Study for establishing differential diagnostic protocols for Japanese encephalitis virus and West Nile virus infection

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In this research program, we established the differential diagnostic protocols for flaviviruses, especially Japanese encephalitis virus (JEV) and West Nile virus (WNV). Because, JEV, which belongs to the same flavivirus serological group, in which WNV belongs, circulates in the nature of Japan, and the symptoms caused by JEV and WNV infections are quite similar, we should differentiate both infections to confirm the impost of WNV into Japan where is a virgin country of WNV infection. (1) We established the flavivirus reverse genetic systems, to produce subviral particles (SvPs), viral replicons and virus-like particles (VLPs). SvPs are the particle structures composed by only viral envelope proteins without viral related RNAs. The viral replicons are the RNAs, which derived from viral defective genomic RNAs and auto-replicate in the replicon-transfected cells. The VLPs are the particles composed of viral envelope proteins as its viral shells, and viral replicons within the shells. (2) We established differential diagnostic protocols, ELISA, IFA and Western blot analysis to differentiate JEV and WNV using SvPs of JEV and WNV. (3) We generated reporter protein-expressing WNV replies. VLPs composed. of JEV or WNV structural proteins as surface envelope proteins, and WNV replicons within the shells, were also generated. We are now establishing a modified-neutralizing test for differential diagnosis of flaviviruses using the VLPs.