A multiplex one-step real-time RT-PCR assay for influenza surveillance.
Identifieur interne : 000309 ( Main/Exploration ); précédent : 000308; suivant : 000310A multiplex one-step real-time RT-PCR assay for influenza surveillance.
Auteurs : I. Huber [Allemagne] ; H. Campe ; D. Sebah ; C. Hartberger ; R. Konrad ; M. Bayer ; U. Busch ; A. SingSource :
- Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin [ 1560-7917 ] ; 2011.
Descripteurs français
- KwdFr :
- ARN viral (génétique), Allemagne (MeSH), Amorces ADN (génétique), Amplification de gène (MeSH), Grippe humaine (diagnostic), Grippe humaine (virologie), Gènes viraux (MeSH), Humains (MeSH), RT-PCR (méthodes), Sensibilité et spécificité (MeSH), Sous-type H1N1 du virus de la grippe A (génétique), Surveillance de la population (MeSH), Trousses de réactifs pour diagnostic (MeSH), Virus de la grippe A (génétique), Virus de la grippe A (isolement et purification), Virus influenza B (génétique), Virus influenza B (isolement et purification).
- MESH :
- diagnostic : Grippe humaine.
- génétique : ARN viral, Amorces ADN, Sous-type H1N1 du virus de la grippe A, Virus de la grippe A, Virus influenza B.
- isolement et purification : Virus de la grippe A, Virus influenza B.
- méthodes : RT-PCR.
- virologie : Grippe humaine.
- Allemagne, Amplification de gène, Gènes viraux, Humains, Sensibilité et spécificité, Surveillance de la population, Trousses de réactifs pour diagnostic.
- Wicri :
- geographic : Allemagne.
English descriptors
- KwdEn :
- DNA Primers (genetics), Gene Amplification (MeSH), Genes, Viral (MeSH), Germany (MeSH), Humans (MeSH), Influenza A Virus, H1N1 Subtype (genetics), Influenza A virus (genetics), Influenza A virus (isolation & purification), Influenza B virus (genetics), Influenza B virus (isolation & purification), Influenza, Human (diagnosis), Influenza, Human (virology), Population Surveillance (MeSH), RNA, Viral (genetics), Reagent Kits, Diagnostic (MeSH), Reverse Transcriptase Polymerase Chain Reaction (methods), Sensitivity and Specificity (MeSH).
- MESH :
- chemical , genetics : DNA Primers, RNA, Viral.
- geographic : Germany, Reagent Kits, Diagnostic.
- diagnosis : Influenza, Human.
- genetics : Influenza A Virus, H1N1 Subtype, Influenza A virus, Influenza B virus.
- isolation & purification : Influenza A virus, Influenza B virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction.
- virology : Influenza, Human.
- Gene Amplification, Genes, Viral, Humans, Population Surveillance, Sensitivity and Specificity.
Abstract
For surveillance purposes real-time PCR assays for influenza viruses had to be adapted to the pandemic influenza A(H1N1)2009 strain. We combined published primers and probes for influenza A, influenza B and an internal amplification control with a detection system for influenza A(H1N1)2009 to set up a rapid, reliable, simple and cost-effective high-throughput multiplex one-step real-time RT-PCR. The workflow also includes automated sample preparation for high-throughput screening. The lower limit of detection of the multiplex assay was 3.5x10(2) RNA copies per PCR reaction. The diagnostic sensitivity of the multiplex assay was 87.7%, but increased to 99.4% for influenza-positive samples yielding C(t) values of less than 34 cycles in the respective diagnostic assay. High specificity was confirmed by sequencing and correct detection of 15 reference samples from two quality assurance studies. The multiplex PCR was introduced for surveillance of samples from a network of general practitioners and paediatricians in Bavaria, Germany during the influenza pandemic of 2009. Comparison with surveillance data from reported cases proved the reliability of the multiplex assay for influenza surveillance programmes.
