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Simultaneous detection of West Nile and Japanese encephalitis virus RNA by duplex TaqMan RT-PCR

2013-11Journal article Restricted access
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dc.contributor.authorBarros, S.C.
dc.contributor.authorRamos, F.
dc.contributor.authorZé-Zé, Líbia
dc.contributor.authorAlves, M.J.
dc.contributor.authorFagulha, T.
dc.contributor.authorDuarte, M.
dc.contributor.authorHenriques, M.
dc.contributor.authorLuís, T.
dc.contributor.authorFevereiro, M.
dc.date.accessioned2013-10-22T12:05:41Z
dc.date.available2013-10-22T12:05:41Z
dc.date.issued2013-11
dc.description.abstractWest Nile virus (WNV) and Japanese encephalitis virus (JEV) are important mosquito-borne viruses of the Flaviviridae family, associated with encephalitis, mainly in humans and horses. WNV is also pathogen for many bird species. The incidence of human and animal WNV infections in Europe has risen, mostly in recent years, and JEV was detected in 2011 in mosquitoes collected in Italy and may emerge in Europe in the same way as other flaviviruses had emerged recently (USUTU and Bagaza virus) and should be regarded as a potential threat to public health. Prompt identification and discrimination between WNV and JEV provides critical epidemiological data for prevalence studies and public and animal health management policies. Here we describe a quantitative one-step duplex TaqMan RT-PCR, targeting non-structural protein 2A gene (NS2A-qRT-PCR), based on only one primer pair and two probes for differential diagnosis of WNV and JEV. Also this assay enables the detection of both WNV lineages (WNV-1 and WNV-2). To access the specificity of NS2A-qRT-PCR a panel of different arboviruses were used. The assay was shown to be specific for both WNV lineages (WNV-1 and WNV-2), WNV related Kunjin virus and JEV, since no cross-reactions were observed with other tested arboviruses. Sensitivity of the assay was determined using serial dilutions of in vitro-transcribed RNA from WNV and JEV. The duplex NS2A-qRT-PCR assay was shown to be very sensitive, being able to detect 10 copies of WNV and JEV RNA. This assay is a suitable tool for the diagnosis of WNV and JEV, and provides a valuable addition to the methods currently available for routine diagnosis of these zoonoses and for surveillance studies.por
dc.identifier.citationJ Virol Methods. 2013 Nov;193(2):554-7. doi: 10.1016/j.jviromet.2013.07.025. Epub 2013 Jul 25.por
dc.identifier.issn0166-0934
dc.identifier.otherdoi: 10.1016/j.jviromet.2013.07.025
dc.identifier.urihttp://hdl.handle.net/10400.18/1767
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherElsevier B.V.por
dc.relation.publisherversionhttp://www.sciencedirect.com/science/article/pii/S0166093413002796por
dc.subjectInfecções Sistémicas e Zoonosespor
dc.subjectWNVpor
dc.subjectJEVpor
dc.subjectDiagnosispor
dc.subjectDuplex RT-PCRpor
dc.titleSimultaneous detection of West Nile and Japanese encephalitis virus RNA by duplex TaqMan RT-PCRpor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage557por
oaire.citation.startPage554por
oaire.citation.titleJournal of Virological Methodspor
oaire.citation.volume193(2)por
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor

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