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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Influenza A Virus Isolation, Culture and Identification</title>
<author><name sortKey="Eisfeld, Amie J" sort="Eisfeld, Amie J" uniqKey="Eisfeld A" first="Amie J." last="Eisfeld">Amie J. Eisfeld</name>
<affiliation><nlm:aff id="A1">Influenza Research Institute, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 575 Science Drive, Madison, WI 53711, USA</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Neumann, Gabriele" sort="Neumann, Gabriele" uniqKey="Neumann G" first="Gabriele" last="Neumann">Gabriele Neumann</name>
<affiliation><nlm:aff id="A1">Influenza Research Institute, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 575 Science Drive, Madison, WI 53711, USA</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name>
<affiliation><nlm:aff id="A1">Influenza Research Institute, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 575 Science Drive, Madison, WI 53711, USA</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="A2">ERATO Infection-Induced Host Responses Project, Japan Science and Technology Agency, Saitama 332-0012, Japan</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="A3">Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="A4">Department of Special Pathogens, International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan</nlm:aff>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">25321410</idno>
<idno type="pmc">5619698</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5619698</idno>
<idno type="RBID">PMC:5619698</idno>
<idno type="doi">10.1038/nprot.2014.180</idno>
<date when="2014">2014</date>
<idno type="wicri:Area/Pmc/Corpus">000817</idno>
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</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Influenza A Virus Isolation, Culture and Identification</title>
<author><name sortKey="Eisfeld, Amie J" sort="Eisfeld, Amie J" uniqKey="Eisfeld A" first="Amie J." last="Eisfeld">Amie J. Eisfeld</name>
<affiliation><nlm:aff id="A1">Influenza Research Institute, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 575 Science Drive, Madison, WI 53711, USA</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Neumann, Gabriele" sort="Neumann, Gabriele" uniqKey="Neumann G" first="Gabriele" last="Neumann">Gabriele Neumann</name>
<affiliation><nlm:aff id="A1">Influenza Research Institute, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 575 Science Drive, Madison, WI 53711, USA</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Kawaoka, Yoshihiro" sort="Kawaoka, Yoshihiro" uniqKey="Kawaoka Y" first="Yoshihiro" last="Kawaoka">Yoshihiro Kawaoka</name>
<affiliation><nlm:aff id="A1">Influenza Research Institute, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 575 Science Drive, Madison, WI 53711, USA</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="A2">ERATO Infection-Induced Host Responses Project, Japan Science and Technology Agency, Saitama 332-0012, Japan</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="A3">Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="A4">Department of Special Pathogens, International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan</nlm:aff>
</affiliation>
</author>
</analytic>
<series><title level="j">Nature protocols</title>
<idno type="ISSN">1754-2189</idno>
<idno type="eISSN">1750-2799</idno>
<imprint><date when="2014">2014</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><title>SUMMARY</title>
<p id="P1">Influenza A viruses (IAV) cause epidemics and pandemics that result in considerable financial burden and loss of human life. To manage annual IAV epidemics and prepare for future pandemics, improved understanding of how IAVs emerge, transmit, cause disease, and acquire pandemic potential is urgently needed. Fundamental techniques essential for procuring such knowledge are IAV isolation and culture from experimental and surveillance samples. Here, we present a detailed protocol for IAV sample collection and processing, amplification in chicken eggs and mammalian cells, and identification from samples containing unknown pathogens. This protocol is robust, and allows for generation of virus cultures that can be used for downstream analyses. Once experimental or surveillance samples are obtained, virus cultures can be generated and the presence of IAV can be verified in 3–5 days. Increased time-frames may be required for less experienced laboratory personnel, or when large numbers of samples will be processed.</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>
