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Article: Design of multiplexed detection assays for identification of avian influenza a virus subtypes pathogenic to humans by SmartCycler real-time reverse transcription-PCR

TitleDesign of multiplexed detection assays for identification of avian influenza a virus subtypes pathogenic to humans by SmartCycler real-time reverse transcription-PCR
Authors
Issue Date2009
PublisherAmerican Society for Microbiology
Citation
Journal Of Clinical Microbiology, 2009, v. 47 n. 1, p. 86-92 How to Cite?
AbstractInfluenza A virus (IAV) epidemics are the result of human-to-human or poultry-to-human transmission. Tracking seasonal outbreaks of IAV and other avian influenza virus (AIV) subtypes that can infect humans, aquatic and migratory birds, poultry, and pigs is essential for epidemiological surveillance and outbreak alerts. In this study, we performed four real-time reverse transcription-PCR (rRT-PCR) assays for identification of the IAV M and hemagglutinin (HA) genes from six known AIVs infecting pigs, birds, and humans. IAV M1 gene-positive samples tested by single-step rRT-PCR and a fluorogenic Sybr green I detection system were further processed for H5 subtype identification by using two-primer-set multiplex and Sybr green I rRT-PCR assays. H5 subtype-negative samples were then tested with either a TaqMan assay for subtypes H1 and H3 or a TaqMan assay for subtypes H2, H7, and H9 and a beacon multiplex rRT-PCR identification assay. The four-tube strategy was able to detect 10 RNA copies of the HA genes of subtypes H1, H2, H3, H5, and H7 and 100 RNA copies of the HA gene of subtype H9. At least six H5 clades of H5N1 viruses isolated in Southeast Asia and China were detected by that test. Using rRT-PCR assays for the M1 and HA genes in 202 nasopharyngeal swab specimens from children with acute respiratory infections, we identified a total of 39 samples positive for the IAV M1 gene and subtypes H1 and H3. When performed with a portable SmartCycler instrument, the assays offer an efficient, flexible, and reliable platform for investigations of IAV and AIV in remote hospitals and in the field. Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Persistent Identifierhttp://hdl.handle.net/10722/59426
ISSN
2015 Impact Factor: 3.631
2015 SCImago Journal Rankings: 2.151
ISI Accession Number ID
Funding AgencyGrant Number
French Ministry of Health
Li Kha Shing Foundation
French Agency for Development (SISEA project)
Hong Kong SARAoE/M-12/06
Funding Information:

The study was supported by the French Ministry of Health, the Li Kha Shing Foundation, the French Agency for Development (SISEA project), and the European Union (RIVERS project). W. W. is a recipient of AREVA from AREVA-Pasteur partnership. J.S.M.P. is supported by the university grants committee of the Hong Kong SAR (project AoE/M-12/06).

References
Grants

 

