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

日本脑炎病毒与西尼罗病毒感染鉴别诊断方案的建立研究

基本信息

批准号：
18580301
负责人：
MAEDA Akihiko
金额：
$ 2.38万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

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.

在该研究计划中，我们建立了黄酮病毒的鉴别诊断方案，尤其是日本脑炎病毒（JEV）和西尼罗河病毒（WNV）。因为，属于WNV属于同一黄病毒血清学群体的JEV在日本的性质中循环，而JEV和WNV感染引起的症状非常相似，我们应该区分这两种感染以确认WNV进入日本的Impost in wnv wnv感染的处女。（1）我们建立了黄病毒反向遗传系统，以产生皮下颗粒（SVP），病毒复制子和病毒样颗粒（VLP）。 SVP是仅由没有病毒相关RNA的病毒包膜蛋白组成的粒子结构。病毒复制子是RNA，它源自复制子转染的细胞中的病毒缺陷基因组RNA和自身重复。 VLP是由病毒包膜蛋白作为病毒壳组成的颗粒，在壳内部的病毒复制子组成。（2）我们使用JEV和WNV的SVP建立了差异诊断方案，ELISA，IFA和Western印迹分析，以区分JEV和WNV。（3）我们生成了表达报告蛋白的WNV答复。 VLP组成。还产生了JEV或WNV结构蛋白作为表面包膜蛋白和壳内的WNV复制子的蛋白质的。现在，我们正在建立一个修改的中和测试，用于使用VLP对黄病毒的鉴别诊断。