诺如病毒RdRp区和核心区部分基因克隆及序列分析

Cloning and phylogenetic analysis for RDRP and capsid partial cds of norovirus strain

目的:明确引起急性胃肠炎暴发的诺如病毒RdRp区和衣壳区部分核酸序列,确定病毒基因亚型。方法:采集腹泻病人粪便标本,提取病毒核酸,通过RT-PCR扩增长度为387 bp的病毒RNA多聚酶区(RdRp)区和核心区部分核酸序列,并克隆至T载体,转化大肠杆菌,筛选阳性菌落,对插入片段测序,进行遗传进化分析。结果:确认此次流行株与韩国Seoul/0559/株和美国Shelby株核酸序列相似性最高。结论:国内首次发现诺如病毒GII/12亚型引起急性胃肠炎暴发。

Objective：To reveal the nucleotide sequence of RdRp and capsid protein of Norovirus which aroused an acute gastroenteritis outbreak and to determine virus genotype.Methods：Nucleic acid of Norovirus was extracted from stool samples.The amplifications were carried out targeting the nucleotide sequences of RdRp and capsid protein which were 387bp in length.Then the RT-PCR products were cloned to T vector and transformed into E.coli and sequences were determined.The phylogenetic analysis was implemented.Results：Nucleotide sequence comparison showed this strain was highly identical to those which were named with Seoul/0559 and Shelby.Conclusion：Acute gastroenteritis outbreak which was aroused by Norovirus GII/12 in Hebei Province was confirmed for the first time.