摘要
目的体外分离出小鼠诺如病毒(murine norovirus,MNV)毒株,并对其进行鉴定。方法采用逆转录-聚合酶链反应(RT-PCR)法对小鼠进行MNV筛查,将检测出的MNV阳性小鼠的盲肠内容物经过处理接种至鼠源巨噬细胞系RAW264.7细胞,培养一段时间后用RT-PCR法扩增测序。结果成功通过传代RAW264.7细胞分离出MNV,并经过鉴定确认,对RT-PCR扩增序列进行初步分析,与韩国MNV4 S18株同源性最高,达99%。结论 MNV在体外的复制成功,为MNV的生物学特性、致病机制和诊断试剂等方面的研究提供了基础,同时作为人诺如病毒研究的理想模型,对人诺如病毒的研究也有重大意义。
Objective To establish the replication system of murine norovirus（MNV） in vitro.Methods The cecum contents of mice containing MNV were inoculated into RAW264.7 cells.The pathogenesis of inoculated cells was observed and the supernatants of cell culture were collected after 72 hours.The replication of MNV was confirmed by RT-PCR and gene sequencing.Results The replication of MNV was observed in the supernatant and cells of inoculated RAW264.7 cell lines.Cytopathic effect（CPE） was obvious after 72 hours of infection.High nucleotide sequence homology（above 97%） was also observed when the isolated strain was compared with other reported strains.The highest nucleotide sequence homology of 99% was seen between the isolated strain and the strain S18 of MNV 4 isolated from South Korea.Conclusions The in vitro replication system of MNV is established.It can facilitate research for MNV biology,pathogenesis and laboratory diagnostics.
出处
《中国病毒病杂志》
CAS
2011年第4期293-296,共4页
Chinese Journal of Viral Diseases
基金
中国药品生物制品检定所中青年发展研究基金(2010C3)
