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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 isolate murine norovirus (MNV) strains in vitro and identify them. Methods MNV screening was performed by reverse transcription-polymerase chain reaction (RT-PCR) in mice. The detected contents of caecum in MNV-positive mice were inoculated into RAW264.7 cells in murine macrophage cell line, After culturing for a period of time, the sequencing was amplified by RT-PCR. Results MNV was successfully isolated from the passaged RAW264.7 cells and identified. The preliminary analysis of the RT-PCR amplified sequence showed the highest homology with MNV4 S18 strain in Korea (99%). Conclusions The successful replication of MNV in vitro provides the basis for the study of biological characteristics, pathogenesis and diagnostic reagents of MNV. At the same time, as an ideal model of human Norovirus, MNV is also of great significance for the study of human Norovirus .