PubMed: 21345319
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">A multiplex one-step real-time RT-PCR assay for influenza surveillance.</title><author><name sortKey="Huber, I" sort="Huber, I" uniqKey="Huber I" first="I" last="Huber">I. Huber</name><affiliation wicri:level="1"><nlm:affiliation>Bavarian Health and Food Safety Authority, LGL, Oberschleibheim, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Bavarian Health and Food Safety Authority, LGL, Oberschleibheim</wicri:regionArea><wicri:noRegion>Oberschleibheim</wicri:noRegion><wicri:noRegion>Oberschleibheim</wicri:noRegion><wicri:noRegion>Oberschleibheim</wicri:noRegion></affiliation></author><author><name sortKey="Campe, H" sort="Campe, H" uniqKey="Campe H" first="H" last="Campe">H. Campe</name></author><author><name sortKey="Sebah, D" sort="Sebah, D" uniqKey="Sebah D" first="D" last="Sebah">D. Sebah</name></author><author><name sortKey="Hartberger, C" sort="Hartberger, C" uniqKey="Hartberger C" first="C" last="Hartberger">C. Hartberger</name></author><author><name sortKey="Konrad, R" sort="Konrad, R" uniqKey="Konrad R" first="R" last="Konrad">R. Konrad</name></author><author><name sortKey="Bayer, M" sort="Bayer, M" uniqKey="Bayer M" first="M" last="Bayer">M. Bayer</name></author><author><name sortKey="Busch, U" sort="Busch, U" uniqKey="Busch U" first="U" last="Busch">U. Busch</name></author><author><name sortKey="Sing, A" sort="Sing, A" uniqKey="Sing A" first="A" last="Sing">A. Sing</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2011">2011</date><idno type="RBID">pubmed:21345319</idno><idno type="pmid">21345319</idno><idno type="wicri:Area/Main/Corpus">000297</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000297</idno><idno type="wicri:Area/Main/Curation">000297</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000297</idno><idno type="wicri:Area/Main/Exploration">000297</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">A multiplex one-step real-time RT-PCR assay for influenza surveillance.</title><author><name sortKey="Huber, I" sort="Huber, I" uniqKey="Huber I" first="I" last="Huber">I. Huber</name><affiliation wicri:level="1"><nlm:affiliation>Bavarian Health and Food Safety Authority, LGL, Oberschleibheim, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Bavarian Health and Food Safety Authority, LGL, Oberschleibheim</wicri:regionArea><wicri:noRegion>Oberschleibheim</wicri:noRegion><wicri:noRegion>Oberschleibheim</wicri:noRegion><wicri:noRegion>Oberschleibheim</wicri:noRegion></affiliation></author><author><name sortKey="Campe, H" sort="Campe, H" uniqKey="Campe H" first="H" last="Campe">H. Campe</name></author><author><name sortKey="Sebah, D" sort="Sebah, D" uniqKey="Sebah D" first="D" last="Sebah">D. Sebah</name></author><author><name sortKey="Hartberger, C" sort="Hartberger, C" uniqKey="Hartberger C" first="C" last="Hartberger">C. Hartberger</name></author><author><name sortKey="Konrad, R" sort="Konrad, R" uniqKey="Konrad R" first="R" last="Konrad">R. Konrad</name></author><author><name sortKey="Bayer, M" sort="Bayer, M" uniqKey="Bayer M" first="M" last="Bayer">M. Bayer</name></author><author><name sortKey="Busch, U" sort="Busch, U" uniqKey="Busch U" first="U" last="Busch">U. Busch</name></author><author><name sortKey="Sing, A" sort="Sing, A" uniqKey="Sing A" first="A" last="Sing">A. Sing</name></author></analytic><series><title level="j">Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin</title><idno type="eISSN">1560-7917</idno><imprint><date when="2011" type="published">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>DNA Primers (genetics)</term><term>Gene Amplification (MeSH)</term><term>Genes, Viral (MeSH)</term><term>Germany (MeSH)</term><term>Humans (MeSH)</term><term>Influenza A Virus, H1N1 Subtype (genetics)</term><term>Influenza A virus (genetics)</term><term>Influenza A virus (isolation & purification)</term><term>Influenza B virus (genetics)</term><term>Influenza B virus (isolation & purification)</term><term>Influenza, Human (diagnosis)</term><term>Influenza, Human (virology)</term><term>Population Surveillance (MeSH)</term><term>RNA, Viral (genetics)</term><term>Reagent Kits, Diagnostic (MeSH)</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term><term>Sensitivity and Specificity (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (génétique)</term><term>Allemagne (MeSH)</term><term>Amorces ADN (génétique)</term><term>Amplification de gène (MeSH)</term><term>Grippe humaine (diagnostic)</term><term>Grippe humaine (virologie)</term><term>Gènes viraux (MeSH)</term><term>Humains (MeSH)</term><term>RT-PCR (méthodes)</term><term>Sensibilité et spécificité (MeSH)</term><term>Sous-type H1N1 du virus de la grippe A (génétique)</term><term>Surveillance de la population (MeSH)</term><term>Trousses de réactifs pour diagnostic (MeSH)</term><term>Virus de la