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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome Coronavirus</title><author><name sortKey="Thai, Hong Thi Cam" sort="Thai, Hong Thi Cam" uniqKey="Thai H" first="Hong Thi Cam" last="Thai">Hong Thi Cam Thai</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Le, Mai Quynh" sort="Le, Mai Quynh" uniqKey="Le M" first="Mai Quynh" last="Le">Mai Quynh Le</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Vuong, Cuong Duc" sort="Vuong, Cuong Duc" uniqKey="Vuong C" first="Cuong Duc" last="Vuong">Cuong Duc Vuong</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Parida, Manmohan" sort="Parida, Manmohan" uniqKey="Parida M" first="Manmohan" last="Parida">Manmohan Parida</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Minekawa, Harumi" sort="Minekawa, Harumi" uniqKey="Minekawa H" first="Harumi" last="Minekawa">Harumi Minekawa</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Notomi, Tsugunori" sort="Notomi, Tsugunori" uniqKey="Notomi T" first="Tsugunori" last="Notomi">Tsugunori Notomi</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hasebe, Futoshi" sort="Hasebe, Futoshi" uniqKey="Hasebe F" first="Futoshi" last="Hasebe">Futoshi Hasebe</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Morita, Kouichi" sort="Morita, Kouichi" uniqKey="Morita K" first="Kouichi" last="Morita">Kouichi Morita</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">15131154</idno><idno type="pmc">404656</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC404656</idno><idno type="RBID">PMC:404656</idno><idno type="doi">10.1128/JCM.42.5.1956-1961.2004</idno><date when="2004">2004</date><idno type="wicri:Area/Pmc/Corpus">000645</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000645</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome Coronavirus</title><author><name sortKey="Thai, Hong Thi Cam" sort="Thai, Hong Thi Cam" uniqKey="Thai H" first="Hong Thi Cam" last="Thai">Hong Thi Cam Thai</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Le, Mai Quynh" sort="Le, Mai Quynh" uniqKey="Le M" first="Mai Quynh" last="Le">Mai Quynh Le</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Vuong, Cuong Duc" sort="Vuong, Cuong Duc" uniqKey="Vuong C" first="Cuong Duc" last="Vuong">Cuong Duc Vuong</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Parida, Manmohan" sort="Parida, Manmohan" uniqKey="Parida M" first="Manmohan" last="Parida">Manmohan Parida</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Minekawa, Harumi" sort="Minekawa, Harumi" uniqKey="Minekawa H" first="Harumi" last="Minekawa">Harumi Minekawa</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Notomi, Tsugunori" sort="Notomi, Tsugunori" uniqKey="Notomi T" first="Tsugunori" last="Notomi">Tsugunori Notomi</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hasebe, Futoshi" sort="Hasebe, Futoshi" uniqKey="Hasebe F" first="Futoshi" last="Hasebe">Futoshi Hasebe</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Morita, Kouichi" sort="Morita, Kouichi" uniqKey="Morita K" first="Kouichi" last="Morita">Kouichi Morita</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">Journal of Clinical Microbiology</title><idno type="ISSN">0095-1137</idno><idno type="eISSN">1098-660X</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The development and evaluation of a one-step single-tube accelerated real-time quantitative reverse transcription (RT) loop-mediated isothermal amplification (LAMP) assay is reported for rapid detection of the severe acute respiratory syndrome coronavirus (SARS-CoV) replicase gene. A total of 49 samples (15 throat washes, 13 throat swabs, and 21 combined throat and nasal swabs) collected from patients admitted to the Hanoi-French and Ninhbinh hospitals in Vietnam during the SARS epidemic were evaluated and compared to conventional RT-PCR. The RT-LAMP assay demonstrated 100-fold-greater sensitivity, with a detection limit of 0.01 PFU. The sensitivity and specificity of RT-LAMP assay for detecting viral RNA in clinical specimens with regard to RT-PCR were 100 and 87%, respectively. The specificity of the RT-LAMP assay was further validated by restriction analysis as well as nucleotide sequencing of the amplified product. The concentration of virus in most of the clinical samples was 0.1 PFU (0.1 to 10<sup>2</sup> PFU), as determined from the standard curve of SARS RT-LAMP and based on the time of positivity. The assay procedure is quite simple, wherein the amplification is carried out in a single tube under isothermal conditions at 63°C, and the result can be obtained in less than 1 h (as early as 11 min). Thus, the RT-LAMP assay reported here has the advantages of rapid amplification, simple operation, and easy detection and will be useful for rapid and reliable clinical diagnosis of SARS-CoV in developing countries.