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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Combined PCR-Heteroduplex Mobility Assay for Detection and Differentiation of Influenza A Viruses from Different Animal Species</title><author><name sortKey="Ellis, Joanna S" sort="Ellis, Joanna S" uniqKey="Ellis J" first="Joanna S." last="Ellis">Joanna S. Ellis</name></author><author><name sortKey="Zambon, Maria C" sort="Zambon, Maria C" uniqKey="Zambon M" first="Maria C." last="Zambon">Maria C. Zambon</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">11682536</idno><idno type="pmc">88493</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC88493</idno><idno type="RBID">PMC:88493</idno><idno type="doi">10.1128/JCM.39.11.4097-4102.2001</idno><date when="2001">2001</date><idno type="wicri:Area/Pmc/Corpus">000131</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000131</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Combined PCR-Heteroduplex Mobility Assay for Detection and Differentiation of Influenza A Viruses from Different Animal Species</title><author><name sortKey="Ellis, Joanna S" sort="Ellis, Joanna S" uniqKey="Ellis J" first="Joanna S." last="Ellis">Joanna S. Ellis</name></author><author><name sortKey="Zambon, Maria C" sort="Zambon, Maria C" uniqKey="Zambon M" first="Maria C." last="Zambon">Maria C. Zambon</name></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="2001">2001</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Transfer of influenza A viruses from animal hosts to man may lead to the emergence of new human pandemic strains. The early detection and identification of such events are therefore paramount in the surveillance of influenza viruses. To detect and partially characterize influenza A viruses from different animal species, a combined reverse transcription (RT)-PCR heteroduplex mobility assay (HMA) was designed. This M gene RT-PCR was shown to be sensitive and specific for the detection of human, avian, and swine influenza A viruses. PCR amplicons from human, avian, and swine viruses of 15 different subtypes, with between 1.9 and 21.4% nucleotide divergence, were differentiated by HMA. Sequencing of the amplicons showed that the heteroduplex mobility patterns correlated with the sequence divergence between test and reference DNA. The application of the RT-PCR HMA method for rapid screening of samples was assessed with a reference panel of viruses of human, avian, and swine origin. The avian H9N2 virus A/HongKong/1073/99, which crossed the species barrier to humans, was screened against the reference panel. It was found to be most closely related to the avian A/Quail/HongKong/G1/97 H9N2 reference PCR product. Sequence analysis showed a nucleotide divergence of 1.1% between the A/Quail/HongKong/G1/97 and A/HongKong/1073/99 amplicons. From the results of our work, we consider the RT-PCR HMA method described to offer a rapid and sensitive means for screening for novel or unusual influenza viruses.</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">J CLIN MICROBIOL</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">11682536</article-id><article-id pub-id-type="pmc">88493</article-id><article-id pub-id-type="publisher-id">1636</article-id><article-id pub-id-type="doi">10.1128/JCM.39.11.4097-4102.2001</article-id><article-categories><subj-group subj-group-type="heading"><subject>Virology</subject></subj-group></article-categories><title-group><article-title>Combined PCR-Heteroduplex Mobility Assay for Detection and Differentiation of Influenza A Viruses from Different Animal Species</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Ellis</surname><given-names>Joanna S.</given-names></name><xref ref-type="author-notes" rid="FN150">*</xref></contrib><contrib contrib-type="author"><name><surname>Zambon</surname><given-names>Maria C.</given-names></name></contrib></contrib-group><aff id="N0x9aae610.0xa4ea918">Respiratory Virus Unit, Enteric, Respiratory and Neurological Virus Laboratory, Public Health Laboratory Service, Central Public Health Laboratory, Colindale, London NW9 5HT, United Kingdom</aff><author-notes><fn id="FN150"><label>*</label><p>Corresponding author. Mailing address: Respiratory Virus Unit, Enteric, Respiratory and Neurological Virus Lab., Public Health Lab. Service, Central Public Health Laboratory, 61 Colindale Ave., Colindale, London NW9 5HT, United Kingdom. Phone: 020 8200 4400. Fax: 020 8200 1569. E-mail: <email>jellis@phls.nhs.uk</email>.</p></fn></author-notes><pub-date pub-type="ppub"><month>11</month><year>2001</year></pub-date><volume>39</volume><issue>11</issue><fpage>4097</fpage><lpage>4102</lpage><history><date date-type="received"><day>19</day><month>12</month><year>2000</year></date><date date-type="rev-request"><day>13</day><month>1</month><year>2001</year></date><date date-type="accepted"><day>8</day><month>9</month><year>2001</year></date></history><copyright-statement>Copyright © 2001, American Society for Microbiology</copyright-statement><copyright-year>2001</copyright-year><abstract><p>Transfer of influenza A viruses from animal hosts to man may lead to the emergence of new human pandemic strains. The early detection and identification of such events are therefore paramount in the surveillance of influenza viruses. To detect and partially characterize influenza A viruses from different animal species, a combined reverse transcription (RT)-PCR heteroduplex mobility assay (HMA) was designed. This M gene RT-PCR was shown to be sensitive and specific for the detection of human, avian, and swine influenza A viruses. PCR amplicons from human, avian, and swine viruses of 15 different subtypes, with between 1.9 and 21.4% nucleotide divergence, were differentiated by HMA. Sequencing of the amplicons showed that the heteroduplex mobility patterns correlated with the sequence divergence between test and reference DNA. The application of the RT-PCR HMA method for rapid screening of samples was assessed with a reference panel of viruses of human, avian, and swine origin. The avian H9N2 virus A/HongKong/1073/99, which crossed the species barrier to humans, was screened against the reference panel. It was found to be most closely related to the avian A/Quail/HongKong/G1/97 H9N2 reference PCR product. Sequence analysis showed a nucleotide divergence of 1.1% between the A/Quail/HongKong/G1/97 and A/HongKong/1073/99 amplicons. From the results of our work, we consider the RT-PCR HMA method described to offer a rapid and sensitive means for screening for novel or unusual influenza viruses.</p></abstract></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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