Article: Detection of human novel influenza A (H1N1) viruses using multi-fluorescent real-time RT-PCR

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TitleDetection of human novel influenza A (H1N1) viruses using multi-fluorescent real-time RT-PCR
AuthorsDong, H2
Zhang, Y1
Xiong, H2
Yan, A2
Ding, G5
Chen, Y2 4
Xie, L2
Chen, J2
Zhang, G5
Hao, P5
Cong, L1
Lu, Y1
Che, X6
Wang, X7
Li, Y5
Yuen, KY3
Zhao, G2 4 8
Jin, W2 4
Issue Date2010
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/virusres
CitationVirus Research, 2010, v. 147 n. 1, p. 85-90 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.virusres.2009.10.011
AbstractThe novel influenza A (H1N1) virus is now rapidly spreading across the world. Early detection is one of the most effective measures to prevent further transmission of the virus. 4 sets of proprietary primers and probes designed for detection of influenza A viruses (InfA), human and swine H1N1 viruses (SH1), the novel H1N1 viruses (NH1) and RNaseP gene (RP) respectively were pooled together in a single tube for a multi-fluorescent real-time RT-PCR assay. The detection limit was found to be one order more sensitive than that employing the WHO recommended protocol. The NH1 probe was negative for all control samples including human seasonal H1N1 virus, other subtypes of human influenza A viruses (H3, H5, H9), human influenza B virus and nasopharyngeal swabs from patients with noninfluenza respiratory diseases, indicating its high specificity, capable of discriminating the novel influenza A virus from the previously identified H1N1 viruses. For confirmation, the PCR amplified fragment of the hemagglutinin gene was sequenced which could provide enough information to identify the novel H1N1 virus as a distinct cluster among all viruses of subtype H1 through average distance clustering analysis. Although these assays should be useful in the current outbreak for rapid detection and discrimination of the novel H1N1 from swine H1N1 and other human seasonal H1N1 viruses, further design improvement is suggested to match possible sequence variations in the detected region along with the course of the epidemic. © 2009 Elsevier B.V. All rights reserved.
ISSN0168-1702
2011 Impact Factor: 2.941
2011 SCImago Journal Rankings: 0.237
DOIhttp://dx.doi.org/10.1016/j.virusres.2009.10.011
ISI Accession Number IDWOS:000276560300012
ReferencesReferences in Scopus
DC Field
Value
dc.contributor.authorDong, H
dc.contributor.authorZhang, Y
dc.contributor.authorXiong, H
dc.contributor.authorYan, A
dc.contributor.authorDing, G
dc.contributor.authorChen, Y
dc.contributor.authorXie, L
dc.contributor.authorChen, J
dc.contributor.authorZhang, G
dc.contributor.authorHao, P
dc.contributor.authorCong, L
dc.contributor.authorLu, Y
dc.contributor.authorChe, X
dc.contributor.authorWang, X
dc.contributor.authorLi, Y
dc.contributor.authorYuen, KY
dc.contributor.authorZhao, G
dc.contributor.authorJin, W
dc.date.accessioned2012-08-08T08:51:20Z
dc.date.available2012-08-08T08:51:20Z
dc.date.issued2010
dc.description.abstractThe novel influenza A (H1N1) virus is now rapidly spreading across the world. Early detection is one of the most effective measures to prevent further transmission of the virus. 4 sets of proprietary primers and probes designed for detection of influenza A viruses (InfA), human and swine H1N1 viruses (SH1), the novel H1N1 viruses (NH1) and RNaseP gene (RP) respectively were pooled together in a single tube for a multi-fluorescent real-time RT-PCR assay. The detection limit was found to be one order more sensitive than that employing the WHO recommended protocol. The NH1 probe was negative for all control samples including human seasonal H1N1 virus, other subtypes of human influenza A viruses (H3, H5, H9), human influenza B virus and nasopharyngeal swabs from patients with noninfluenza respiratory diseases, indicating its high specificity, capable of discriminating the novel influenza A virus from the previously identified H1N1 viruses. For confirmation, the PCR amplified fragment of the hemagglutinin gene was sequenced which could provide enough information to identify the novel H1N1 virus as a distinct cluster among all viruses of subtype H1 through average distance clustering analysis. Although these assays should be useful in the current outbreak for rapid detection and discrimination of the novel H1N1 from swine H1N1 and other human seasonal H1N1 viruses, further design improvement is suggested to match possible sequence variations in the detected region along with the course of the epidemic. © 2009 Elsevier B.V. All rights reserved.
dc.description.natureLink_to_subscribed_fulltext
dc.identifier.citationVirus Research, 2010, v. 147 n. 1, p. 85-90 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.virusres.2009.10.011
dc.identifier.citeulike6199132
dc.identifier.doihttp://dx.doi.org/10.1016/j.virusres.2009.10.011
dc.identifier.epage90
dc.identifier.isiWOS:000276560300012
dc.identifier.issn0168-1702
2011 Impact Factor: 2.941
2011 SCImago Journal Rankings: 0.237
dc.identifier.issue1
dc.identifier.pmid19883704
dc.identifier.scopuseid_2-s2.0-71349088322
dc.identifier.spage85
dc.identifier.urihttp://hdl.handle.net/10722/157567
dc.identifier.volume147
dc.languageeng
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/virusres
dc.publisher.placeNetherlands
dc.relation.ispartofVirus Research
dc.relation.referencesReferences in Scopus
dc.subject.meshCluster Analysis
dc.subject.meshDna Primers - Genetics
dc.subject.meshGenotype
dc.subject.meshHemagglutinins, Viral - Genetics
dc.subject.meshHumans
dc.subject.meshInfluenza A Virus, H1n1 Subtype - Classification - Genetics - Isolation & Purification
dc.subject.meshInfluenza, Human - Diagnosis - Virology
dc.subject.meshRna, Viral - Genetics
dc.subject.meshReverse Transcriptase Polymerase Chain Reaction - Methods
dc.subject.meshSensitivity And Specificity
dc.subject.meshSequence Analysis, Dna
dc.subject.meshSequence Homology
dc.titleDetection of human novel influenza A (H1N1) viruses using multi-fluorescent real-time RT-PCR
dc.typeArticle
Author Affiliations
  1. Zhejiang Provincial Center for Disease Control and Prevention
  2. Chinese National Human Genome Center at Shanghai
  3. The University of Hong Kong
  4. National Engineering Center for Biochip at Shanghai
  5. Shanghai Institute for Biological Sciences Chinese Academy of Sciences
  6. Zhujiang Hospital
  7. South China University of Technology
  8. Chinese University of Hong Kong