<pmc-dir>properties manuscript</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-journal-id">101284307</journal-id>
<journal-id journal-id-type="pubmed-jr-id">33047</journal-id>
<journal-id journal-id-type="nlm-ta">Nat Protoc</journal-id>
<journal-id journal-id-type="iso-abbrev">Nat Protoc</journal-id>
<journal-title-group><journal-title>Nature protocols</journal-title>
</journal-title-group>
<issn pub-type="ppub">1754-2189</issn>
<issn pub-type="epub">1750-2799</issn>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">25321410</article-id>
<article-id pub-id-type="pmc">5619698</article-id>
<article-id pub-id-type="doi">10.1038/nprot.2014.180</article-id>
<article-id pub-id-type="manuscript">NIHMS902603</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Influenza A Virus Isolation, Culture and Identification</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Eisfeld</surname>
<given-names>Amie J.</given-names>
</name>
<pmc-comment>aefenney@vetmed.wisc.edu </pmc-comment>
<xref ref-type="aff" rid="A1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Neumann</surname>
<given-names>Gabriele</given-names>
</name>
<pmc-comment>neumanng@vetmed.wisc.edu </pmc-comment>
<xref ref-type="aff" rid="A1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Kawaoka</surname>
<given-names>Yoshihiro</given-names>
</name>
<xref ref-type="aff" rid="A1">1</xref>
<xref ref-type="aff" rid="A2">2</xref>
<xref ref-type="aff" rid="A3">3</xref>
<xref ref-type="aff" rid="A4">4</xref>
<xref rid="FN1" ref-type="author-notes">*</xref>
</contrib>
</contrib-group>
<aff id="A1"><label>1</label>
Influenza Research Institute, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 575 Science Drive, Madison, WI 53711, USA</aff>
<aff id="A2"><label>2</label>
ERATO Infection-Induced Host Responses Project, Japan Science and Technology Agency, Saitama 332-0012, Japan</aff>
<aff id="A3"><label>3</label>
Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan</aff>
<aff id="A4"><label>4</label>
Department of Special Pathogens, International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan</aff>
<author-notes><corresp id="FN1"><label>*</label>
<bold>Corresponding author’s telephone and fax numbers:</bold>
+1 (608) 265-4925 (office); +1 (608) 262-9641 (fax), <email>kawaokay@svm.vetmed.wisc.edu</email>
</corresp>
<fn fn-type="COI-statement" id="FN3"><p><bold>COMPETING FINANCIAL INTERESTS</bold>
</p>
<p>The authors declare that they have no competing financial interests.</p>
</fn>
</author-notes>
<pub-date pub-type="nihms-submitted"><day>1</day>
<month>9</month>
<year>2017</year>
</pub-date>
<pub-date pub-type="epub"><day>16</day>
<month>10</month>
<year>2014</year>
</pub-date>
<pub-date pub-type="ppub"><month>11</month>
<year>2014</year>
</pub-date>
<pub-date pub-type="pmc-release"><day>28</day>
<month>9</month>
<year>2017</year>
</pub-date>
<volume>9</volume>
<issue>11</issue>
<fpage>2663</fpage>
<lpage>2681</lpage>
<pmc-comment>elocation-id from pubmed: 10.1038/nprot.2014.180</pmc-comment>
<abstract><title>SUMMARY</title>
<p id="P1">Influenza A viruses (IAV) cause epidemics and pandemics that result in considerable financial burden and loss of human life. To manage annual IAV epidemics and prepare for future pandemics, improved understanding of how IAVs emerge, transmit, cause disease, and acquire pandemic potential is urgently needed. Fundamental techniques essential for procuring such knowledge are IAV isolation and culture from experimental and surveillance samples. Here, we present a detailed protocol for IAV sample collection and processing, amplification in chicken eggs and mammalian cells, and identification from samples containing unknown pathogens. This protocol is robust, and allows for generation of virus cultures that can be used for downstream analyses. Once experimental or surveillance samples are obtained, virus cultures can be generated and the presence of IAV can be verified in 3–5 days. Increased time-frames may be required for less experienced laboratory personnel, or when large numbers of samples will be processed.</p>
</abstract>
</article-meta>
</front>
</pmc>
</record>
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