云南省登革4型病毒全基因组序列特征研究

发布时间：2018-03-23 21:15

本文选题：登革型病毒　切入点：全基因组　出处：《中国人兽共患病学报》2017年10期

【摘要】：目的阐明云南省2015年5株登革4型病毒(DENV-4)流行株的全基因组序列特征及分子流行病学特点。方法采用C6/36细胞培养法分离病毒,用RT-PCR法扩增新分离DENV-4的全基因组序列,采用ClastalX1.83和MEGA6等生物信息学软件进行核苷酸和推导氨基酸序列同源性及系统进化分析。结果从2015年云南省瑞丽市登革热患者血清中分离到5株DENV-4(本地病例2株,来自缅甸腊戍和南坎市输入性病例3株)。经RT-PCR和序列测定,获得这5株DENV-4的全基因组序列(10 661nt),其开放读码框(103-10 264)编码3 386个氨基酸。全基因组或结构蛋白和非结构蛋白基因序列的系统进化和同源性分析表明,云南分离株间高度同源并聚集为一个进化支,并与泰国不同年代DENV-4基因I型(G-I)流行株具有较近的进化关系和较高的同源性,同属G-I。云南株和泰国株均与同基因型的DENV-4原型株(H241,1956年菲律宾)和中国广州1990年B5株亲缘性和同源性都较低。云南株与H241株在结构蛋白或非结构蛋白中分别存在21和45个氨基酸位点差异。结论首次获得云南省DENV-4分离株的全基因组序列并发现它们与近期东南亚DENV-4G-I流行株亲缘关系较近。首次证实云南省存在DENV-4本地流行,传播来源为相邻缅甸北部边境地区。云南分离株某些氨基酸位点的改变是否与其抗原性和毒力有关尚需进一步研究。
[Abstract]:Objective to elucidate the characteristics of the whole genome sequence and molecular epidemiology of five dengue type 4 virus (DENV-4) strains in Yunnan Province in 2015. Methods the virus was isolated by C6 / 36 cell culture method and the whole genome sequence of newly isolated DENV-4 was amplified by RT-PCR. Nucleotide and deduced amino acid sequence homology and phylogenetic analysis were carried out using bioinformatics software such as ClastalX1.83 and MEGA6. Results five DENV-4 strains were isolated from serum of dengue fever patients in Ruili City, Yunnan Province in 2015. Three strains of imported cases from Lasso and Namkam, Myanmar. RT-PCR and sequence analysis were performed. The whole genome sequence of these five DENV-4 strains was 10661ntTN, which encodes 3 386 amino acids. The phylogeny and homology analysis of the whole genome or structural protein and non-structural protein gene sequence showed that the sequence of the whole genome or structural protein and the non-structural protein gene were homologous. The isolates from Yunnan were highly homologous and clustered into an evolutionary branch, and had a close evolutionary relationship and high homology with the epidemic strains of DENV-4 gene type I in Thailand in different ages. The relationship and homology between Yunnan strain and H241 strain were lower than those of H241 (1956, Philippines) and 1990 B5 strain (Guangzhou, China). There were 21% structural protein or non-structural protein in Yunnan strain and H241 strain, respectively. Conclusion the whole genome sequence of DENV-4 isolates from Yunnan Province was obtained for the first time, and their genetic relationship with the recent DENV-4G-I epidemic strains in Southeast Asia was found to be close. It is the first time to confirm the existence of DENV-4 endemic in Yunnan Province. The source of transmission is the border area of northern Myanmar. Whether the change of amino acid sites of Yunnan isolate is related to its antigenicity and virulence should be further studied.
【作者单位】：成都军区疾病预防控制中心;瑞丽市疾病预防控制中心;
【基金】：国家十三五重大专项(No.2016YFC1200100) 全军青年培育项目(No.15QNP035) 中国博士后基金项目(No.2015M582907和2016T91009)联合资助~~
【分类号】：R373

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