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Clin Microbiol</journal-id><journal-id journal-id-type="publisher-id">jcm</journal-id><journal-title>Journal of Clinical Microbiology</journal-title><issn pub-type="ppub">0095-1137</issn><issn pub-type="epub">1098-660X</issn><publisher><publisher-name>American Society for Microbiology</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">15131154</article-id><article-id pub-id-type="pmc">404656</article-id><article-id pub-id-type="publisher-id">1906-03</article-id><article-id pub-id-type="doi">10.1128/JCM.42.5.1956-1961.2004</article-id><article-categories><subj-group subj-group-type="heading"><subject>Virology</subject></subj-group></article-categories><title-group><article-title>Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome Coronavirus</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Thai</surname><given-names>Hong Thi Cam</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Le</surname><given-names>Mai Quynh</given-names></name><xref ref-type="aff" rid="aff1">2</xref></contrib><contrib contrib-type="author"><name><surname>Vuong</surname><given-names>Cuong Duc</given-names></name><xref ref-type="aff" rid="aff1">2</xref></contrib><contrib contrib-type="author"><name><surname>Parida</surname><given-names>Manmohan</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Minekawa</surname><given-names>Harumi</given-names></name><xref ref-type="aff" rid="aff1">3</xref></contrib><contrib contrib-type="author"><name><surname>Notomi</surname><given-names>Tsugunori</given-names></name><xref ref-type="aff" rid="aff1">3</xref></contrib><contrib contrib-type="author"><name><surname>Hasebe</surname><given-names>Futoshi</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Morita</surname><given-names>Kouichi</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="aff1">Department of Virology, Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8523,<label>1</label> Eiken Chemical Co. Ltd., Ohtawara, Tochigi 324-0036, Japan,<label>3</label> Department of Virology, National Institute of Hygiene and Epidemiology, Hanoi, Vietnam<label>2</label></aff><author-notes><fn id="cor1"><label>*</label><p>Corresponding author. Mailing address: Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan. Phone: 81 95 849 7829. Fax: 81 95 849 7830. E-mail: <email>moritak@net.nagasaki-u.ac.jp</email>.</p></fn></author-notes><pub-date pub-type="ppub"><month>5</month><year>2004</year></pub-date><volume>42</volume><issue>5</issue><fpage>1956</fpage><lpage>1961</lpage><history><date date-type="received"><day>7</day><month>11</month><year>2003</year></date><date date-type="rev-recd"><day>27</day><month>1</month><year>2003</year></date><date date-type="accepted"><day>2</day><month>2</month><year>2004</year></date></history><copyright-statement>Copyright © 2004, American Society for Microbiology</copyright-statement><copyright-year>2004</copyright-year><abstract><p>The development and evaluation of a one-step single-tube accelerated real-time quantitative reverse transcription (RT) loop-mediated isothermal amplification (LAMP) assay is reported for rapid detection of the severe acute respiratory syndrome coronavirus (SARS-CoV) replicase gene. A total of 49 samples (15 throat washes, 13 throat swabs, and 21 combined throat and nasal swabs) collected from patients admitted to the Hanoi-French and Ninhbinh hospitals in Vietnam during the SARS epidemic were evaluated and compared to conventional RT-PCR. The RT-LAMP assay demonstrated 100-fold-greater sensitivity, with a detection limit of 0.01 PFU. The sensitivity and specificity of RT-LAMP assay for detecting viral RNA in clinical specimens with regard to RT-PCR were 100 and 87%, respectively. The specificity of the RT-LAMP assay was further validated by restriction analysis as well as nucleotide sequencing of the amplified product. The concentration of virus in most of the clinical samples was 0.1 PFU (0.1 to 10<sup>2</sup> PFU), as determined from the standard curve of SARS RT-LAMP and based on the time of positivity. The assay procedure is quite simple, wherein the amplification is carried out in a single tube under isothermal conditions at 63°C, and the result can be obtained in less than 1 h (as early as 11 min). Thus, the RT-LAMP assay reported here has the advantages of rapid amplification, simple operation, and easy detection and will be useful for rapid and reliable clinical diagnosis of SARS-CoV in developing countries.</p></abstract></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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