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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Multiplexed Luminex xMAP Assay for Detection and Identification of Five Adenovirus Serotypes Associated with Epidemics of Respiratory Disease in Adults<xref ref-type="fn" rid="fn2">▿</xref> <xref ref-type="fn" rid="fn1">†</xref> </title><author><name sortKey="Washington, Cicely" sort="Washington, Cicely" uniqKey="Washington C" first="Cicely" last="Washington">Cicely Washington</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Metzgar, David" sort="Metzgar, David" uniqKey="Metzgar D" first="David" last="Metzgar">David Metzgar</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hazb N, Manzour Hernando" sort="Hazb N, Manzour Hernando" uniqKey="Hazb N M" first="Manzour Hernando" last="Hazb N">Manzour Hernando Hazb N</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Binn, Leonard" sort="Binn, Leonard" uniqKey="Binn L" first="Leonard" last="Binn">Leonard Binn</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Lyons, Arthur" sort="Lyons, Arthur" uniqKey="Lyons A" first="Arthur" last="Lyons">Arthur Lyons</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Coward, Carl" sort="Coward, Carl" uniqKey="Coward C" first="Carl" last="Coward">Carl Coward</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kuschner, Robert" sort="Kuschner, Robert" uniqKey="Kuschner R" first="Robert" last="Kuschner">Robert Kuschner</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">20410343</idno><idno type="pmc">2884482</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2884482</idno><idno type="RBID">PMC:2884482</idno><idno type="doi">10.1128/JCM.00029-10</idno><date when="2010">2010</date><idno type="wicri:Area/Pmc/Corpus">000445</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000445</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Multiplexed Luminex xMAP Assay for Detection and Identification of Five Adenovirus Serotypes Associated with Epidemics of Respiratory Disease in Adults<xref ref-type="fn" rid="fn2">▿</xref> <xref ref-type="fn" rid="fn1">†</xref> </title><author><name sortKey="Washington, Cicely" sort="Washington, Cicely" uniqKey="Washington C" first="Cicely" last="Washington">Cicely Washington</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Metzgar, David" sort="Metzgar, David" uniqKey="Metzgar D" first="David" last="Metzgar">David Metzgar</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hazb N, Manzour Hernando" sort="Hazb N, Manzour Hernando" uniqKey="Hazb N M" first="Manzour Hernando" last="Hazb N">Manzour Hernando Hazb N</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Binn, Leonard" sort="Binn, Leonard" uniqKey="Binn L" first="Leonard" last="Binn">Leonard Binn</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Lyons, Arthur" sort="Lyons, Arthur" uniqKey="Lyons A" first="Arthur" last="Lyons">Arthur Lyons</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Coward, Carl" sort="Coward, Carl" uniqKey="Coward C" first="Carl" last="Coward">Carl Coward</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kuschner, Robert" sort="Kuschner, Robert" uniqKey="Kuschner R" first="Robert" last="Kuschner">Robert Kuschner</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="2010">2010</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Several serotypes of human adenovirus (HAdV) cause acute respiratory disease (ARD) among healthy adults, sometimes generating broad outbreaks with high attack rates and occasional fatalities. Timely serotype identification provides valuable epidemiological information and significantly contributes to prevention (vaccination) strategies. The prevalence of specific serotypes causing ARD varies geographically. HAdV-3, HAdV-4, HAdV-7, HAdV-14, and HAdV-21 are the serotypes most commonly found in adult populations in the Western Hemisphere. Unfortunately, conventional serotype identification is a tedious process which can take a week or longer. For this reason, new molecular methods for serotype identification are needed. Commercially available rapid antigen and PCR assays for the detection of HAdV are universal but do not distinguish between the different serotypes. We describe the development of a sensitive and specific multiplex assay capable of identifying serotypes 3, 4, 7, 14, and 21. Two sets of primers were used for nonspecific (universal) PCR amplification, and serotype-specific probes coupled to Luminex tags were used for target-specific extension (TSE). PCR and TSE primers were designed using known hexon gene sequences of HAdV. The TSE products of HAdV-3, HAdV-4, HAdV-7, HAdV-14, and HAdV-21 were correctly identified using the Luminex xMAP fluid microsphere-based array system. No cross-reactivity with other respiratory pathogens or other HAdV serotypes was observed. This multiplexed assay can be expanded to include more serotypes and will allow broad and rapid detection and identification of adenoviral serotypes in a high-throughput environment.