Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses
Identifieur interne : 001682 ( Main/Merge ); précédent : 001681; suivant : 001683Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses
Auteurs : Ilona Stefa Ska [Pologne] ; Tomasz Dzieciatkowski [Pologne] ; Lidia B. Brydak [Pologne] ; Magdalena Romanowska [Pologne]Source :
- 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.
Url:
DOI: 10.1007/s00705-013-1648-0
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 001807
- to stream Istex, to step Curation: 001807
- to stream Istex, to step Checkpoint: 000524
Links to Exploration step
ISTEX:7AD255A4E9B6FED2632F9BE8FB3CBFC11CD66951Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses</title><author><name sortKey="Stefa Ska, Ilona" sort="Stefa Ska, Ilona" uniqKey="Stefa Ska I" first="Ilona" last="Stefa Ska">Ilona Stefa Ska</name></author><author><name sortKey="Dzieciatkowski, Tomasz" sort="Dzieciatkowski, Tomasz" uniqKey="Dzieciatkowski T" first="Tomasz" last="Dzieciatkowski">Tomasz Dzieciatkowski</name></author><author><name sortKey="Brydak, Lidia B" sort="Brydak, Lidia B" uniqKey="Brydak L" first="Lidia B." last="Brydak">Lidia B. Brydak</name></author><author><name sortKey="Romanowska, Magdalena" sort="Romanowska, Magdalena" uniqKey="Romanowska M" first="Magdalena" last="Romanowska">Magdalena Romanowska</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:7AD255A4E9B6FED2632F9BE8FB3CBFC11CD66951</idno><date when="2013" year="2013">2013</date><idno type="doi">10.1007/s00705-013-1648-0</idno><idno type="url">https://api.istex.fr/ark:/67375/VQC-9JNV2659-Q/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">001807</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001807</idno><idno type="wicri:Area/Istex/Curation">001807</idno><idno type="wicri:Area/Istex/Checkpoint">000524</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">000524</idno><idno type="wicri:doubleKey">0304-8608:2013:Stefa Ska I:application:of:three</idno><idno type="wicri:Area/Main/Merge">001682</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses</title><author><name sortKey="Stefa Ska, Ilona" sort="Stefa Ska, Ilona" uniqKey="Stefa Ska I" first="Ilona" last="Stefa Ska">Ilona Stefa Ska</name><affiliation wicri:level="1"><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Fermentation Technology, Institute of Agricultural and Food Biotechnology, 36 Rakowiecka Street, 02-532, Warsaw</wicri:regionArea><wicri:noRegion>Warsaw</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw</wicri:regionArea><wicri:noRegion>Warsaw</wicri:noRegion></affiliation><affiliation></affiliation></author><author><name sortKey="Dzieciatkowski, Tomasz" sort="Dzieciatkowski, Tomasz" uniqKey="Dzieciatkowski T" first="Tomasz" last="Dzieciatkowski">Tomasz Dzieciatkowski</name><affiliation wicri:level="1"><country xml:lang="fr">Pologne</country><wicri:regionArea>Chair and Department of Medical Microbiology, Medical University of Warsaw, 5 Chalubinski Street, 02-004, Warsaw</wicri:regionArea><wicri:noRegion>Warsaw</wicri:noRegion></affiliation></author><author><name sortKey="Brydak, Lidia B" sort="Brydak, Lidia B" uniqKey="Brydak L" first="Lidia B." last="Brydak">Lidia B. Brydak</name><affiliation wicri:level="1"><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw</wicri:regionArea><wicri:noRegion>Warsaw</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Immunology, Faculty of Biology, University of Szczecin, 3a Felczaka Street, 71-412, Szczecin</wicri:regionArea><wicri:noRegion>Szczecin</wicri:noRegion></affiliation></author><author><name sortKey="Romanowska, Magdalena" sort="Romanowska, Magdalena" uniqKey="Romanowska M" first="Magdalena" last="Romanowska">Magdalena Romanowska</name><affiliation wicri:level="1"><country xml:lang="fr">Pologne</country><wicri:regionArea>Department of Influenza Research, National Influenza Centre, National Institute of Public Health-National Institute of Hygiene, 24 Chocimska Street, 00-791, Warsaw</wicri:regionArea><wicri:noRegion>Warsaw</wicri:noRegion></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></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/PandemieGrippaleV1/Data/Main/Merge
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001682 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Merge/biblio.hfd -nk 001682 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= PandemieGrippaleV1
|flux= Main
|étape= Merge
|type= RBID
|clé= ISTEX:7AD255A4E9B6FED2632F9BE8FB3CBFC11CD66951
|texte= Application of three duplex real-time PCR assays for simultaneous detection of human seasonal and avian influenza viruses
}}
| This area was generated with Dilib version V0.6.34. |  |