Development of a new detection tool by real time PCR for the detection of Middle East Respiratory Syndrome human Coronavirus (MERS-HCoV) combining specific primers, probe and a RNA internal control ready to use premix
Identifieur interne : 000598 ( Pmc/Corpus ); précédent : 000597; suivant : 000599Development of a new detection tool by real time PCR for the detection of Middle East Respiratory Syndrome human Coronavirus (MERS-HCoV) combining specific primers, probe and a RNA internal control ready to use premix
Auteurs : M. Vignoles ; P. Marechal ; M. Dube ; C. Barranger ; D. Heckel ; M. JoannesSource :
- Journal of Clinical Virology [ 1386-6532 ] ; 2015.
Url:
DOI: 10.1016/j.jcv.2015.07.140
PubMed: NONE
PubMed Central: 7128632
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PMC:7128632Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development of a new detection tool by real time PCR for the detection of Middle East Respiratory Syndrome human Coronavirus (MERS-HCoV) combining specific primers, probe and a RNA internal control ready to use premix</title><author><name sortKey="Vignoles, M" sort="Vignoles, M" uniqKey="Vignoles M" first="M." last="Vignoles">M. Vignoles</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></author><author><name sortKey="Marechal, P" sort="Marechal, P" uniqKey="Marechal P" first="P." last="Marechal">P. Marechal</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></author><author><name sortKey="Dube, M" sort="Dube, M" uniqKey="Dube M" first="M." last="Dube">M. Dube</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></author><author><name sortKey="Barranger, C" sort="Barranger, C" uniqKey="Barranger C" first="C." last="Barranger">C. Barranger</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></author><author><name sortKey="Heckel, D" sort="Heckel, D" uniqKey="Heckel D" first="D." last="Heckel">D. Heckel</name><affiliation><nlm:aff id="aff0010">bioMérieux, Grenoble, France</nlm:aff></affiliation></author><author><name sortKey="Joannes, M" sort="Joannes, M" uniqKey="Joannes M" first="M." last="Joannes">M. Joannes</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmc">7128632</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7128632</idno><idno type="RBID">PMC:7128632</idno><idno type="doi">10.1016/j.jcv.2015.07.140</idno><idno type="pmid">NONE</idno><date when="2015">2015</date><idno type="wicri:Area/Pmc/Corpus">000598</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000598</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Development of a new detection tool by real time PCR for the detection of Middle East Respiratory Syndrome human Coronavirus (MERS-HCoV) combining specific primers, probe and a RNA internal control ready to use premix</title><author><name sortKey="Vignoles, M" sort="Vignoles, M" uniqKey="Vignoles M" first="M." last="Vignoles">M. Vignoles</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></author><author><name sortKey="Marechal, P" sort="Marechal, P" uniqKey="Marechal P" first="P." last="Marechal">P. Marechal</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></author><author><name sortKey="Dube, M" sort="Dube, M" uniqKey="Dube M" first="M." last="Dube">M. Dube</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></author><author><name sortKey="Barranger, C" sort="Barranger, C" uniqKey="Barranger C" first="C." last="Barranger">C. Barranger</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></author><author><name sortKey="Heckel, D" sort="Heckel, D" uniqKey="Heckel D" first="D." last="Heckel">D. Heckel</name><affiliation><nlm:aff id="aff0010">bioMérieux, Grenoble, France</nlm:aff></affiliation></author><author><name sortKey="Joannes, M" sort="Joannes, M" uniqKey="Joannes M" first="M." last="Joannes">M. Joannes</name><affiliation><nlm:aff id="aff0005">bioMérieux, Verniolle, France</nlm:aff></affiliation></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="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader></TEI><pmc article-type="abstract"><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>Published by Elsevier B.V.</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmc">7128632</article-id><article-id pub-id-type="publisher-id">S1386-6532(15)00440-0</article-id><article-id pub-id-type="doi">10.1016/j.jcv.2015.07.140</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Development of a new detection tool by real time PCR for the detection of Middle East Respiratory Syndrome human Coronavirus (MERS-HCoV) combining specific primers, probe and a RNA internal control ready to use premix</article-title></title-group><contrib-group><contrib contrib-type="author" id="aut0005"><name><surname>Vignoles</surname><given-names>M.