grippe A (génétique)</term><term>Virus de la grippe A (isolement et purification)</term><term>Virus influenza B (génétique)</term><term>Virus influenza B (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA Primers</term><term>RNA, Viral</term></keywords><keywords scheme="MESH" type="geographic" xml:lang="en"><term>Germany</term><term>Reagent Kits, Diagnostic</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Influenza, Human</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Grippe humaine</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Influenza A Virus, H1N1 Subtype</term><term>Influenza A virus</term><term>Influenza B virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN viral</term><term>Amorces ADN</term><term>Sous-type H1N1 du virus de la grippe A</term><term>Virus de la grippe A</term><term>Virus influenza B</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Influenza A virus</term><term>Influenza B virus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Virus de la grippe A</term><term>Virus influenza B</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Reverse Transcriptase Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>RT-PCR</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Grippe humaine</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Influenza, Human</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Amplification</term><term>Genes, Viral</term><term>Humans</term><term>Population Surveillance</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Allemagne</term><term>Amplification de gène</term><term>Gènes viraux</term><term>Humains</term><term>Sensibilité et spécificité</term><term>Surveillance de la population</term><term>Trousses de réactifs pour diagnostic</term></keywords><keywords scheme="Wicri" type="geographic" xml:lang="fr"><term>Allemagne</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">For surveillance purposes real-time PCR assays for influenza viruses had to be adapted to the pandemic influenza A(H1N1)2009 strain. We combined published primers and probes for influenza A, influenza B and an internal amplification control with a detection system for influenza A(H1N1)2009 to set up a rapid, reliable, simple and cost-effective high-throughput multiplex one-step real-time RT-PCR. The workflow also includes automated sample preparation for high-throughput screening. The lower limit of detection of the multiplex assay was 3.5x10(2) RNA copies per PCR reaction. The diagnostic sensitivity of the multiplex assay was 87.7%, but increased to 99.4% for influenza-positive samples yielding C(t) values of less than 34 cycles in the respective diagnostic assay. High specificity was confirmed by sequencing and correct detection of 15 reference samples from two quality assurance studies. The multiplex PCR was introduced for surveillance of samples from a network of general practitioners and paediatricians in Bavaria, Germany during the influenza pandemic of 2009. Comparison with surveillance data from reported cases proved the reliability of the multiplex assay for influenza surveillance programmes.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">21345319</PMID><DateCompleted><Year>2011</Year><Month>04</Month><Day>14</Day></DateCompleted><DateRevised><Year>2013</Year><Month>05</Month><Day>01</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1560-7917</ISSN><JournalIssue CitedMedium="Internet"><Volume>16</Volume><Issue>7</Issue><PubDate><Year>2011</Year><Month>Feb</Month><Day>17</Day></PubDate></JournalIssue><Title>Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin</Title><ISOAbbreviation>Euro Surveill.</ISOAbbreviation></Journal><ArticleTitle>A multiplex one-step real-time RT-PCR assay for influenza surveillance.</ArticleTitle><ELocationID EIdType="pii" ValidYN="Y">19798</ELocationID><Abstract><AbstractText>For surveillance purposes real-time PCR assays for influenza viruses had to be adapted to the pandemic influenza A(H1N1)2009 strain. We combined published primers and probes for influenza A, influenza B and an internal amplification control with a detection system for influenza A(H1N1)2009 to set up a rapid, reliable, simple and cost-effective high-throughput multiplex one-step real-time RT-PCR. The workflow also includes automated sample preparation for high-throughput screening. The lower limit of detection of the multiplex assay was 3.5x10(2) RNA copies per PCR reaction. The diagnostic sensitivity of the multiplex assay was 87.7%, but increased to 99.4% for influenza-positive samples yielding C(t) values of less than 34 cycles in the respective diagnostic assay. High specificity was confirmed by sequencing and correct detection of 15 reference samples from two quality assurance studies. The multiplex PCR was introduced for surveillance of samples from a network of general practitioners and paediatricians in Bavaria, Germany during the influenza pandemic of 2009. Comparison with surveillance data from reported cases proved the reliability of the multiplex assay for influenza surveillance programmes.