maladies, Humains, RT-PCR, Réaction de polymérisation en chaîne (), Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Syndrome respiratoire aigu sévère (épidémiologie), Trousses de réactifs pour diagnostic, Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Virus du SRAS.
- isolement et purification : Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- épidémiologie : Syndrome respiratoire aigu sévère.
- Facteurs temps, Flambées de maladies, Humains, RT-PCR, Réaction de polymérisation en chaîne, Sensibilité et spécificité, Trousses de réactifs pour diagnostic.
English descriptors
- KwdEn :
- Disease Outbreaks, Humans, Polymerase Chain Reaction (methods), Reagent Kits, Diagnostic, Reverse Transcriptase Polymerase Chain Reaction, SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (epidemiology), Severe Acute Respiratory Syndrome (virology), Time Factors.
- MESH :
- chemical : Reagent Kits, Diagnostic.
- diagnosis : Severe Acute Respiratory Syndrome.
- epidemiology : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Polymerase Chain Reaction.
- virology : Severe Acute Respiratory Syndrome.
- Disease Outbreaks, Humans, Reverse Transcriptase Polymerase Chain Reaction, Sensitivity and Specificity, Time Factors.
Abstract
The protocols of WHO network laboratories facilitated development of rapid diagnosis for SARS coronavirus (CoV) using reverse transcription (RT)-PCR assays. However, several reports have shown that conventional and real-time PCR assays were very specific for SARS CoV but lack sensitivity depending on the assay, specimen, and time course of disease.
To evaluate an automatic nucleic acid extraction system and two standardized real-time PCR assays for rapid diagnosis of SARS CoV during outbreak and post-epidemic periods in Hong Kong.
Specimens from clinically suspected SARS patients collected during outbreak and post-epidemic periods were tested by an automatic nucleic acid extraction system followed by our first generation conventional RT-PCR and two standardized real-time PCR assays (Artus GmbH, Germany and Roche Diagnostics, Germany). Paired serum samples were assayed for increasing titer against SARS CoV.
In the SARS epidemic, Artus and Roche PCR assays exhibited sensitivities of 87% and 85% for respiratory specimens (
This study highlights the high throughput and performance of automatic RNA extraction in coordination with standardized real-time PCR assays suitable for large-scale routine diagnosis in case of future SARS epidemic. As no SARS CoV was detected among specimens collected during post-epidemic period, the positive predictive value of real-time PCR assays for detection of SARS CoV during low epidemic requires further evaluation.
Url:
DOI: 10.1016/j.jcv.2004.09.029
PubMed: 15797361
PubMed Central: 7108323
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PMC:7108323Le document en format XML
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H." last="Seto">W. H. Seto</name></author><author><name sortKey="Peiris, J S M" sort="Peiris, J S M" uniqKey="Peiris J" first="J. S. M." last="Peiris">J. S. M. Peiris</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">15797361</idno><idno type="pmc">7108323</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7108323</idno><idno type="RBID">PMC:7108323</idno><idno type="doi">10.1016/j.jcv.2004.09.029</idno><date when="2004">2004</date><idno type="wicri:Area/Pmc/Corpus">001117</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001117</idno><idno type="wicri:Area/Pmc/Curation">001117</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">001117</idno><idno type="wicri:Area/Pmc/Checkpoint">001707</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">001707</idno><idno type="wicri:source">PubMed</idno><idno type="RBID">pubmed:15797361</idno><idno type="wicri:Area/PubMed/Corpus">002813</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002813</idno><idno