</given-names></name><xref rid="aff0005" ref-type="aff">1</xref><xref rid="cor0005" ref-type="corresp">⁎</xref></contrib><contrib contrib-type="author" id="aut0010"><name><surname>Marechal</surname><given-names>P.</given-names></name><xref rid="aff0005" ref-type="aff">1</xref></contrib><contrib contrib-type="author" id="aut0015"><name><surname>Dube</surname><given-names>M.</given-names></name><xref rid="aff0005" ref-type="aff">1</xref></contrib><contrib contrib-type="author" id="aut0020"><name><surname>Barranger</surname><given-names>C.</given-names></name><xref rid="aff0005" ref-type="aff">1</xref></contrib><contrib contrib-type="author" id="aut0025"><name><surname>Heckel</surname><given-names>D.</given-names></name><xref rid="aff0010" ref-type="aff">2</xref></contrib><contrib contrib-type="author" id="aut0030"><name><surname>Joannes</surname><given-names>M.</given-names></name><xref rid="aff0005" ref-type="aff">1</xref></contrib></contrib-group><aff id="aff0005"><label>1</label>bioMérieux, Verniolle, France</aff><aff id="aff0010"><label>2</label>bioMérieux, Grenoble, France</aff><author-notes><corresp id="cor0005"><label>⁎</label>Corresponding author.</corresp></author-notes><pub-date pub-type="pmc-release"><day>18</day><month>8</month><year>2015</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>9</month><year>2015</year></pub-date><pub-date pub-type="epub"><day>18</day><month>8</month><year>2015</year></pub-date><volume>70</volume><fpage>S59</fpage><lpage>S59</lpage><permissions><copyright-statement>Copyright © 2015 Published by Elsevier B.V.</copyright-statement><copyright-year>2015</copyright-year><copyright-holder></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></article-meta></front><body><p id="par0005"><bold>Background:</bold> Since the emergence of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) in 2012 in the Arabian Peninsula, many questions remain on modes of transmission and sources of virus. The MERS-HCoV causes severe respiratory illness. The epidemic origins are uncertain but it is probably a zoonosis. The reservoirs could be the bat or the camel. In 2015-04-29, the MERS-HCoV is responsible of 1110 laboratory confirmed cases and 442 deaths (WHO). In outbreak situations, especially with emerging organisms causing severe human diseases, it is important to develop quickly a test to detect the virus involved. bioMérieux developed a real time PCR assay for the rapid detection of MERS-HCoV combining specific primers, probe and a RNA internal control ready-to-use premix. The results of the analytical sensitivity, the precision determination and the exclusivity study are presented.</p><p id="par0010"><bold>Methods:</bold> A set of primers and probe (Mers-hCoV primers r-gene<sup>®</sup> #20-010 and Mers-hCoV probe r-gene<sup>®</sup> #20-011, bioMérieux) were designed on the S gene, coding for the spike structural protein. This set was evaluated in combination with a ready-to-use premix for RNA internal control to constitute a duplex RT PCR assay. The internal control, added before the extraction step, allows to check simultaneously extraction efficiency and presence of inhibitors. Extractions were performed on NucliSENS<sup>®</sup> easyMAG<sup>®</sup> (bioMérieux) followed by amplifications on 7500 Fast Real-Time PCR System (Applied Biosystems<sup>®</sup>). The analytical sensitivity was performed on 20 replicates of a wide dilution range of In Vitro transcripts. The precision determination was determined on 18 replicates of 3 points corresponding at 40, 20 and 8 times LoD 95%. The exclusivity study was carried out on major human respiratory viruses.</p><p id="par0015"><bold>Results:</bold> The analytical sensitivity was determined by Probit analysis (MiniTab). The LoD (95% hit rate detection) was 2.89 log copies/mL of In Vitro transcripts. The coefficients of variation obtained in the precision test were ranged between 1.28% and 2.26% for the MERS-HCoV and between 1.95% and 2.39% for the internal control. The exclusivity study was performed with the major human respiratory viruses including other human coronaviruses, and no cross-reaction was observed.</p><p id="par0020"><bold>Conclusion:</bold> The high quality in analytical sensitivity (2.89) and robustness (1.28–2.39%) were demonstrated. The combination of the MersCoV primers and probe in association with the RNA internal control ready-to-use premix provides a useful tool for the detection of MERS-HCoV virus.</p></body></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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