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Article: Molecular detection of a novel human influenza (H1N1) of pandemic potential by conventional and real-time quantitative RT-PCR assays

TitleMolecular detection of a novel human influenza (H1N1) of pandemic potential by conventional and real-time quantitative RT-PCR assays
Authors
Issue Date2009
PublisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.org
Citation
Clinical Chemistry, 2009, v. 55 n. 8, p. 1555-1558 How to Cite?
AbstractBACKGROUND: Influenza A viruses are medically important viral pathogens that cause significant mortality and morbidity throughout the world. The recent emergence of a novel human influenza A virus (H1N1) poses a serious health threat. Molecular tests for rapid detection of this virus are urgently needed. METHODS: We developed a conventional 1-step RT-PCR assay and a 1-step quantitative real-time RT-PCR assay to detect the novel H1N1 virus, but not the seasonal H1N1 viruses. We also developed an additional real-time RTPCR that can discriminate the novel H1N1 from other swine and human H1 subtype viruses. RESULTS: All of the assays had detection limits for the positive control in the range of 1.0 x 10-4 to 2.0 x 10-3 of the median tissue culture infective dose. Assay specificities were high, and for the conventional and real-time assays, all negative control samples were negative, including 7 human seasonal H1N1 viruses, 1 human H2N2 virus, 2 human seasonal H3N2 viruses, 1 human H5N1 virus, 7 avian influenza viruses (HA subtypes 4, 5, 7, 8, 9, and 10), and 48 nasopharyngeal aspirates (NPAs) from patients with noninfluenza respiratory diseases; for the assay that discriminates the novel H1N1 from other swine and human H1 subtype viruses, all negative controls were also negative, including 20 control NPAs, 2 seasonal human H1N1 viruses, 2 seasonal human H3N2 viruses, and 2 human H5N1 viruses. CONCLUSIONS: These assays appear useful for the rapid diagnosis of cases with the novel H1N1 virus, thereby allowing better pandemic preparedness.
Persistent Identifierhttp://hdl.handle.net/10722/59401
ISSN
2023 Impact Factor: 7.1
2023 SCImago Journal Rankings: 1.460
ISI Accession Number ID
Funding AgencyGrant Number
Area of Excellence Scheme of the University Grants Committee Hong Kong GrantAoE/M-12/06
Research Grant Council of Hong KongHKU 773408M
Funding Information:

Research Funding: Area of Excellence Scheme of the University Grants Committee Hong Kong Grant AoE/M-12/06 and Research Grant Council of Hong Kong (HKU 773408M, LLM).

References
Grants

 

