摘要
目的对湖州地区一起急性腹泻疫情中检出的GI.8型诺如病毒进行序列测定和分子特征分析。方法根据GI型诺如病毒保守序列自行设计5对引物,用一步法RT-PCR分段扩增GI.8型诺如病毒CHN/Huzhou/N10的全基因组序列,采用生物信息学相关软件对序列进行整理和分析。结果 CHN/Huzhou/N10全长7 740bp,编码区包括3个开放读码框架(ORF1~ORF3),分别长5 400bp(5~5 404nt)、1 632bp(5 388~7 019nt)和642bp(7 019~7 660nt)。RdRp区,VP1区和VP2区的系统进化分析显示CHN/Huzhou/N10属于GI.8基因型。衣壳蛋白区的氨基酸序列分析表明,与原型株Boxer/2001/US相比,CHN/Huzhou/N10VP1区的氨基酸序列共有16个变异位点,12个位于P2区。其中aa347T→S,aa397T→E这2个变异位点分别位于HBGA受体结合袋位点Ⅱ和Ⅲ内。结论本文获得了我国GI.8型诺如病毒CHN/Huzhou/N10的全基因组序列,相关序列信息可用于病毒的遗传进化研究、快速诊断试剂的研发以及疫苗的设计。
We identified and characterized the full-length genome of a GI.8 norovirus strain CHN/Huzhou/N10 isolated in an outbreak in Huzhou,China.The full-length genome of CHN/Huzhou/N10 was amplified using five pairs of primers which were designed according to the full-length GI norovirus genome sequences published in GenBank database.Multiple alignments were performed using DNAStar,the phylogenetic relationship of CHN/Huzhou/N10 and the representative NoV（Norovirus）strains from each genogroup were assessed using the software MEGA version 6.0.The viral genome of CHN/Huzhou/N10 was 7 740 nucleotides in length,which was consist of three ORFs spanning 5-5 404nt（ORF1）,5 388-7 019nt（ORF2）,and7 019-7 660nt（ORF3）,respectively.Phylogenetic analysis based on polymerase and capsid sequences VP1 and VP2 region indicated that CHN/Huzhou/N10 belonged to GI.8 genotype.The amino acid sequence analysis of the VP1 region showed that CHN/Huzhou/N10 had 16mutations compared with the representative strain Boxer/2001/US,12 of these variations were located in the P2 subregion.Moreover,a single amino acid change（T347S）occurred at histo-blood group antigen（HBGA）binding site Ⅱ and another single amino acid change（T397E）occurred at HBGA binding site Ⅲ.In this study,the first full genome of norovirus GI.8 isolated in Huzhou,China was extensively characterized.The data would be helpful not only for the epidemiology study,but also for the diagnostic tool development and effective vaccine design in the future.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2017年第7期613-616,623,共5页
Chinese Journal of Zoonoses
