1株野生鼠诺如病毒的全基因组序列测定及分析

摘要

目的 为确定1株通过高通量测序获得的GV型野生鼠诺如病毒(MNV)变异株的基因组序列特征.方法 将在山东地区采集的210只鼠经解剖后收集的肠道内容物和肝脏进行核酸提取,质检合格后送于华大基因进行高通量测序,获得一株GV型野生鼠诺如病毒.设计特异性巢式引物利用RT-PCR方法扩增该病毒全基因组.利用DNAMAN,MEGA5.0软件进行序列比对及系统进化分析.结果 获得的GV型野生MNV变异株全基因组长度7 382 bp,与从数据库中选取的30株参考毒株全基因组序列相似度为76.8％ ～78.1％,其中VP1中的高变区P2编码的氨基酸相似度仅为60.9％ ～78.1％,表明该株MNV具有较高的变异率.结论 本研究分离出了一株野生鼠诺如病毒,丰富了国内MNV的基因库,其变异特征为了解国内MNV的分子遗传特征及进化来源提供了参考依据.%Objective To identify the molecular structure and genome homology characteristics of a GV murine norovirus (MNV) from wild mice.Methods Nucleic acids were extracted from collected mouse intestinal specimens and subjected to high-throughput sequencing.The whole genome sequence was obtained by RT-PCR and RACE method.Results The MNV genome was 7,832 bp in length,with 76.8％-78.1％ nucleotide identity to other selected murine norovirus strains in GenBank.The P2 region,the hypervariable area in viral protein 1 gene shows highest heterology,and the identity was among 60.9％-78.1％,lower than other genes regions.The result demonstrated the higher variability of this newly isolated WNV strain.Conclusions The wild-type MNV isolated in this study is first reported in China,which enriched the domestic MNV gene pool and provided a reference for understanding the molecular genetic characteristics and evolutionary origin of MNV in China.