一株西尼罗病毒株的生物学特征研究  

The biological characteristics of a West Nile virus strain

作　　者：张久松[1] 张泮河[1] 詹琳[1] 司炳银[1] 祝庆余[1] 曹务春[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100071

出　　处：《中华微生物学和免疫学杂志》2005年第2期133-136,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家博士后科研基金资助项目 (2 0 0 3 0 3 3 196) ;国家科技攻关计划课题 (2 0 0 3BA712A .0 8 0 1) ;国家自然科学基金资助项目(3 0 4714 95 )

摘　　要：目的　对引进的西尼罗病毒 (WNV)毒株的形态学、致病性、细胞敏感性、免疫原性等生物学性状进行探讨 ,为进一步开展WNV的相关研究奠定基础。方法　选择Vero E6与C6 36细胞观察WNV的细胞病变效应 (CPE) ,并制作电镜负染标本和组织切片标本进行病毒形态学鉴定 ;通过乳鼠脑内接种WNV确认其致病性 ;以灭活的感染乳鼠脑悬液免疫BALB c小鼠 ,以间接免疫荧光试验(IFA)测定其血清WNVIgG抗体 ;用RT PCR检测病毒核酸 ,扩增的核苷酸序列通过BLAST作同源性比较。结果　WNV所致Vero E6与C6 36细胞的CPE分别以细胞圆缩和融合为主要特征 ;电镜下所见病毒体为有包膜、直径约 30～ 5 0nm的球形颗粒 ;脑内接种WNV可致全部乳鼠死亡 ;灭活病毒可诱导小鼠产生WNV抗体 ;经RT PCR扩增 ,在WNV培养液及感染乳鼠脑组织中均检测到目的基因片段 ,且仅与WNV有较高的同源性 (94 %～ 1 0 0 %)。结论　本研究使用的WNV毒株可供进一步开展WNV的相关研究。Objective To investigate the biological characteristics of an imported West Nile virus (WNV) strain, covering morphology, pathogenicity, cell line sensitivity. Methods Cytopathogenic effect (CPE) caused by WNV was observed in Vero-E6 and C6/36 cells, and the negative stain and tissue slice samples for electron microscopy were prepared. Suckling mice were inoculated intracerebrally to observe the pathogenicity of WNV. BALB/c mice were immunized with inactivated WNV, and their serum antibodies against WNV were measured by indirect immunofluorescence assay (IFA). A RT-PCR method was used to detect viral RNA, and then the homology analysis of nucleoside sequence was performed with BLAST program. Results WNV-caused CPEs were characterized by cell shrinking and fusion in Vero-E6 and C6/36 cells respectively. The morphologic observation showed that the virion was spheral particle (approximately 30-50 nm in diameter) with envelope. Suckling mice inoculated with WNV were all killed. Serum antibodies against the virus were detectable in BALB/c mice immunized with inactivated WNV. The target DNA fragments which were only matched with WNV could be amplified from both the virus cultures and brain tissues of infected suckling mice. Conclusion The results can be used for further WNV-related studies.

关 键 词：生物学特征 Vero-E6 病毒株 BALB/c小鼠 间接免疫荧光试验 RT-PCR检测 RT-PCR扩增 C6/36细胞 细胞病变效应 WNV 相关研究 脑内接种 西尼罗病毒 细胞敏感性 生物学性状 形态学鉴定 IgG抗体 同源性比较 核苷酸序列 免疫原性 

分 类 号：R373[医药卫生—病原生物学]