Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses
Identifieur interne : 001807 ( Istex/Corpus ); précédent : 001806; suivant : 001808Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses
Auteurs : Ilona Stefa Ska ; Tomasz Dzieciatkowski ; Lidia B. Brydak ; Magdalena RomanowskaSource :
- Archives of Virology [ 0304-8608 ] ; 2013.
Abstract
Abstract: This study was performed to develop real-time PCR (qPCR) for detection of human seasonal and avian influenza viruses in duplex format. First duplex qPCR detects haemagglutinin (HA) gene of influenza virus A(H1N1)pdm09 and HA gene of influenza virus A(H3N2), the second reaction detects neuraminidase (NA) gene of influenza virus A(H3N2) and NA gene of influenza virus A(H1N1)pdm09 and A(H5N1), and the third reaction detects HA gene of influenza A(H5N1) and nonstructural protein gene of influenza B virus. Primers and probes were designed using multiple alignments of target gene sequences of different reference strains. Assays were optimised for identical thermocycling conditions. Their specificity was confirmed by conventional PCR and monoplex qPCR with nucleic acids isolated from different influenza viruses and other respiratory pathogens. Plasmid constructs with a fragment of specific gene were used to assess sensitivity of the assay. The limit of detection ranged from 27 to 96 cDNA copies/reaction. Clinical specimens (n = 107) have been tested using new assays, immunofluorescence and monoplex qRT-PCR. It has been shown that developed assays have been capable of rapid and accurate simultaneous detection and differentiation of influenza viruses. They are more sensitive than immunofluorescence and at least as sensitive as monoplex qRT-PCR.
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DOI: 10.1007/s00705-013-1648-0
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Brydak</name><affiliation><mods:affiliation>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw, Poland</mods:affiliation></affiliation><affiliation><mods:affiliation>Department of Immunology, Faculty of Biology, University of Szczecin, 3a Felczaka Street, 71-412, Szczecin, Poland</mods:affiliation></affiliation></author><author><name sortKey="Romanowska, Magdalena" sort="Romanowska, Magdalena" uniqKey="Romanowska M" first="Magdalena" last="Romanowska">Magdalena Romanowska</name><affiliation><mods:affiliation>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw, Poland</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Archives of Virology</title><title level="j" type="abbrev">Arch Virol</title><idno type="ISSN">0304-8608</idno><idno type="eISSN">1432-8798</idno><imprint><publisher ref="https://scientific-publisher.data.istex.fr/ark:/67375/H02-SWLMH5L1-1">Springer Vienna</publisher><pubPlace>Vienna</pubPlace><date type="published" when="2013">2013</date><biblScope unit="vol" from="158" to="158">158</biblScope><biblScope unit="issue" from="8" to="8">8</biblScope><biblScope unit="page" from="1743">1743</biblScope><biblScope unit="page" to="1753">1753</biblScope></imprint><idno type="ISSN">0304-8608</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0304-8608</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: This study was performed to develop real-time PCR (qPCR) for detection of human seasonal and avian influenza viruses in duplex format. First duplex qPCR detects haemagglutinin (HA) gene of influenza virus A(H1N1)pdm09 and HA gene of influenza virus A(H3N2), the second reaction detects neuraminidase (NA) gene of influenza virus A(H3N2) and NA gene of influenza virus A(H1N1)pdm09 and A(H5N1), and the third reaction detects HA gene of influenza A(H5N1) and nonstructural protein gene of influenza B virus. Primers and probes were designed using multiple alignments of target gene sequences of different reference strains. Assays were optimised for identical thermocycling conditions. Their specificity was confirmed by conventional PCR and monoplex qPCR with nucleic acids isolated from different influenza viruses and other respiratory pathogens. Plasmid constructs with a fragment of specific gene were used to assess sensitivity of the assay. The limit of detection ranged from 27 to 96 cDNA copies/reaction. Clinical specimens (n = 107) have been tested using new assays, immunofluorescence and monoplex qRT-PCR. It has been shown that developed assays have been capable of rapid and accurate simultaneous detection and differentiation of influenza viruses. They are more sensitive than immunofluorescence and at least as sensitive as monoplex qRT-PCR.