Sensitive Detection of SARS Coronavirus RNA by a Novel Asymmetric Multiplex Nested RT-PCR Amplification Coupled With Oligonucleotide Microarray Hybridization
Identifieur interne : 001237 ( Pmc/Checkpoint ); précédent : 001236; suivant : 001238Sensitive Detection of SARS Coronavirus RNA by a Novel Asymmetric Multiplex Nested RT-PCR Amplification Coupled With Oligonucleotide Microarray Hybridization
Auteurs :Source :
- Microarrays in Clinical Diagnostics ; 2005.
Abstract
We have developed a sensitive method for the detection of specific genes simultaneously. First, DNA was amplified by a novel asymmetric multiplex PCR with universal primer(s). Second, the 6-carboxytetramethylrhodamine (TAMRA)-labeled PCR products were hybridized specifically with oligonucleotide microarrays. Finally, matched duplexes were detected by using a laser-induced fluorescence scanner. The usefulness of this method was illustrated by analyzing severe acute respiratory syndrome (SARS) coronavirus RNA. The detection limit was 100 copies/µL. The results of the asymmetric multiplex nested reverse transcription-PCR were in agreement with the results of the microarray hybridization; no hybridization signal was lost as happened with applicons from symmetric amplifications. This reliable method can be used to the identification of other microorganisms, screening of genetic diseases, and other applications.
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DOI: 10.1385/1-59259-923-0:59
PubMed: 16156097
PubMed Central: 7122606
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Sensitive Detection of SARS Coronavirus RNA by a Novel Asymmetric Multiplex Nested RT-PCR Amplification Coupled With Oligonucleotide Microarray Hybridization</title></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">16156097</idno><idno type="pmc">7122606</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7122606</idno><idno type="RBID">PMC:7122606</idno><idno type="doi">10.1385/1-59259-923-0:59</idno><date when="2005">2005</date><idno type="wicri:Area/Pmc/Corpus">000097</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000097</idno><idno type="wicri:Area/Pmc/Curation">000097</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000097</idno><idno type="wicri:Area/Pmc/Checkpoint">001237</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">001237</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Sensitive Detection of SARS Coronavirus RNA by a Novel Asymmetric Multiplex Nested RT-PCR Amplification Coupled With Oligonucleotide Microarray Hybridization</title></analytic><series><title level="j">Microarrays in Clinical Diagnostics</title><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>We have developed a sensitive method for the detection of specific genes simultaneously. First, DNA was amplified by a novel asymmetric multiplex PCR with universal primer(s). Second, the 6-carboxytetramethylrhodamine (TAMRA)-labeled PCR products were hybridized specifically with oligonucleotide microarrays. Finally, matched duplexes were detected by using a laser-induced fluorescence scanner. The usefulness of this method was illustrated by analyzing severe acute respiratory syndrome (SARS) coronavirus RNA. The detection limit was 10<sup>0</sup> copies/µL. The results of the asymmetric multiplex nested reverse transcription-PCR were in agreement with the results of the microarray hybridization; no hybridization signal was lost as happened with applicons from symmetric amplifications. This reliable method can be used to the identification of other microorganisms, screening of genetic diseases, and other applications.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Chamberlain, J S" uniqKey="Chamberlain J">J. S. 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<front><journal-meta><journal-id journal-id-type="publisher-id">978-1-59259-923-3</journal-id><journal-id journal-id-type="doi">10.1385/1592599230</journal-id><journal-id journal-id-type="nlm-ta">Microarrays in Clinical Diagnostics</journal-id><journal-title-group><journal-title>Microarrays in Clinical Diagnostics</journal-title></journal-title-group><isbn publication-format="print">978-1-58829-394-7</isbn><isbn publication-format="electronic">978-1-59259-923-3</isbn></journal-meta><article-meta><article-id pub-id-type="pmid">16156097</article-id><article-id pub-id-type="pmc">7122606</article-id><article-id pub-id-type="publisher-id">3</article-id><article-id pub-id-type="doi">10.1385/1-59259-923-0:59</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Sensitive Detection of SARS Coronavirus RNA by a Novel Asymmetric Multiplex Nested RT-PCR Amplification Coupled With Oligonucleotide Microarray Hybridization</article-title></title-group><contrib-group content-type="book editors"><contrib contrib-type="editor"><name><surname>Joos</surname><given-names>Thomas O.