云南中缅边境登革1型病毒全基因组序列特征研究

发布时间：2018-11-17 08:44

【摘要】：目的阐明云南省中缅边境地区2013-2015年14株登革1型病毒(DENV-1)全基因组序列特征。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-1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。结果从登革热患者血清中分离到14株DENV-1,其中瑞丽市9株,临沧市3株,昆明市2株。经RT-PCR和序列测定,获得这14株DENV-1的全基因组序列(10 735nt),其开放读码框(95-10 271)编码3 392个氨基酸。系统进化和同源性分析表明,13株为基因I型(G-I),其中瑞丽和临沧本地病例7株,缅甸输入性病例6株;1株为G-III(昆明的印度输入性病例)。云南13株G-I可分为2个进化群,但均与缅甸、泰国等东南亚流行株具有较近的亲缘关系。云南13株G-I的E基因的核苷酸和氨基酸同源性分别为97.02%-100%和98.78%-100%,它们与6株东南亚G-I参考株的核苷酸和氨基酸同源性分别为96.53%-99.53%和97.33%-100%,与DENV-1原型株US_Hawaii核苷酸和氨基酸同源性分别为93.76%-94.45%和95.86%-96.91%。所有云南株和东南亚参考株与US_Hawaii株在结构蛋白或非结构蛋白的氨基酸位点分别存在44和150个位点差异。结论云南中缅边境地区2013-2015年流行的DENV-1均为G-I,并具有基因多样性特点但均为来自缅甸的多个传播来源。
[Abstract]:Objective to elucidate the genomic sequence characteristics of 14 dengue type 1 virus (DENV-1) strains from 2013 to 2015 in Yunnan Province. Methods the virus was isolated by C6 / 36 cell culture method. The whole genome sequence of newly isolated DENV-1 was amplified by RT-PCR. The nucleotide sequence, amino acid sequence homology and phylogenetic analysis were analyzed by bioinformatics software such as ClastalX1.83 and MEGA6. Results 14 strains of DENV-1, were isolated from serum of dengue fever patients, including 9 strains in Ruili city, 3 strains in Linchang city and 2 strains in Kunming city. The whole genome sequence (10 735nt) of the 14 DENV-1 strains was obtained by RT-PCR and sequencing. The open reading frame (95-10 271) encodes 3 392 amino acids. Phylogeny and homology analysis showed that 13 strains were genotype I (G-I), among which 7 were local cases of Ruili and Lincang, 6 were imported from Myanmar, and 1 was G-III (Indian imported case from Kunming). The 13 G-I strains in Yunnan can be divided into two groups, but they are closely related to the epidemic strains in Southeast Asia, such as Burma and Thailand. The nucleotide and amino acid homology of G-I gene were 97.02-100% and 98.78-100%, respectively. The homology of nucleotide and amino acid between them and 6 G-I reference strains in Southeast Asia were 96.53-99.53% and 97.33-100%, respectively. The nucleotide and amino acid homology of US_Hawaii was 93.76% -94.45% and 95.86% -96.91% respectively. There were 44 and 150 amino acid loci of structural or non-structural proteins between Yunnan and Southeast Asia reference strains and US_Hawaii strains, respectively. Conclusion the prevalence of DENV-1 from 2013 to 2015 in the border areas of Yunnan Province is G-I, with the characteristics of genetic diversity, but all of them are multiple transmission sources from Myanmar.
【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;临沧市疾病预防控制中心;昆明市第三人民医院;
【基金】：国家十三、五重大专项(No.2016YFC1200100) 全军青年培育项目(15QNP035) 中国博士后基金(No.2015M582907、2016T91009)联合资助~~
【分类号】：R373

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