登革病毒感染性转录体技术新途径的初步研究

发布时间：2018-03-29 08:37

本文选题：登革病毒　切入点：RT-PCR技术　出处：《第一军医大学》2006年硕士论文

【摘要】：一、研究背景和目的登革病毒(Dengue virus，DEN)为黄病毒属成员，是引起登革热(DF)和登革出血热(DHF)／登革休克综合症(DSS)的病原体，有4种不同的血清型(1-4型)。主要传播媒介是埃及伊蚊和白蚊伊蚊。流行于热带和亚热带的100多个国家和地区，我国自1978年以来主要在海南、广东、广西、台湾和福建等东南沿海地区发生。世界卫生组织估计全球每年DF发病人数约5千万到1亿，并导致25至50万DHF病例和2.4万人死亡。DF已成为全球范围内严重的公共卫生问题，因此加强对登革病毒致病机理及疫苗研究具有重要的现实意义。登革病毒属黄病毒属(Flavivirus)，是一类有包膜的单股、正链RNA病毒，在复制过程中不形成DNA中间体，因此要在基因水平研究其特征存在一定的困难，因为突变、重组、克隆等分子生物学技术都是在DNA水平上的操作。感染性转录体技术提供了解决这一难题的新途径。该技术是把病毒RNA逆转录成cDNA，并置于RNA聚合酶启动子的下游，在RNA聚合酶的作用下体外转录出全长正链RNA，，经转染敏感细胞后复制出子代病毒颗粒。由于这种子代病毒来自cDNA，研究者就可以在DNA水平上进行操作，从而达到人为改造病毒，研究其复制、表达与致病机理的目的。本研究选择登革2型病毒(Dengue 2 virus，DEN2)NGC株进行感染性转
[Abstract]:I. background and purpose of the study. Dengue virus (DEN), a member of the genus Flavovirus, is the cause of dengue fever (DFD) and dengue haemorrhagic fever (DHFF / DSS). There are four different serotypes of Aedes aegypti and Aedes albopictus. Aedes aegypti and Aedes albopictus are prevalent in more than 100 countries and regions in the tropics and subtropics. China has mainly been in Hainan, Guangdong, Guangxi since 1978. Southeast coastal areas such as Taiwan and Fujian. The World Health Organization estimates that about 50 to 100 million people worldwide suffer from DF every year, causing 25 to 500000 DHF cases and 24000 deaths. DF has become a serious public health problem worldwide. Therefore, it is of great practical significance to strengthen the research on the pathogenesis and vaccine of dengue virus. Dengue virus belongs to Flavivirus, a class of single-stranded, positive stranded RNA virus with envelope, which does not form DNA intermediates during replication, so it is difficult to study its characteristics at the gene level because of mutation and recombination. Molecular biological techniques such as cloning are operated at the DNA level. Infectious transcriptome techniques provide a new way to solve this problem. This technique is to reverse the viral RNA into cDNAs and place them downstream of the RNA polymerase promoter. Under the action of RNA polymerase, full-length positive RNAs are transcribed in vitro, and then transferred into sensitive cells to replicate the offspring virus particles. Because the progeny virus comes from DNA, the researchers can manipulate it at the DNA level, so that the virus can be artificially modified. To study its replication, expression and pathogenesis. In this study, we selected dengue 2 virus Dengue 2 virus DEN2NGC strain for infectious transformation.
【学位授予单位】：第一军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R373

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