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Huber</LastName><ForeName>I</ForeName><Initials>I</Initials><AffiliationInfo><Affiliation>Bavarian Health and Food Safety Authority, LGL, Oberschleibheim, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Campe</LastName><ForeName>H</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Sebah</LastName><ForeName>D</ForeName><Initials>D</Initials></Author><Author ValidYN="Y"><LastName>Hartberger</LastName><ForeName>C</ForeName><Initials>C</Initials></Author><Author ValidYN="Y"><LastName>Konrad</LastName><ForeName>R</ForeName><Initials>R</Initials></Author><Author ValidYN="Y"><LastName>Bayer</LastName><ForeName>M</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Busch</LastName><ForeName>U</ForeName><Initials>U</Initials></Author><Author ValidYN="Y"><LastName>Sing</LastName><ForeName>A</ForeName><Initials>A</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2011</Year><Month>02</Month><Day>17</Day></ArticleDate></Article><MedlineJournalInfo><Country>Sweden</Country><MedlineTA>Euro Surveill</MedlineTA><NlmUniqueID>100887452</NlmUniqueID><ISSNLinking>1025-496X</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D017931">DNA Primers</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D012367">RNA, Viral</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011933">Reagent Kits, Diagnostic</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D017931" MajorTopicYN="N">DNA Primers</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005784" MajorTopicYN="N">Gene Amplification</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005814" MajorTopicYN="N">Genes, Viral</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005858" MajorTopicYN="N" Type="Geographic">Germany</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D053118" MajorTopicYN="N">Influenza A Virus, H1N1 Subtype</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009980" MajorTopicYN="N">Influenza A virus</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009981" MajorTopicYN="N">Influenza B virus</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D007251" MajorTopicYN="N">Influenza, Human</DescriptorName><QualifierName UI="Q000175" MajorTopicYN="Y">diagnosis</QualifierName><QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011159" MajorTopicYN="Y">Population Surveillance</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012367" MajorTopicYN="N">RNA, Viral</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011933" MajorTopicYN="N">Reagent Kits, Diagnostic</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020133" MajorTopicYN="N">Reverse Transcriptase Polymerase Chain Reaction</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012680" MajorTopicYN="N">Sensitivity and Specificity</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2011</Year><Month>2</Month><Day>25</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2011</Year><Month>2</Month><Day>25</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2011</Year><Month>4</Month><Day>16</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">21345319</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Allemagne</li></country></list><tree><noCountry><name sortKey="Bayer, M" sort="Bayer, M" uniqKey="Bayer M" first="M" last="Bayer">M. Bayer</name><name sortKey="Busch, U" sort="Busch, U" uniqKey="Busch U" first="U" last="Busch">U. Busch</name><name sortKey="Campe, H" sort="Campe, H" uniqKey="Campe H" first="H" last="Campe">H. Campe</name><name sortKey="Hartberger, C" sort="Hartberger, C" uniqKey="Hartberger C" first="C" last="Hartberger">C. Hartberger</name><name sortKey="Konrad, R" sort="Konrad, R" uniqKey="Konrad R" first="R" last="Konrad">R. Konrad</name><name sortKey="Sebah, D" sort="Sebah, D" uniqKey="Sebah D" first="D" last="Sebah">D. Sebah</name><name sortKey="Sing, A" sort="Sing, A" uniqKey="Sing A" first="A" last="Sing">A. Sing</name></noCountry><country name="Allemagne"><noRegion><name sortKey="Huber, I" sort="Huber, I" uniqKey="Huber I" first="I" last="Huber">I. Huber</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/GrippeAllemagneV4/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000309 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 000309 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= GrippeAllemagneV4
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:21345319
|texte= A multiplex one-step real-time RT-PCR assay for influenza surveillance.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:21345319" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a GrippeAllemagneV4
| This area was generated with Dilib version V0.6.35. |  |