</given-names></name><degrees>PhD</degrees><xref ref-type="aff" rid="Aff1">1</xref></contrib><contrib contrib-type="editor"><name><surname>Fortina</surname><given-names>Paolo</given-names></name><degrees>MD, PhD</degrees><xref ref-type="aff" rid="Aff2">2</xref></contrib><aff id="Aff1"><label>1</label><institution-wrap><institution-id institution-id-type="GRID">grid.461765.7</institution-id><institution-id institution-id-type="ISNI">0000000094571306</institution-id><institution>Biochemistry Department,</institution><institution>NMI Natural and Medical Sciences Institute at the University of Tuebingen,</institution></institution-wrap>Reutlingen, Germany</aff><aff id="Aff2"><label>2</label><institution-wrap><institution-id institution-id-type="GRID">grid.265008.9</institution-id><institution-id institution-id-type="ISNI">0000000121665843</institution-id><institution>Center for Translational Medicine, Department of Medicine,</institution><institution>Jefferson Medical College,</institution></institution-wrap>Philadelphia, PA</aff></contrib-group><contrib-group><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Zhi-wei</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Zhou</surname><given-names>Yi-ming</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Yan</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Guo</surname><given-names>Yong</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Tao</surname><given-names>Sheng-ce</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Ze</given-names></name><xref ref-type="aff" rid="Aff4">4</xref></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Qiong</given-names></name><xref ref-type="aff" rid="Aff4">4</xref></contrib><contrib contrib-type="author"><name><surname>Cheng</surname><given-names>Jing</given-names></name><xref ref-type="aff" rid="Aff3">3</xref><xref ref-type="aff" rid="Aff4">4</xref></contrib><aff id="Aff3"><label>3</label><institution-wrap><institution-id institution-id-type="GRID">grid.12527.33</institution-id><institution-id institution-id-type="ISNI">0000000106623178</institution-id><institution>State Key Laboratory of Biomembrane and Membrane Biotechnology, Department of Biological Sciences and Biotechnology,</institution><institution>Tsinghua University,</institution></institution-wrap>Beijing, China</aff><aff id="Aff4"><label>4</label>National Engineering Research Center for Beijing Biochip Technology, CapitalBio Corporation, Beijing, China</aff></contrib-group><pub-date pub-type="ppub"><year>2005</year></pub-date><volume>114</volume><fpage>59</fpage><lpage>78</lpage><permissions><copyright-statement>© Humana Press Inc. 2005</copyright-statement><license><license-p>This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.</license-p></license></permissions><abstract id="Abs1"><p>We have developed a sensitive method for the detection of specific genes simultaneously. First, DNA was amplified by a novel asymmetric multiplex PCR with universal primer(s). Second, the 6-carboxytetramethylrhodamine (TAMRA)-labeled PCR products were hybridized specifically with oligonucleotide microarrays. Finally, matched duplexes were detected by using a laser-induced fluorescence scanner. The usefulness of this method was illustrated by analyzing severe acute respiratory syndrome (SARS) coronavirus RNA. The detection limit was 10<sup>0</sup> copies/µL. The results of the asymmetric multiplex nested reverse transcription-PCR were in agreement with the results of the microarray hybridization; no hybridization signal was lost as happened with applicons from symmetric amplifications. This reliable method can be used to the identification of other microorganisms, screening of genetic diseases, and other applications.</p></abstract><kwd-group xml:lang="en"><title>Key Words</title><kwd>Polymerase chain reaction (PCR)</kwd><kwd>multiplex PCR</kwd><kwd>asymmetric PCR</kwd><kwd>universal primer</kwd><kwd>severe acute respiratory syndrome (SARS) coronavirus</kwd><kwd>microarray</kwd></kwd-group><custom-meta-group><custom-meta><meta-name>issue-copyright-statement</meta-name><meta-value>© Humana Press 2005</meta-value></custom-meta></custom-meta-group></article-meta></front></pmc><affiliations><list></list><tree></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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