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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Sensitive and Rapid Detection of the New Delhi Metallo-Beta-Lactamase Gene by Loop-Mediated Isothermal Amplification</title><author><name sortKey="Liu, Wei" sort="Liu, Wei" uniqKey="Liu W" first="Wei" last="Liu">Wei Liu</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Zou, Dayang" sort="Zou, Dayang" uniqKey="Zou D" first="Dayang" last="Zou">Dayang Zou</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Li, Yan" sort="Li, Yan" uniqKey="Li Y" first="Yan" last="Li">Yan Li</name><affiliation><nlm:aff id="aff2">Department of Respiratory Diseases, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Wang, Xuesong" sort="Wang, Xuesong" uniqKey="Wang X" first="Xuesong" last="Wang">Xuesong Wang</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="He, Xiang" sort="He, Xiang" uniqKey="He X" first="Xiang" last="He">Xiang He</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Wei, Xiao" sort="Wei, Xiao" uniqKey="Wei X" first="Xiao" last="Wei">Xiao Wei</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Shao, Changlin" sort="Shao, Changlin" uniqKey="Shao C" first="Changlin" last="Shao">Changlin Shao</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Li, Xuelian" sort="Li, Xuelian" uniqKey="Li X" first="Xuelian" last="Li">Xuelian Li</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Shang, Wei" sort="Shang, Wei" uniqKey="Shang W" first="Wei" last="Shang">Wei Shang</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Yu, Kaitao" sort="Yu, Kaitao" uniqKey="Yu K" first="Kaitao" last="Yu">Kaitao Yu</name><affiliation><nlm:aff id="aff2">Department of Respiratory Diseases, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Liu, Dawei" sort="Liu, Dawei" uniqKey="Liu D" first="Dawei" last="Liu">Dawei Liu</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Li, Yunmei" sort="Li, Yunmei" uniqKey="Li Y" first="Yunmei" last="Li">Yunmei Li</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Guo, Jing" sort="Guo, Jing" uniqKey="Guo J" first="Jing" last="Guo">Jing Guo</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Yin, Zhitao" sort="Yin, Zhitao" uniqKey="Yin Z" first="Zhitao" last="Yin">Zhitao Yin</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Yuan, Jing" sort="Yuan, Jing" uniqKey="Yuan J" first="Jing" last="Yuan">Jing Yuan</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">22357496</idno><idno type="pmc">3347096</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3347096</idno><idno type="RBID">PMC:3347096</idno><idno type="doi">10.1128/JCM.06647-11</idno><date when="2012">2012</date><idno type="wicri:Area/Pmc/Corpus">000444</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000444</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Sensitive and Rapid Detection of the New Delhi Metallo-Beta-Lactamase Gene by Loop-Mediated Isothermal Amplification</title><author><name sortKey="Liu, Wei" sort="Liu, Wei" uniqKey="Liu W" first="Wei" last="Liu">Wei Liu</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Zou, Dayang" sort="Zou, Dayang" uniqKey="Zou D" first="Dayang" last="Zou">Dayang Zou</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Li, Yan" sort="Li, Yan" uniqKey="Li Y" first="Yan" last="Li">Yan Li</name><affiliation><nlm:aff id="aff2">Department of Respiratory Diseases, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Wang, Xuesong" sort="Wang, Xuesong" uniqKey="Wang X" first="Xuesong" last="Wang">Xuesong Wang</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="He, Xiang" sort="He, Xiang" uniqKey="He X" first="Xiang" last="He">Xiang He</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Wei, Xiao" sort="Wei, Xiao" uniqKey="Wei X" first="Xiao" last="Wei">Xiao Wei</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Shao, Changlin" sort="Shao, Changlin" uniqKey="Shao C" first="Changlin" last="Shao">Changlin Shao</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Li, Xuelian" sort="Li, Xuelian" uniqKey="Li X" first="Xuelian" last="Li">Xuelian Li</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Shang, Wei" sort="Shang, Wei" uniqKey="Shang W" first="Wei" last="Shang">Wei Shang</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Yu, Kaitao" sort="Yu, Kaitao" uniqKey="Yu K" first="Kaitao" last="Yu">Kaitao Yu</name><affiliation><nlm:aff id="aff2">Department of Respiratory Diseases, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Liu, Dawei" sort="Liu, Dawei" uniqKey="Liu D" first="Dawei" last="Liu">Dawei Liu</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Li, Yunmei" sort="Li, Yunmei" uniqKey="Li Y" first="Yunmei" last="Li">Yunmei Li</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Guo, Jing" sort="Guo, Jing" uniqKey="Guo J" first="Jing" last="Guo">Jing Guo</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Yin, Zhitao" sort="Yin, Zhitao" uniqKey="Yin Z" first="Zhitao" last="Yin">Zhitao Yin</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</nlm:aff></affiliation></author><author><name sortKey="Yuan, Jing" sort="Yuan, Jing" uniqKey="Yuan J" first="Jing" last="Yuan">Jing Yuan</name><affiliation><nlm:aff id="aff1">Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</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="2012">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>New Delhi metallo-β-lactamase 1 (NDM-1), which is associated with resistance to carbapenem, was first reported in 2008. A sensitive and rapid molecular assay to detect the plasmid <italic>bla</italic><sub>NDM-1</sub> in clinical isolates is needed to control its spread. We describe a loop-mediated isothermal amplification (LAMP) assay for the rapid detection of <italic>bla</italic><sub>NDM-1</sub> from pure culture and sputum, urine, and fecal samples. Eight sets of primers were designed to recognize six or eight distinct sequences on target <italic>bla</italic><sub>NDM-1</sub>, and one set was selected as the most appropriate set of primers for its rapid detection. The specificity and sensitivity of the primers in the LAMP reactions for <italic>bla</italic><sub>NDM-1</sub> detection were determined. The sensitivity of the LAMP assay for <italic>bla</italic><sub>NDM-1</sub> detection in sputum, urine, and fecal samples was also tested. Two methods, namely, monitoring of turbidity and addition of calcein to the reaction tube, were used to determine negative and positive results. The results showed that target DNA was amplified and visualized by the two detection methods within 70 min at an isothermal temperature of 65°C. The sensitivity of LAMP, with a detection limit of 10.70 pg/μl DNA, was 100-fold greater than that of PCR. Thirteen infection bacterial strains without <italic>bla</italic><sub>NDM-1</sub> were selected for testing of specificity, and the results of the amplification were negative, which showed that the primers had good levels of specificity. The LAMP method reported here is demonstrated to be a potentially valuable means for the detection of <italic>bla</italic><sub>NDM-1</sub> and rapid clinical diagnosis, being fast, simple, and low in cost.