DC FieldValueLanguage
dc.contributor.authorPoon, LLMen_HK
dc.contributor.authorChan, KHen_HK
dc.contributor.authorSmith, GJen_HK
dc.contributor.authorLeung, CSWen_HK
dc.contributor.authorGuan, Yen_HK
dc.contributor.authorYuen, KYen_HK
dc.contributor.authorPeiris, JSMen_HK
dc.date.accessioned2010-05-31T03:49:21Z-
dc.date.available2010-05-31T03:49:21Z-
dc.date.issued2009en_HK
dc.identifier.citationClinical Chemistry, 2009, v. 55 n. 8, p. 1555-1558en_HK
dc.identifier.issn0009-9147en_HK
dc.identifier.urihttp://hdl.handle.net/10722/59401-
dc.description.abstractBACKGROUND: Influenza A viruses are medically important viral pathogens that cause significant mortality and morbidity throughout the world. The recent emergence of a novel human influenza A virus (H1N1) poses a serious health threat. Molecular tests for rapid detection of this virus are urgently needed. METHODS: We developed a conventional 1-step RT-PCR assay and a 1-step quantitative real-time RT-PCR assay to detect the novel H1N1 virus, but not the seasonal H1N1 viruses. We also developed an additional real-time RTPCR that can discriminate the novel H1N1 from other swine and human H1 subtype viruses. RESULTS: All of the assays had detection limits for the positive control in the range of 1.0 x 10-4 to 2.0 x 10-3 of the median tissue culture infective dose. Assay specificities were high, and for the conventional and real-time assays, all negative control samples were negative, including 7 human seasonal H1N1 viruses, 1 human H2N2 virus, 2 human seasonal H3N2 viruses, 1 human H5N1 virus, 7 avian influenza viruses (HA subtypes 4, 5, 7, 8, 9, and 10), and 48 nasopharyngeal aspirates (NPAs) from patients with noninfluenza respiratory diseases; for the assay that discriminates the novel H1N1 from other swine and human H1 subtype viruses, all negative controls were also negative, including 20 control NPAs, 2 seasonal human H1N1 viruses, 2 seasonal human H3N2 viruses, and 2 human H5N1 viruses. CONCLUSIONS: These assays appear useful for the rapid diagnosis of cases with the novel H1N1 virus, thereby allowing better pandemic preparedness.en_HK
dc.languageengen_HK
dc.publisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.orgen_HK
dc.relation.ispartofClinical Chemistryen_HK
dc.subject.meshAnimalsen_HK
dc.subject.meshBase Sequenceen_HK
dc.subject.meshDNA, Viral - analysis - geneticsen_HK
dc.subject.meshHumansen_HK
dc.subject.meshInfluenza A Virus, H1N1 Subtype - classification - genetics - isolation & purificationen_HK
dc.subject.meshInfluenza A Virus, H2N2 Subtype - classification - genetics - isolation & purificationen_HK
dc.subject.meshInfluenza A Virus, H3N2 Subtype - classification - genetics - isolation & purificationen_HK
dc.subject.meshInfluenza A Virus, H5N1 Subtype - classification - genetics - isolation & purificationen_HK
dc.subject.meshInfluenza, Human - diagnosis - virologyen_HK
dc.subject.meshMolecular Diagnostic Techniques - economics - methodsen_HK
dc.subject.meshOrthomyxoviridae Infections - diagnosis - virologyen_HK
dc.subject.meshRNA, Viral - analysis - genetics - isolation & purificationen_HK
dc.subject.meshReverse Transcriptase Polymerase Chain Reaction - economics - methodsen_HK
dc.subject.meshSensitivity and Specificityen_HK
dc.subject.meshSwine - virologyen_HK
dc.subject.meshTime Factorsen_HK
dc.titleMolecular detection of a novel human influenza (H1N1) of pandemic potential by conventional and real-time quantitative RT-PCR assaysen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0009-9147&volume=55&spage=1555&epage=1558&date=2009&atitle=Molecular+Detection+of+a+Novel+Human+Influenza+(H1N1)+of+Pandemic+Potential+by+Conventional+and+Real-Time+Quantitative+RT-PCR+Assaysen_HK
dc.identifier.emailPoon, LLM:llmpoon@hkucc.hku.hken_HK
dc.identifier.emailSmith, GJ:gjsmith@hkucc.hku.hken_HK
dc.identifier.emailGuan, Y:yguan@hkucc.hku.hken_HK
dc.identifier.emailYuen, KY:kyyuen@hkucc.hku.hken_HK
dc.identifier.emailPeiris, JSM:malik@hkucc.hku.hken_HK
dc.identifier.authorityPoon, LLM=rp00484en_HK
dc.identifier.authoritySmith, GJ=rp00444en_HK
dc.identifier.authorityGuan, Y=rp00397en_HK
dc.identifier.authorityYuen, KY=rp00366en_HK
dc.identifier.authorityPeiris, JSM=rp00410en_HK
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1373/clinchem.2009.130229en_HK
dc.identifier.pmid19439731-
dc.identifier.scopuseid_2-s2.0-67649233244en_HK
dc.identifier.hkuros160811en_HK
dc.identifier.hkuros164356-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-67649233244&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume55en_HK
dc.identifier.issue8en_HK
dc.identifier.spage1555en_HK
dc.identifier.epage1558en_HK
dc.identifier.eissn1530-8561-
dc.identifier.isiWOS:000268557500016-
dc.publisher.placeUnited Statesen_HK
dc.relation.projectCodon biases of avian and human influenza A viruses-
dc.relation.projectControl of Pandemic and Inter-pandemic Influenza-
dc.identifier.scopusauthoridPoon, LLM=7005441747en_HK
dc.identifier.scopusauthoridChan, KH=35338760600en_HK
dc.identifier.scopusauthoridSmith, GJ=8344015800en_HK
dc.identifier.scopusauthoridLeung, CSW=7402612654en_HK
dc.identifier.scopusauthoridGuan, Y=7202924055en_HK
dc.identifier.scopusauthoridYuen, KY=36078079100en_HK
dc.identifier.scopusauthoridPeiris, JSM=7005486823en_HK
dc.identifier.issnl0009-9147-

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