</div></front></TEI><istex><corpusName>springer-journals</corpusName><author><json:item><name>Ilona Stefańska</name><affiliations><json:string>Department of Fermentation Technology, Institute of Agricultural and Food Biotechnology, 36 Rakowiecka Street, 02-532, Warsaw, Poland</json:string><json:string>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw, Poland</json:string><json:string>E-mail: i.stefanska@gmail.com</json:string></affiliations></json:item><json:item><name>Tomasz Dzieciatkowski</name><affiliations><json:string>Chair and Department of Medical Microbiology, Medical University of Warsaw, 5 Chalubinski Street, 02-004, Warsaw, Poland</json:string></affiliations></json:item><json:item><name>Lidia B. Brydak</name><affiliations><json:string>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw, Poland</json:string><json:string>Department of Immunology, Faculty of Biology, University of Szczecin, 3a Felczaka Street, 71-412, Szczecin, Poland</json:string></affiliations></json:item><json:item><name>Magdalena Romanowska</name><affiliations><json:string>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw, Poland</json:string></affiliations></json:item></author><articleId><json:string>1648</json:string><json:string>s00705-013-1648-0</json:string></articleId><arkIstex>ark:/67375/VQC-9JNV2659-Q</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>OriginalPaper</json:string></originalGenre><abstract>Abstract: This study was performed to develop real-time PCR (qPCR) for detection of human seasonal and avian influenza viruses in duplex format. 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TocLevels="0"><ArticleID>1648</ArticleID><ArticleDOI>10.1007/s00705-013-1648-0</ArticleDOI><ArticleSequenceNumber>11</ArticleSequenceNumber><ArticleTitle Language="En" OutputMedium="All">Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses</ArticleTitle><ArticleCategory>Original Article</ArticleCategory><ArticleFirstPage>1743</ArticleFirstPage><ArticleLastPage>1753</ArticleLastPage><ArticleHistory><RegistrationDate><Year>2013</Year><Month>2</Month><Day>19</Day></RegistrationDate><Received><Year>2012</Year><Month>10</Month><Day>22</Day></Received><Accepted><Year>2013</Year><Month>1</Month><Day>23</Day></Accepted><OnlineDate><Year>2013</Year><Month>3</Month><Day>21</Day></OnlineDate></ArticleHistory><ArticleCopyright><CopyrightHolderName>Springer-Verlag Wien</CopyrightHolderName><CopyrightYear>2013</CopyrightYear></ArticleCopyright><ArticleGrants Type="Regular"><MetadataGrant Grant="OpenAccess"></MetadataGrant><AbstractGrant Grant="OpenAccess"></AbstractGrant><BodyPDFGrant Grant="Restricted"></BodyPDFGrant><BodyHTMLGrant Grant="Restricted"></BodyHTMLGrant><BibliographyGrant Grant="Restricted"></BibliographyGrant><ESMGrant Grant="Restricted"></ESMGrant></ArticleGrants></ArticleInfo><ArticleHeader><AuthorGroup><Author AffiliationIDS="Aff1 Aff2" CorrespondingAffiliationID="Aff1"><AuthorName DisplayOrder="Western"><GivenName>Ilona</GivenName><FamilyName>Stefańska</FamilyName></AuthorName><Contact><Phone>+48-697-497111</Phone><Email>i.stefanska@gmail.com</Email></Contact></Author><Author AffiliationIDS="Aff3"><AuthorName DisplayOrder="Western"><GivenName>Tomasz</GivenName><FamilyName>Dzieciatkowski</FamilyName></AuthorName></Author><Author AffiliationIDS="Aff2 Aff4"><AuthorName DisplayOrder="Western"><GivenName>Lidia</GivenName><GivenName>B.