type="wicri:Area/PubMed/Curation">002813</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002813</idno><idno type="wicri:Area/PubMed/Checkpoint">002793</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002793</idno><idno type="wicri:Area/Ncbi/Merge">000E69</idno><idno type="wicri:Area/Ncbi/Curation">000E69</idno><idno type="wicri:Area/Ncbi/Checkpoint">000E69</idno><idno type="wicri:doubleKey">1386-6532:2004:Yam W:clinical:evaluation:of</idno><idno type="wicri:Area/Main/Merge">005855</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Clinical evaluation of real-time PCR assays for rapid diagnosis of SARS coronavirus during outbreak and post-epidemic periods</title><author><name sortKey="Yam, W C" sort="Yam, W C" uniqKey="Yam W" first="W. C." last="Yam">W. C. Yam</name></author><author><name sortKey="Chan, K H" sort="Chan, K H" uniqKey="Chan K" first="K. H." last="Chan">K. H. Chan</name></author><author><name sortKey="Chow, K H" sort="Chow, K H" uniqKey="Chow K" first="K. H." last="Chow">K. H. Chow</name></author><author><name sortKey="Poon, L L M" sort="Poon, L L M" uniqKey="Poon L" first="L. L. M." last="Poon">L. L. M. Poon</name></author><author><name sortKey="Lam, H Y" sort="Lam, H Y" uniqKey="Lam H" first="H. Y." last="Lam">H. Y. Lam</name></author><author><name sortKey="Yuen, K Y" sort="Yuen, K Y" uniqKey="Yuen K" first="K. Y." last="Yuen">K. Y. Yuen</name></author><author><name sortKey="Seto, W H" sort="Seto, W H" uniqKey="Seto W" first="W. H." last="Seto">W. H. Seto</name></author><author><name sortKey="Peiris, J S M" sort="Peiris, J S M" uniqKey="Peiris J" first="J. S. M." last="Peiris">J. S. M. Peiris</name></author></analytic><series><title level="j">Journal of Clinical Virology</title><idno type="ISSN">1386-6532</idno><idno type="eISSN">1873-5967</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Disease Outbreaks</term><term>Humans</term><term>Polymerase Chain Reaction (methods)</term><term>Reagent Kits, Diagnostic</term><term>Reverse Transcriptase 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 (epidemiology)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Time Factors</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Facteurs temps</term><term>Flambées de maladies</term><term>Humains</term><term>RT-PCR</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><term>Syndrome respiratoire aigu sévère (épidémiologie)</term><term>Trousses de réactifs pour diagnostic</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Reagent Kits, Diagnostic</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="epidemiology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><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>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" qualifier="épidémiologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Disease Outbreaks</term><term>Humans</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Sensitivity and Specificity</term><term>Time Factors</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Facteurs temps</term><term>Flambées de maladies</term><term>Humains</term><term>RT-PCR</term><term>Réaction de polymérisation en chaîne</term><term>Sensibilité et spécificité</term><term>Trousses de réactifs pour diagnostic</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background:</title><p>The protocols of WHO network laboratories facilitated development of rapid diagnosis for SARS coronavirus (CoV) using reverse transcription (RT)-PCR assays. However, several reports have shown that conventional and real-time PCR assays were very specific for SARS CoV but lack sensitivity depending on the assay, specimen, and time course of disease.</p></sec><sec><title>Objective:</title><p>To evaluate an automatic nucleic acid extraction system and two standardized real-time PCR assays for rapid diagnosis of SARS CoV during outbreak and post-epidemic periods in Hong Kong.