2018年湖南省登革病毒2型分离株非结构蛋白1基因的鉴定及分析  

Identification and analysis of nonstructural protein 1 gene of dengue virus type 2 isolated in Hunan Province,China in 2018

作　　者：冯石军 刘雨 陈珺 段素琴[2] 饶清 陈俊英[2] 管娇琼 FENG Shi-jun;LIU Yu;CHEN Jun;DUAN Su-qin;RAO Qing;CHEN Jun-ying;GUAN Jiao-qiong(Qiyang County People's Hospital Affiliated to Changsha Medical College,Yongzhou 426100,Hunan Province,China;不详)

机构地区：[1]长沙医学院附属祁阳县人民医院,湖南永州426100 [2]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室,云南昆明650031

出　　处：《中国生物制品学杂志》2020年第12期1357-1362,共6页Chinese Journal of Biologicals

基　　金：云南省自然科学基金(2016FA029)。

摘　　要：目的鉴定并分析2018年湖南省登革病毒2型(dengue virus 2,DENV-2)分离株的非结构蛋白1(non-structural protein 1,NS1)基因。方法利用试剂盒提取登革热患者血清RNA,RT-PCR法鉴定病毒类型及扩增11株分离株NS1基因,并进行基因测序。应用MEGA 7.0软件分析分离株与DENV-2标准株(KM204118)的NS1基因核苷酸及氨基酸序列,利用最大似然法(1000 Boot strap replicate)构建NS1基因的系统进化树。结果共获得11株分离株的NS1基因,经比对发现,除分离株HNNS201804和HNNS201810外,其余分离株在NS1基因第585位碱基由T变为C(无义突变);HNNS201806的NS1基因第936位碱基由C变为T(无义突变)。11株分离株NS1基因其他突变位点均相同,共68处发生碱基突变,其中9个为有义突变。11株分离株与China-ZJ(2017-MH110594)、China-GZ(2018-MK564485、2015-KX621245、2017-MK564483)、Thailand(2017-LC410191)、Singapore(2016-MF314189)、Philippines(2015-KU517847)有较近的亲缘关系。结论2018年湖南省11株分离株均为DENV-2,可能是浙江或广州输入株,也可能是泰国、新加坡和菲律宾等东南亚国家的输入株。Objective To identify and analyze the nonstructural protein 1(NS1)gene of dengue virus type 2(DENV-2)isolated in Hunan Province,China in 2018.Methods RNA was extracted from sera of patients with dengue fever by kit,while the virus type was identified by RT-PCR,and the NS1 genes of 11 isolates were amplified and sequenced.The nucleotide and amino acid sequences of NS1 gene of the isolates and DENV-2 standard strain(KM204118)were analyzed by MEGA 7.0 software,and the phylogenetic tree of NS1 gene was constructed by the maximum likelihood method(1000 boot strap replicate).Results The NS1 gene of 11 isolates were obtained.It was found that,except those of isolates HNN201804 and HNNS201810,the bases at site 585 of NS1 gene of the other isolates changed from T to C(nonsense mutation),and that at site 936 of isolated HNNS201806 from C to T(nonsense mutation).However,the other mutation sites of NS1 genes were identical in the 11 isolates,and there were 68 base mutations including 9 meaningful mutations.The 11 isolates were closely related to strains China-ZJ(2017-MH110594),China-GZ(2018-MK564485,2015-KX62-1245,2017-MK564483),Thailand(2017-LC410191),Singapore(2016-MF314189),and Philippines(2015-KU517-847).Conclusion All the 11 strains isolated in Hunan Province in 2018 were DENV-2,which might be imported from Zhejiang or Guangzhou,or from Thailand,Singapore,Philippines and other Southeast Asian countries.

关 键 词：登革病毒2型 NS1基因 基因突变 系统进化树 

分 类 号：R373.33[医药卫生—病原生物学] Q789[医药卫生—基础医学]