登革3型病毒义乌流行株免疫原性研究(英文)  

Biological characteristics of dengue virus type 3 isolated from Yiwu,China

作　　者：顾雯[1] 金聪[1] 张硕[1] 张全福[1] 李建东[1] 杭小同[1] 王芹[1] 李川[1] 梁米芳[1] 李德新[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所/病毒基因工程国家重点实验室,北京102206

出　　处：《中国人兽共患病学报》2012年第7期695-699,共5页Chinese Journal of Zoonoses

基　　金：supported by the National High Technology Research and Development Program/863 Program(No.2006AA02A223)~~

摘　　要：目的对2009年浙江省义乌市暴发流行的登革3型病毒进行系统进化分析并初步研究其E蛋白的免疫原性。方法 RT-PCR扩增义乌分离株全基因组,利用MEGA 4分别构建E及NS1区域核苷酸及氨基酸系统发生树,进行系统进化分析。构建重组质粒JESS-ywD3prM/E397JEV-pcDNA5,IFA及Western Blot检测E蛋白在293T细胞中的表达及分泌情况。将纯化后的分泌性E蛋白免疫BALB/c小鼠,通过ELISA、中和试验等方法对体液免疫进行测定。结果登革3型病毒义乌分离株与广州GZ1D3株及印度GWL-25株核苷酸及氨基酸序列同源性均达到99%以上;其分泌性E蛋白可以刺激小鼠产生登革特异性IgG抗体及针对同型病毒的中和抗体。结论义乌分离株属于登革3型病毒亚型Ⅲ,其分泌性E蛋白具有一定的免疫原性。This study performed phylogenic analysis on a dengue strain isolated from an outbreak of dengue fever in 2009 at Yiwu City of Zhejiang Province, China, and further to analyze the immunogenicity of E protein of this viral isolate. Firstly, the viral genome was amplified by RT-PCR and phylogenetic trees were constructed by MEGA 4 based on both nucleotide and amino acid sequences of E and NS1 proteins. The phylogenetic analysis showed that the similarity of Yiwu strain with the Guangzhou GZID3 strain and the India GWL-25 strain was over 990/00. Secondly, the expression plasmid of E protein was con- structed and transfected into 293T cells. The secreted E protein were then purified by sucrose density gradient centrifugation and used to inoculate BALB/c mice. The humoral immunity was evaluated by ELISA and neutralizing antibody analysis. Re- sults showed that the E protein of Yiwu strain could induce dengue specific IgG antibodies and neutralizing antibodies. There- fore, the study found that the Yiwu strain was classified into the subtype III of dengue virus type 3 （DENV-3）, and the E pro tein of this strain had strong immunogenicity

关 键 词：登革3型病毒 义乌分离株 生物学特性 

分 类 号：R373[医药卫生—病原生物学]