</GivenName><FamilyName>Brydak</FamilyName></AuthorName></Author><Author AffiliationIDS="Aff2"><AuthorName DisplayOrder="Western"><GivenName>Magdalena</GivenName><FamilyName>Romanowska</FamilyName></AuthorName></Author><Affiliation ID="Aff1"><OrgDivision>Department of Fermentation Technology</OrgDivision><OrgName>Institute of Agricultural and Food Biotechnology</OrgName><OrgAddress><Street>36 Rakowiecka Street</Street><Postcode>02-532</Postcode><City>Warsaw</City><Country Code="PL">Poland</Country></OrgAddress></Affiliation><Affiliation ID="Aff2"><OrgDivision>Department of Influenza Research, National Influenza Centre</OrgDivision><OrgName>National Institute of Public Health-National Institute of Hygiene</OrgName><OrgAddress><Street>24 Chocimska Street</Street><Postcode>00-791</Postcode><City>Warsaw</City><Country Code="PL">Poland</Country></OrgAddress></Affiliation><Affiliation ID="Aff3"><OrgDivision>Chair and Department of Medical Microbiology</OrgDivision><OrgName>Medical University of Warsaw</OrgName><OrgAddress><Street>5 Chalubinski Street</Street><Postcode>02-004</Postcode><City>Warsaw</City><Country Code="PL">Poland</Country></OrgAddress></Affiliation><Affiliation ID="Aff4"><OrgDivision>Department of Immunology, Faculty of Biology</OrgDivision><OrgName>University of Szczecin</OrgName><OrgAddress><Street>3a Felczaka Street</Street><Postcode>71-412</Postcode><City>Szczecin</City><Country Code="PL">Poland</Country></OrgAddress></Affiliation></AuthorGroup><Abstract ID="Abs1" Language="En" OutputMedium="All"><Heading>Abstract</Heading><Para>This study was performed to develop real-time PCR (qPCR) for detection of human seasonal and avian influenza viruses in duplex format. First duplex qPCR detects haemagglutinin (HA) gene of influenza virus A(H1N1)pdm09 and HA gene of influenza virus A(H3N2), the second reaction detects neuraminidase (NA) gene of influenza virus A(H3N2) and NA gene of influenza virus A(H1N1)pdm09 and A(H5N1), and the third reaction detects HA gene of influenza A(H5N1) and nonstructural protein gene of influenza B virus. Primers and probes were designed using multiple alignments of target gene sequences of different reference strains. Assays were optimised for identical thermocycling conditions. Their specificity was confirmed by conventional PCR and monoplex qPCR with nucleic acids isolated from different influenza viruses and other respiratory pathogens. Plasmid constructs with a fragment of specific gene were used to assess sensitivity of the assay. The limit of detection ranged from 27 to 96 cDNA copies/reaction. Clinical specimens (n = 107) have been tested using new assays, immunofluorescence and monoplex qRT-PCR. It has been shown that developed assays have been capable of rapid and accurate simultaneous detection and differentiation of influenza viruses. They are more sensitive than immunofluorescence and at least as sensitive as monoplex qRT-PCR.</Para></Abstract></ArticleHeader><NoBody></NoBody></Article></Issue></Volume></Journal></Publisher></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses</title></titleInfo><name type="personal" displayLabel="corresp"><namePart type="given">Ilona</namePart><namePart type="family">Stefańska</namePart><affiliation>Department of Fermentation Technology, Institute of Agricultural and Food Biotechnology, 36 Rakowiecka Street, 02-532, Warsaw, Poland</affiliation><affiliation>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw, Poland</affiliation><affiliation>E-mail: i.stefanska@gmail.com</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Tomasz</namePart><namePart type="family">Dzieciatkowski</namePart><affiliation>Chair and Department of Medical Microbiology, Medical University of Warsaw, 5 Chalubinski Street, 02-004, Warsaw, Poland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Lidia</namePart><namePart type="given">B.</namePart><namePart type="family">Brydak</namePart><affiliation>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw, Poland</affiliation><affiliation>Department of Immunology, Faculty of Biology, University of Szczecin, 3a Felczaka Street, 71-412, Szczecin, Poland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Magdalena</namePart><namePart type="family">Romanowska</namePart><affiliation>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw, Poland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="article" displayLabel="OriginalPaper" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-6N5SZHKN-D">article</genre><originInfo><publisher>Springer Vienna</publisher><place><placeTerm