DC FieldValueLanguage
dc.contributor.authorWang, Wen_HK
dc.contributor.authorRen, Pen_HK
dc.contributor.authorMardi, Sen_HK
dc.contributor.authorHou, Len_HK
dc.contributor.authorTsai, Cen_HK
dc.contributor.authorChan, KHen_HK
dc.contributor.authorCheng, Pen_HK
dc.contributor.authorSheng, Jen_HK
dc.contributor.authorBuchy, Pen_HK
dc.contributor.authorSun, Ben_HK
dc.contributor.authorToyoda, Ten_HK
dc.contributor.authorLim, Wen_HK
dc.contributor.authorPeiris, JSMen_HK
dc.contributor.authorZhou, Pen_HK
dc.contributor.authorDeubel, Ven_HK
dc.date.accessioned2010-05-31T03:49:51Z-
dc.date.available2010-05-31T03:49:51Z-
dc.date.issued2009en_HK
dc.identifier.citationJournal Of Clinical Microbiology, 2009, v. 47 n. 1, p. 86-92en_HK
dc.identifier.issn0095-1137en_HK
dc.identifier.urihttp://hdl.handle.net/10722/59426-
dc.description.abstractInfluenza A virus (IAV) epidemics are the result of human-to-human or poultry-to-human transmission. Tracking seasonal outbreaks of IAV and other avian influenza virus (AIV) subtypes that can infect humans, aquatic and migratory birds, poultry, and pigs is essential for epidemiological surveillance and outbreak alerts. In this study, we performed four real-time reverse transcription-PCR (rRT-PCR) assays for identification of the IAV M and hemagglutinin (HA) genes from six known AIVs infecting pigs, birds, and humans. IAV M1 gene-positive samples tested by single-step rRT-PCR and a fluorogenic Sybr green I detection system were further processed for H5 subtype identification by using two-primer-set multiplex and Sybr green I rRT-PCR assays. H5 subtype-negative samples were then tested with either a TaqMan assay for subtypes H1 and H3 or a TaqMan assay for subtypes H2, H7, and H9 and a beacon multiplex rRT-PCR identification assay. The four-tube strategy was able to detect 10 RNA copies of the HA genes of subtypes H1, H2, H3, H5, and H7 and 100 RNA copies of the HA gene of subtype H9. At least six H5 clades of H5N1 viruses isolated in Southeast Asia and China were detected by that test. Using rRT-PCR assays for the M1 and HA genes in 202 nasopharyngeal swab specimens from children with acute respiratory infections, we identified a total of 39 samples positive for the IAV M1 gene and subtypes H1 and H3. When performed with a portable SmartCycler instrument, the assays offer an efficient, flexible, and reliable platform for investigations of IAV and AIV in remote hospitals and in the field. Copyright © 2009, American Society for Microbiology. All Rights Reserved.en_HK
dc.languageengen_HK
dc.publisherAmerican Society for Microbiologyen_HK
dc.relation.ispartofJournal of Clinical Microbiologyen_HK
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.en_HK
dc.titleDesign of multiplexed detection assays for identification of avian influenza a virus subtypes pathogenic to humans by SmartCycler real-time reverse transcription-PCRen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0095-1137&volume=47&issue=1&spage=86&epage=92&date=2008&atitle=Design+of+multiplexed+detection+assays+for+identification+of+avian+influenza+a+virus+subtypes+pathogenic+to+humans+by+SmartCycler+real-time+reverse+transcription-PCRen_HK
dc.identifier.emailPeiris, JSM: malik@hkucc.hku.hken_HK
dc.identifier.authorityPeiris, JSM=rp00410en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1128/JCM.01090-08en_HK
dc.identifier.pmid18971359-
dc.identifier.scopuseid_2-s2.0-58849091463en_HK
dc.identifier.hkuros166121en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-58849091463&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume47en_HK
dc.identifier.issue1en_HK
dc.identifier.spage86en_HK
dc.identifier.epage92en_HK
dc.identifier.isiWOS:000262158000010-
dc.publisher.placeUnited Statesen_HK
dc.relation.projectControl of Pandemic and Inter-pandemic Influenza-
dc.identifier.scopusauthoridWang, W=36071339100en_HK
dc.identifier.scopusauthoridRen, P=35278694300en_HK
dc.identifier.scopusauthoridMardi, S=36123480900en_HK
dc.identifier.scopusauthoridHou, L=35725433700en_HK
dc.identifier.scopusauthoridTsai, C=26024000800en_HK
dc.identifier.scopusauthoridChan, KH=7406034307en_HK
dc.identifier.scopusauthoridCheng, P=8865849700en_HK
dc.identifier.scopusauthoridSheng, J=35278902400en_HK
dc.identifier.scopusauthoridBuchy, P=15019240000en_HK
dc.identifier.scopusauthoridSun, B=24734369900en_HK
dc.identifier.scopusauthoridToyoda, T=7201723996en_HK
dc.identifier.scopusauthoridLim, W=7202378277en_HK
dc.identifier.scopusauthoridPeiris, JSM=7005486823en_HK
dc.identifier.scopusauthoridZhou, P=7401848639en_HK
dc.identifier.scopusauthoridDeubel, V=7005679225en_HK

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