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

摘要: Abstract West Nile virus (WNV) and Japanese encephalitis (JEV) are important mosquito-borne viruses of the Flaviviridae family, associated with encephalitis, mainly in humans horses. WNV is also pathogen for many bird species. The incidence human animal infections Europe has risen, mostly recent years, JEV was detected 2011 mosquitoes collected Italy may emerge same way as other flaviviruses had emerged recently (USUTU Bagaza virus) should be regarded a potential threat to public health. Prompt identification discrimination between provides critical epidemiological data prevalence studies health management policies. Here we describe quantitative one-step duplex TaqMan RT-PCR, targeting non-structural protein 2A gene (NS2A-qRT-PCR), based on only one primer pair two probes differential diagnosis JEV. Also this assay enables detection both lineages (WNV-1 WNV-2). To access specificity NS2A-qRT-PCR panel different arboviruses were used. shown specific WNV-2), related Kunjin JEV, since no cross-reactions observed tested arboviruses. Sensitivity determined using serial dilutions vitro-transcribed RNA from very sensitive, being able detect 10 copies RNA. This suitable tool valuable addition methods currently available routine these zoonoses surveillance studies.