</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="iso-abbrev">J. Clin. Microbiol</journal-id><journal-id journal-id-type="hwp">jcm</journal-id><journal-id journal-id-type="pmc">jcm</journal-id><journal-id journal-id-type="publisher-id">JCM</journal-id><journal-title-group><journal-title>Journal of Clinical Microbiology</journal-title></journal-title-group><issn pub-type="ppub">0095-1137</issn><issn pub-type="epub">1098-660X</issn><publisher><publisher-name>American Society for Microbiology</publisher-name><publisher-loc>1752 N St., N.W., Washington, DC</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">22357496</article-id><article-id pub-id-type="pmc">3347096</article-id><article-id pub-id-type="publisher-id">06647-11</article-id><article-id pub-id-type="doi">10.1128/JCM.06647-11</article-id><article-categories><subj-group subj-group-type="heading"><subject>Bacteriology</subject></subj-group></article-categories><title-group><article-title>Sensitive and Rapid Detection of the New Delhi Metallo-Beta-Lactamase Gene by Loop-Mediated Isothermal Amplification</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Liu</surname><given-names>Wei</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Zou</surname><given-names>Dayang</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Yan</given-names></name><xref ref-type="aff" rid="aff2"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Xuesong</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>He</surname><given-names>Xiang</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Wei</surname><given-names>Xiao</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Shao</surname><given-names>Changlin</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Xuelian</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Shang</surname><given-names>Wei</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Yu</surname><given-names>Kaitao</given-names></name><xref ref-type="aff" rid="aff2"><sup>b</sup></xref></contrib><contrib contrib-type="author"><name><surname>Liu</surname><given-names>Dawei</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Yunmei</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Guo</surname><given-names>Jing</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Yin</surname><given-names>Zhitao</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Yuan</surname><given-names>Jing</given-names></name><xref ref-type="aff" rid="aff1"><sup>a</sup></xref></contrib><aff id="aff1"><label>a</label>Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China</aff><aff id="aff2"><label>b</label>Department of Respiratory Diseases, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, China</aff></contrib-group><author-notes><corresp>Address correspondence to Jing Yuan, <email>yuanjing6216@163.com</email>.</corresp><fn fn-type="equal"><p>W.L., D.Z., and Y.L. contributed equally to this article.</p></fn></author-notes><pub-date pub-type="ppub"><month>5</month><year>2012</year></pub-date><volume>50</volume><issue>5</issue><fpage>1580</fpage><lpage>1585</lpage><history><date date-type="received"><day>15</day><month>12</month><year>2011</year></date><date date-type="rev-request"><day>10</day><month>1</month><year>2012</year></date><date date-type="accepted"><day>10</day><month>2</month><year>2012</year></date></history><permissions><copyright-statement>Copyright © 2012, American Society for Microbiology. All Rights Reserved.</copyright-statement><copyright-year>2012</copyright-year><copyright-holder>American Society for Microbiology</copyright-holder></permissions><self-uri xlink:title="pdf" xlink:type="simple" xlink:href="zjm00512001580.pdf"></self-uri><abstract><p>New Delhi metallo-β-lactamase 1 (NDM-1), which is associated with resistance to carbapenem, was first reported in 2008. A sensitive and rapid molecular assay to detect the plasmid <italic>bla</italic><sub>NDM-1</sub> in clinical isolates is needed to control its spread. We describe a loop-mediated isothermal amplification (LAMP) assay for the rapid detection of <italic>bla</italic><sub>NDM-1</sub> from pure culture and sputum, urine, and fecal samples. Eight sets of primers were designed to recognize six or eight distinct sequences on target <italic>bla</italic><sub>NDM-1</sub>, and one set was selected as the most appropriate set of primers for its rapid detection. The specificity and sensitivity of the primers in the LAMP reactions for <italic>bla</italic><sub>NDM-1</sub> detection were determined. The sensitivity of the LAMP assay for <italic>bla</italic><sub>NDM-1</sub> detection in sputum, urine, and fecal samples was also tested. Two methods, namely, monitoring of turbidity and addition of calcein to the reaction tube, were used to determine negative and positive results. The results showed that target DNA was amplified and visualized by the two detection methods within 70 min at an isothermal temperature of 65°C. The sensitivity of LAMP, with a detection limit of 10.70 pg/μl DNA, was 100-fold greater than that of PCR. Thirteen infection bacterial strains without <italic>bla</italic><sub>NDM-1</sub> were selected for testing of specificity, and the results of the amplification were negative, which showed that the primers had good levels of specificity. The LAMP method reported here is demonstrated to be a potentially valuable means for the detection of <italic>bla</italic><sub>NDM-1</sub> and rapid clinical diagnosis, being fast, simple, and low in cost.</p></abstract></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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