</p></sec><sec><title>Study design:</title><p>Specimens from clinically suspected SARS patients collected during outbreak and post-epidemic periods were tested by an automatic nucleic acid extraction system followed by our first generation conventional RT-PCR and two standardized real-time PCR assays (Artus GmbH, Germany and Roche Diagnostics, Germany). Paired serum samples were assayed for increasing titer against SARS CoV.</p></sec><sec><title>Results:</title><p>In the SARS epidemic, Artus and Roche PCR assays exhibited sensitivities of 87% and 85% for respiratory specimens (<italic>n</italic> = 64), 91% and 88% for stool (<italic>n</italic> = 44), and 82% for urine (<italic>n</italic> = 29). A specificity of 100% was exhibited by both PCR assays except Artus attained only a 92% specificity for stool. For post-epidemic period, no SARS CoV was identified among 56 respiratory specimens by all PCR assays. Inhibitors to PCR assays were detected at an average rate of 7–8% among 202 clinical specimens.</p></sec><sec><title>Conclusion:</title><p>This study highlights the high throughput and performance of automatic RNA extraction in coordination with standardized real-time PCR assays suitable for large-scale routine diagnosis in case of future SARS epidemic. As no SARS CoV was detected among specimens collected during post-epidemic period, the positive predictive value of real-time PCR assays for detection of SARS CoV during low epidemic requires further evaluation.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Chan, K H" uniqKey="Chan K">K.H. Chan</name></author><author><name sortKey="Maldeis, N" uniqKey="Maldeis N">N. Maldeis</name></author><author><name sortKey="Pope, W" uniqKey="Pope W">W. Pope</name></author><author><name sortKey="Yup, A" uniqKey="Yup A">A. Yup</name></author><author><name sortKey="Ozinska, A" uniqKey="Ozinska A">A. Ozinska</name></author><author><name sortKey="Gill, J" uniqKey="Gill J">J. Gill</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Chan, K H" uniqKey="Chan K">K.H. Chan</name></author><author><name sortKey="Poon, L L L M" uniqKey="Poon L">L.L.L.M. Poon</name></author><author><name sortKey="Cheng, V C C" uniqKey="Cheng V">V.C.C. Cheng</name></author><author><name sortKey="Guan, Y" uniqKey="Guan Y">Y. Guan</name></author><author><name sortKey="Hung, I F N" uniqKey="Hung I">I.F.N. Hung</name></author><author><name sortKey="Kong, J" uniqKey="Kong J">J. Kong</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. Brodt</name></author><author><name sortKey="Becker, S" uniqKey="Becker S">S. Becker</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Drosten, C" uniqKey="Drosten C">C. Drosten</name></author><author><name sortKey="Chiu, L" uniqKey="Chiu L">L. Chiu</name></author><author><name sortKey="Panning, M" uniqKey="Panning M">M. Panning</name></author><author><name sortKey="Leong, H N" uniqKey="Leong H">H.N. Leong</name></author><author><name sortKey="Preiser, W" uniqKey="Preiser W">W. Preiser</name></author><author><name sortKey="Tam, J S" uniqKey="Tam J">J.S. Tam</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hourfar, M" uniqKey="Hourfar M">M. Hourfar</name></author><author><name sortKey="Roth, W K" uniqKey="Roth W">W.K. Roth</name></author><author><name sortKey="Seifried, E" uniqKey="Seifried E">E. Seifried</name></author><author><name sortKey="Schmidt, M" uniqKey="Schmidt M">M. Schmidt</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Lee, N" uniqKey="Lee N">N. Lee</name></author><author><name sortKey="Hui, D" uniqKey="Hui D">D. Hui</name></author><author><name sortKey="Wu, A" uniqKey="Wu A">A. Wu</name></author><author><name sortKey="Chan, P" uniqKey="Chan P">P. Chan</name></author><author><name sortKey="Cameron, P" uniqKey="Cameron P">P. Cameron</name></author><author><name sortKey="Joynt, G M" uniqKey="Joynt