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

TitleDetection of human novel influenza A (H1N1) viruses using multi-fluorescent real-time RT-PCR
Authors
Issue Date2010
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/virusres
Citation
Virus Research, 2010, v. 147 n. 1, p. 85-90 How to Cite?
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.
Persistent Identifierhttp://hdl.handle.net/10722/157567
ISSN
2015 Impact Factor: 2.526
2015 SCImago Journal Rankings: 1.259
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorDong, Hen_US
dc.contributor.authorZhang, Yen_US
dc.contributor.authorXiong, Hen_US
dc.contributor.authorYan, Aen_US
dc.contributor.authorDing, Gen_US
dc.contributor.authorChen, Yen_US
dc.contributor.authorXie, Len_US
dc.contributor.authorChen, Jen_US
dc.contributor.authorZhang, Gen_US
dc.contributor.authorHao, Pen_US
dc.contributor.authorCong, Len_US
dc.contributor.authorLu, Yen_US
dc.contributor.authorChe, Xen_US
dc.contributor.authorWang, Xen_US
dc.contributor.authorLi, Yen_US
dc.contributor.authorYuen, KYen_US
dc.contributor.authorZhao, Gen_US
dc.contributor.authorJin, Wen_US
dc.date.accessioned2012-08-08T08:51:20Z-
dc.date.available2012-08-08T08:51:20Z-
dc.date.issued2010en_US
dc.identifier.citationVirus Research, 2010, v. 147 n. 1, p. 85-90en_US
dc.identifier.issn0168-1702en_US
dc.identifier.urihttp://hdl.handle.net/10722/157567-
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.en_US
dc.languageengen_US
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/virusresen_US
dc.relation.ispartofVirus Researchen_US
dc.subject.meshCluster Analysisen_US
dc.subject.meshDna Primers - Geneticsen_US
dc.subject.meshGenotypeen_US
dc.subject.meshHemagglutinins, Viral - Geneticsen_US
dc.subject.meshHumansen_US
dc.subject.meshInfluenza A Virus, H1n1 Subtype - Classification - Genetics - Isolation & Purificationen_US
dc.subject.meshInfluenza, Human - Diagnosis - Virologyen_US
dc.subject.meshRna, Viral - Geneticsen_US
dc.subject.meshReverse Transcriptase Polymerase Chain Reaction - Methodsen_US
dc.subject.meshSensitivity And Specificityen_US
dc.subject.meshSequence Analysis, Dnaen_US
dc.subject.meshSequence Homologyen_US
dc.titleDetection of human novel influenza A (H1N1) viruses using multi-fluorescent real-time RT-PCRen_US
dc.typeArticleen_US
dc.identifier.emailYuen, KY:kyyuen@hkucc.hku.hken_US
dc.identifier.authorityYuen, KY=rp00366en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.virusres.2009.10.011en_US
dc.identifier.pmid19883704-
dc.identifier.scopuseid_2-s2.0-71349088322en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-71349088322&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume147en_US
dc.identifier.issue1en_US
dc.identifier.spage85en_US
dc.identifier.epage90en_US
dc.identifier.isiWOS:000276560300012-
dc.publisher.placeNetherlandsen_US
dc.identifier.scopusauthoridDong, H=35770452600en_US
dc.identifier.scopusauthoridZhang, Y=7601335835en_US
dc.identifier.scopusauthoridXiong, H=36852326000en_US
dc.identifier.scopusauthoridYan, A=35172377100en_US
dc.identifier.scopusauthoridDing, G=35193643700en_US
dc.identifier.scopusauthoridChen, Y=35232572900en_US
dc.identifier.scopusauthoridXie, L=35489434700en_US
dc.identifier.scopusauthoridChen, J=35486923100en_US
dc.identifier.scopusauthoridZhang, G=14008073500en_US
dc.identifier.scopusauthoridHao, P=7005522275en_US
dc.identifier.scopusauthoridCong, L=36896912700en_US
dc.identifier.scopusauthoridLu, Y=26427705800en_US
dc.identifier.scopusauthoridChe, X=7005743182en_US
dc.identifier.scopusauthoridWang, X=11939400000en_US
dc.identifier.scopusauthoridLi, Y=9274991700en_US
dc.identifier.scopusauthoridYuen, KY=36078079100en_US
dc.identifier.scopusauthoridZhao, G=7403296532en_US
dc.identifier.scopusauthoridJin, W=7402071191en_US
dc.identifier.citeulike6199132-

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