Clinical evaluation of real-time PCR assays for rapid diagnosis of SARS coronavirus during outbreak and post-epidemic periods
Identifieur interne : 001117 ( Pmc/Curation ); précédent : 001116; suivant : 001118Clinical 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.
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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<author><name sortKey="Yam, W C" sort="Yam, W C" uniqKey="Yam W" first="W. C." last="Yam">W. C. Yam</name>
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<author><name sortKey="Chan, K H" sort="Chan, K H" uniqKey="Chan K" first="K. H." last="Chan">K. H. Chan</name>
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<author><name sortKey="Chow, K H" sort="Chow, K H" uniqKey="Chow K" first="K. H." last="Chow">K. H. Chow</name>
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<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>
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<author><name sortKey="Lam, H Y" sort="Lam, H Y" uniqKey="Lam H" first="H. Y." last="Lam">H. Y. Lam</name>
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<author><name sortKey="Yuen, K Y" sort="Yuen, K Y" uniqKey="Yuen K" first="K. Y." last="Yuen">K. Y. Yuen</name>
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<author><name sortKey="Seto, W H" sort="Seto, W H" uniqKey="Seto W" first="W. H." last="Seto">W. H. Seto</name>
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<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>
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<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>
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<author><name sortKey="Chan, K H" sort="Chan, K H" uniqKey="Chan K" first="K. H." last="Chan">K. H. Chan</name>
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<author><name sortKey="Chow, K H" sort="Chow, K H" uniqKey="Chow K" first="K. H." last="Chow">K. H. Chow</name>
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<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>
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<author><name sortKey="Lam, H Y" sort="Lam, H Y" uniqKey="Lam H" first="H. Y." last="Lam">H. Y. Lam</name>
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<author><name sortKey="Yuen, K Y" sort="Yuen, K Y" uniqKey="Yuen K" first="K. Y." last="Yuen">K. Y. Yuen</name>
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<author><name sortKey="Seto, W H" sort="Seto, W H" uniqKey="Seto W" first="W. H." last="Seto">W. H. Seto</name>
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<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>
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<series><title level="j">Journal of Clinical Virology</title>
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<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>
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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Clin Virol</journal-id>
<journal-id journal-id-type="iso-abbrev">J. Clin. Virol</journal-id>
<journal-title-group><journal-title>Journal of Clinical Virology</journal-title>
</journal-title-group>
<issn pub-type="ppub">1386-6532</issn>
<issn pub-type="epub">1873-5967</issn>
<publisher><publisher-name>Elsevier B.V.</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">15797361</article-id>
<article-id pub-id-type="pmc">7108323</article-id>
<article-id pub-id-type="publisher-id">S1386-6532(04)00286-0</article-id>
<article-id pub-id-type="doi">10.1016/j.jcv.2004.09.029</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Clinical evaluation of real-time PCR assays for rapid diagnosis of SARS coronavirus during outbreak and post-epidemic periods</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Yam</surname>
<given-names>W.C.</given-names>
</name>
<email>wcyam@hku.hk</email>
<xref rid="cor1" ref-type="corresp">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chan</surname>
<given-names>K.H.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Chow</surname>
<given-names>K.H.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Poon</surname>
<given-names>L.L.M.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Lam</surname>
<given-names>H.Y.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Yuen</surname>
<given-names>K.Y.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Seto</surname>
<given-names>W.H.</given-names>
</name>
</contrib>
<contrib contrib-type="author"><name><surname>Peiris</surname>
<given-names>J.S.M.</given-names>
</name>
</contrib>
</contrib-group>
<aff>Department of Microbiology, The University of Hong Kong, University Pathology Building, Queen Mary Hospital Compound, Pokfulam, Hong Kong SAR, PR China</aff>
<author-notes><corresp id="cor1"><label>*</label>
Corresponding author. Tel.: +86 852 28554821; fax: +86 852 28551241. <email>wcyam@hku.hk</email>
</corresp>
</author-notes>
<pub-date pub-type="pmc-release"><day>1</day>
<month>12</month>
<year>2004</year>
</pub-date>
<pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><month>5</month>
<year>2005</year>
</pub-date>
<pub-date pub-type="epub"><day>1</day>
<month>12</month>
<year>2004</year>
</pub-date>
<volume>33</volume>
<issue>1</issue>
<fpage>19</fpage>
<lpage>24</lpage>
<history><date date-type="received"><day>25</day>
<month>3</month>
<year>2004</year>
</date>
<date date-type="rev-recd"><day>14</day>
<month>9</month>
<year>2004</year>
</date>
<date date-type="accepted"><day>24</day>
<month>9</month>
<year>2004</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2004 Elsevier B.V. All rights reserved.</copyright-statement>
<copyright-year>2004</copyright-year>
<copyright-holder>Elsevier B.V.</copyright-holder>
<license><license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p>
</license>
</permissions>
<abstract><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>
</abstract>
<kwd-group><title>Keywords</title>
<kwd>Rapid diagnosis</kwd>
<kwd>SARS coronavirus</kwd>
<kwd>Real-time PCR</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
</record>
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