G">G.M. Joynt</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mazzulli, T" uniqKey="Mazzulli T">T. Mazzulli</name></author><author><name sortKey="Farcas, G A" uniqKey="Farcas G">G.A. Farcas</name></author><author><name sortKey="Poutanen, S M" uniqKey="Poutanen S">S.M. Poutanen</name></author><author><name sortKey="Willey, B M" uniqKey="Willey B">B.M. Willey</name></author><author><name sortKey="Low, D E" uniqKey="Low D">D.E. Low</name></author><author><name sortKey="Butany, J" uniqKey="Butany J">J. Butany</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Peiris, J S M" uniqKey="Peiris J">J.S.M. Peiris</name></author><author><name sortKey="Lai, S T" uniqKey="Lai S">S.T. Lai</name></author><author><name sortKey="Poon, L L M" uniqKey="Poon L">L.L.M. Poon</name></author><author><name sortKey="Guan, Y" uniqKey="Guan Y">Y. Guan</name></author><author><name sortKey="Yam, L Y C" uniqKey="Yam L">L.Y.C. Yam</name></author><author><name sortKey="Lim, W" uniqKey="Lim W">W. Lim</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Peiris, J S" uniqKey="Peiris J">J.S. Peiris</name></author><author><name sortKey="Chu, C M" uniqKey="Chu C">C.M. Chu</name></author><author><name sortKey="Cheng, V C C" uniqKey="Cheng V">V.C.C. Cheng</name></author><author><name sortKey="Chan, K S" uniqKey="Chan K">K.S. Chan</name></author><author><name sortKey="Hung, I F" uniqKey="Hung I">I.F. Hung</name></author><author><name sortKey="Poon, L L" uniqKey="Poon L">L.L. Poon</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Poon, L L M" uniqKey="Poon L">L.L.M. Poon</name></author><author><name sortKey="Wong, O K" uniqKey="Wong O">O.K. Wong</name></author><author><name sortKey="Chan, K H" uniqKey="Chan K">K.H. Chan</name></author><author><name sortKey="Luk, W" uniqKey="Luk W">W. Luk</name></author><author><name sortKey="Yuen, K Y" uniqKey="Yuen K">K.Y. Yuen</name></author><author><name sortKey="Peiris, J S M" uniqKey="Peiris J">J.S.M. Peiris</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Poon, L L M" uniqKey="Poon L">L.L.M. Poon</name></author><author><name sortKey="Chan, K H" uniqKey="Chan K">K.H. Chan</name></author><author><name sortKey="Wong, O K" uniqKey="Wong O">O.K. Wong</name></author><author><name sortKey="Yam, W C" uniqKey="Yam W">W.C. Yam</name></author><author><name sortKey="Yuen, K Y" uniqKey="Yuen K">K.Y. Yuen</name></author><author><name sortKey="Guan, Y" uniqKey="Guan Y">Y. Guan</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Riley, S" uniqKey="Riley S">S. Riley</name></author><author><name sortKey="Fraser, C" uniqKey="Fraser C">C. Fraser</name></author><author><name sortKey="Donnelly, C A" uniqKey="Donnelly C">C.A. Donnelly</name></author><author><name sortKey="Ghani, A C" uniqKey="Ghani A">A.C. Ghani</name></author><author><name sortKey="Abu Raddad, L J" uniqKey="Abu Raddad L">L.J. Abu-Raddad</name></author><author><name sortKey="Hedley, A J" uniqKey="Hedley A">A.J. Hedley</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tsang, K W" uniqKey="Tsang K">K.W. Tsang</name></author><author><name sortKey="Ho, P L" uniqKey="Ho P">P.L. Ho</name></author><author><name sortKey="Ooi, G C" uniqKey="Ooi G">G.C. Ooi</name></author><author><name sortKey="Yee, W K" uniqKey="Yee W">W.K. Yee</name></author><author><name sortKey="Wang, T" uniqKey="Wang T">T. Wang</name></author><author><name sortKey="Chan Yeung, M" uniqKey="Chan Yeung M">M. Chan-Yeung</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Yam, W C" uniqKey="Yam W">W.C. Yam</name></author><author><name sortKey="Chan, K H" uniqKey="Chan K">K.H. Chan</name></author><author><name sortKey="Poon, L L M" uniqKey="Poon L">L.L.M. Poon</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><author><name sortKey="Seto, W H" uniqKey="Seto W">W.H. Seto</name></author></analytic></biblStruct></listBibl></div1></back></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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