</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-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 (ASM)</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">20410343</article-id><article-id pub-id-type="pmc">2884482</article-id><article-id pub-id-type="publisher-id">0029-10</article-id><article-id pub-id-type="doi">10.1128/JCM.00029-10</article-id><article-categories><subj-group subj-group-type="heading"><subject>Virology</subject></subj-group></article-categories><title-group><article-title>Multiplexed Luminex xMAP Assay for Detection and Identification of Five Adenovirus Serotypes Associated with Epidemics of Respiratory Disease in Adults<xref ref-type="fn" rid="fn2">▿</xref> <xref ref-type="fn" rid="fn1">†</xref> </article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Washington</surname><given-names>Cicely</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib><contrib contrib-type="author"><name><surname>Metzgar</surname><given-names>David</given-names></name><xref ref-type="aff" rid="aff1">2</xref></contrib><contrib contrib-type="author"><name><surname>Hazbón</surname><given-names>Manzour Hernando</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Binn</surname><given-names>Leonard</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Lyons</surname><given-names>Arthur</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Coward</surname><given-names>Carl</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Kuschner</surname><given-names>Robert</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib></contrib-group><aff id="aff1">Division of Viral Diseases, Walter Reed Army Institute of Research, Silver Spring, Maryland 20910,<label>1</label> Department of Respiratory Diseases Research, Naval Health Research Center, San Diego, California 92152<label>2</label></aff><author-notes><corresp id="cor1"><label>*</label>Corresponding author. Mailing address: 503 Robert Grant Ave., Silver Spring, MD 20910. Phone: (301) 319-9824. Fax: (301) 319-9661. E-mail: <email>Cicely.Washington@us.army.mil</email></corresp></author-notes><pub-date pub-type="ppub"><month>6</month><year>2010</year></pub-date><pub-date pub-type="epub"><day>21</day><month>4</month><year>2010</year></pub-date><volume>48</volume><issue>6</issue><fpage>2217</fpage><lpage>2222</lpage><history><date date-type="received"><day>6</day><month>1</month><year>2010</year></date><date date-type="rev-recd"><day>19</day><month>2</month><year>2010</year></date><date date-type="accepted"><day>13</day><month>4</month><year>2010</year></date></history><permissions><copyright-statement>Copyright © 2010, American Society for Microbiology</copyright-statement><copyright-year>2010</copyright-year></permissions><self-uri xlink:title="pdf" xlink:href="zjm00610002217.pdf"></self-uri><abstract><p>Several serotypes of human adenovirus (HAdV) cause acute respiratory disease (ARD) among healthy adults, sometimes generating broad outbreaks with high attack rates and occasional fatalities. Timely serotype identification provides valuable epidemiological information and significantly contributes to prevention (vaccination) strategies. The prevalence of specific serotypes causing ARD varies geographically. HAdV-3, HAdV-4, HAdV-7, HAdV-14, and HAdV-21 are the serotypes most commonly found in adult populations in the Western Hemisphere. Unfortunately, conventional serotype identification is a tedious process which can take a week or longer. For this reason, new molecular methods for serotype identification are needed. Commercially available rapid antigen and PCR assays for the detection of HAdV are universal but do not distinguish between the different serotypes. We describe the development of a sensitive and specific multiplex assay capable of identifying serotypes 3, 4, 7, 14, and 21. Two sets of primers were used for nonspecific (universal) PCR amplification, and serotype-specific probes coupled to Luminex tags were used for target-specific extension (TSE). PCR and TSE primers were designed using known hexon gene sequences of HAdV. The TSE products of HAdV-3, HAdV-4, HAdV-7, HAdV-14, and HAdV-21 were correctly identified using the Luminex xMAP fluid microsphere-based array system. No cross-reactivity with other respiratory pathogens or other HAdV serotypes was observed. This multiplexed assay can be expanded to include more serotypes and will allow broad and rapid detection and identification of adenoviral serotypes in a high-throughput environment.</p></abstract></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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