type="text">Vienna</placeTerm></place><dateCreated encoding="w3cdtf">2012-10-22</dateCreated><dateIssued encoding="w3cdtf">2013-08-01</dateIssued><copyrightDate encoding="w3cdtf">2013</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><abstract lang="en">Abstract: This study was performed to develop real-time PCR (qPCR) for detection of human seasonal and avian influenza viruses in duplex format. First duplex qPCR detects haemagglutinin (HA) gene of influenza virus A(H1N1)pdm09 and HA gene of influenza virus A(H3N2), the second reaction detects neuraminidase (NA) gene of influenza virus A(H3N2) and NA gene of influenza virus A(H1N1)pdm09 and A(H5N1), and the third reaction detects HA gene of influenza A(H5N1) and nonstructural protein gene of influenza B virus. Primers and probes were designed using multiple alignments of target gene sequences of different reference strains. Assays were optimised for identical thermocycling conditions. Their specificity was confirmed by conventional PCR and monoplex qPCR with nucleic acids isolated from different influenza viruses and other respiratory pathogens. Plasmid constructs with a fragment of specific gene were used to assess sensitivity of the assay. The limit of detection ranged from 27 to 96 cDNA copies/reaction. Clinical specimens (n = 107) have been tested using new assays, immunofluorescence and monoplex qRT-PCR. It has been shown that developed assays have been capable of rapid and accurate simultaneous detection and differentiation of influenza viruses. They are more sensitive than immunofluorescence and at least as sensitive as monoplex qRT-PCR.</abstract><note>Original Article</note><relatedItem type="host"><titleInfo><title>Archives of Virology</title><subTitle>Official Journal of the Virology Division of the International Union of Microbiological Societies</subTitle></titleInfo><titleInfo type="abbreviated"><title>Arch Virol</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>Springer</publisher><dateIssued encoding="w3cdtf">2013</dateIssued><copyrightDate encoding="w3cdtf">2013</copyrightDate></originInfo><subject><genre>Journal-Subject-Collection</genre><topic authority="SpringerSubjectCodes" authorityURI="SC3">Biomedical and Life Sciences</topic></subject><classification authority="Journal-SubjectCollection-Code">SC3</classification><subject><genre>Journal-Subject-Group</genre><topic authority="SpringerSubjectCodes" authorityURI="SCB">Biomedicine</topic><topic authority="SpringerSubjectCodes" authorityURI="SCB22003">Virology</topic><topic authority="SpringerSubjectCodes" authorityURI="SCB16003">Medical Microbiology</topic><topic authority="SpringerSubjectCodes" authorityURI="SCH33096">Infectious Diseases</topic></subject><identifier type="ISSN">0304-8608</identifier><identifier type="eISSN">1432-8798</identifier><identifier type="JournalID">705</identifier><identifier type="IssueArticleCount">23</identifier><identifier type="VolumeIssueCount">12</identifier><part><date>2013</date><detail type="volume"><number>158</number><caption>vol.</caption></detail><detail type="issue"><number>8</number><caption>no.</caption></detail><extent unit="pages"><start>1743</start><end>1753</end></extent></part><recordInfo><recordOrigin>Springer-Verlag Wien, 2013</recordOrigin></recordInfo></relatedItem><identifier type="istex">7AD255A4E9B6FED2632F9BE8FB3CBFC11CD66951</identifier><identifier type="ark">ark:/67375/VQC-9JNV2659-Q</identifier><identifier type="DOI">10.1007/s00705-013-1648-0</identifier><identifier type="ArticleID">1648</identifier><identifier type="ArticleID">s00705-013-1648-0</identifier><accessCondition type="use and reproduction" contentType="copyright">Springer-Verlag Wien, 2013</accessCondition><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-3XSW68JL-F">springer</recordContentSource><recordOrigin>Converted from (version 1.2.14) to MODS version 3.6.</recordOrigin><recordCreationDate encoding="w3cdtf">2020-05-28</recordCreationDate></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/VQC-9JNV2659-Q/record.json</uri></json:item></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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