登革4型病毒Ban18原株及其减毒株的分子特征研究  

Study on the molecular characteristics of two dengue type 4 virus strains,Ban18 strain and its attenuated strain Ban18-30

作　　者：刘志文[1] 俞永新[1] 贾丽丽[1] 董关木[1] 

机构地区：[1]中国药品生物制品检定所疫苗一室,北京100050

出　　处：《中国病原生物学杂志》2008年第5期343-347,389,共6页Journal of Pathogen Biology

摘　　要：目的从分子水平上探讨登革4型病毒Ban18-30减毒株的毒力相关位点和减毒机制。方法Ban18原株及Ban18-30减毒株在Vero细胞中培养增殖,收集病毒液,经RNA抽提后RT-PCR扩增C、prM、E、NS1和3′、5′端核苷酸片段并测序,比较分析结果,研究两株病毒的分子特征。结果RT-PCR成功扩增出目的片段,对两株病毒部分基因测定序列后进行对比分析,两株病毒间在C111位和E155位各有1个氨基酸残基发生变化,在3′UTR的219位发生核苷酸改变。结论从分子水平证明两株病毒确为DEN-4型病毒,某些基因位点的变化与Ban18-30株病毒毒力减弱有关,其中E155位的突变可能关系最大。Objective To explore the potential virulence gene sites of the attenuated Ban18-30 strain and the attenuation mechanism in the molecular level. Methods The Ban18 strain virus and its attenuated strain Ban18-30 virus were cultured in Vero cells and the suspensions of them were harvested. After RNA was extracted, the C, prM, E, NS1, 3＇UTR and 5＇UTR gene were amplified by RT-PCR and then sequenced. The molecular characteristics of the two viruses were analyzed. Results The interest genes were successfully amplified by RT-PCR. The alignment of partial sequences and analysis of the two strains showed that there was an amino acid change each in the C111 and E155 site and a nucleotide acid change in 3＇UTR. Conclusion The two strains are proved to be dengue type 4 virus in the molecular level. Some gene sites mutations may lead to the virulence attenuation of the Ban18-30 virus, in which the most relevant site was E155.

关 键 词：登革4型病毒 基因分析 毒力位点 系统发生树 

分 类 号：R373.33[医药卫生—病原生物学]