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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 : 001118

Clinical 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. Peiris

Source :

RBID : PMC:7108323

Abstract

Background:

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.

Objective:

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.

Study design:

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.

Results:

In the SARS epidemic, Artus and Roche PCR assays exhibited sensitivities of 87% and 85% for respiratory specimens (n = 64), 91% and 88% for stool (n = 44), and 82% for urine (n = 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.

Conclusion:

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:7108323

Le document en format XML

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<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>
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<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>
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<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>
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<title>Results:</title>
<p>In the SARS epidemic, Artus and Roche PCR assays exhibited sensitivities of 87% and 85% for respiratory specimens (
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<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>
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<journal-id journal-id-type="nlm-ta">J Clin Virol</journal-id>
<journal-id journal-id-type="iso-abbrev">J. Clin. Virol</journal-id>
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<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>
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<publisher-name>Elsevier B.V.</publisher-name>
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<article-id pub-id-type="doi">10.1016/j.jcv.2004.09.029</article-id>
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<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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