發(fā)布時(shí)間：2018-03-22 13:50

  本文選題：輪狀病毒　切入點(diǎn)：交叉保護(hù)　出處：《中國(guó)藥品生物制品檢定所》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：輪狀病毒(Rotavirus, RV)是引起嬰幼兒重癥腹瀉最重要的病原,也是導(dǎo)致發(fā)展中國(guó)家嬰幼兒死亡的主要原因之一。輪狀病毒的治療目前尚無(wú)特效藥,臨床上主要采用補(bǔ)液、維持電解質(zhì)平衡及對(duì)癥治療等方法,只能挽救生命,不能控制病毒的傳播。因此開發(fā)輪狀疫苗成為當(dāng)今WHO疫苗發(fā)展計(jì)劃中的首要任務(wù)。 目前國(guó)內(nèi)外批準(zhǔn)上市的疫苗有三種:人減毒活疫苗(Rotarix)、人-牛五價(jià)重配苗(RotaTeq)和羊減毒活疫苗(LLR)[2-4]。我國(guó)羊三價(jià)減毒活疫苗正在進(jìn)行Ⅰ期臨床,人-牛六價(jià)重配苗也處于研發(fā)階段[5]。由于主要是A組輪狀病毒引起兒童重癥腹瀉,所以這些疫苗的生產(chǎn)用病毒株均為A組。 隨著疫苗的研制與上市,對(duì)于疫苗來說究竟是單價(jià)還是多價(jià)能起到最好的保護(hù)效果?單價(jià)的是哪型好?多價(jià)的需要多少價(jià)才可以最大效果保護(hù)好人類?哪種型別的毒株能最大范圍的與其他毒株進(jìn)行交叉免疫保護(hù)?研究發(fā)現(xiàn)動(dòng)物與人類毒株之間有在自然界發(fā)生重組的可能性。由于目前說法不一,并且缺乏基礎(chǔ)研究,對(duì)疫苗的評(píng)價(jià)也造成困難,所以我們開展這個(gè)課題,以期解決目前的瓶頸問題。本課題采用已上市的兩種種輪狀病毒疫苗—葛蘭素史克公司的人減毒活疫苗(Rotarix)和蘭州生物制品研究所的羊減毒活疫苗(LLR)及輪狀病毒W(wǎng)a株、DS-1株,SA11株及ST-3株作為實(shí)驗(yàn)病毒株,進(jìn)行了輪狀病毒免疫交叉反應(yīng)的研究。 本研究就三個(gè)方面進(jìn)行探討,第一部分為輪狀病毒減毒交叉免疫反應(yīng)的研究,以期找到LLR和Rotarix免疫后抗體能最大范圍保護(hù)哪種輪狀病毒的感染。這些為以后輪狀病毒疫苗的研發(fā)與質(zhì)控打下基礎(chǔ);第二部分為從噬菌體展示隨機(jī)十二肽庫(kù)中篩選輪狀病毒特異性結(jié)合肽的實(shí)驗(yàn),挑選出于用輪狀病毒免疫后血清結(jié)合力強(qiáng)的克隆,將獲得的噬菌體克隆進(jìn)行DNA測(cè)序,根據(jù)噬菌體克隆基因中所插入的外源DNA序列推導(dǎo)出呈現(xiàn)的外源多肽氨基酸序列,應(yīng)用DNAStar軟件及其它生物信息網(wǎng)絡(luò)軟件如BLAST軟件將多肽序列比較分析,進(jìn)行蛋白結(jié)構(gòu)數(shù)據(jù)庫(kù)和文獻(xiàn)數(shù)據(jù)庫(kù)檢索;第三部分為輪狀病毒結(jié)構(gòu)蛋白VP5、VP6、VP7、VP8的原核表達(dá)與純化,以期獲得輪狀病毒融合蛋白,并對(duì)其免疫原性進(jìn)行研究。
[Abstract]:Rotavirus (Rotavirus, RV) is the cause of severe diarrhea in infants and young children the most important pathogen, is one of the main causes of infant death in developing countries. The treatment of rotavirus is currently no cure, the main clinical use of rehydration, electrolyte balance and symptomatic treatment methods can save lives, communication can not control the virus. Therefore the development of rotavirus vaccine has become a primary task in the WHO vaccine development plans.
There are three kinds of domestic licensed vaccines, live attenuated vaccine (Rotarix), human bovine reassortant vaccine quinquevalent (RotaTeq) and sheep live attenuated vaccine (LLR) [2-4]. China sheep trivalent live attenuated vaccine ongoing phase I clinical, human bovine reassortant vaccine in six price the development stage is mainly due to [5. Group A rotavirus cause severe diarrhea, so these vaccine virus strains were produced by A group.
With the development and market of the vaccine, a vaccine for what is the price or multivalent can play a protective effect of the best price? What type? How can the price of polyvalent maximum effect to protect human beings? What types of strains can cross immune protection and the maximum range of other strains in research found? The possibility of recombination occurs in nature between animal and human strains. Due to the inconsistency, and lack of basic research, evaluation of the vaccine is difficult, so we carry out this task, in order to solve the bottleneck problem at present. This paper has listed two kinds of rotavirus vaccine, live attenuated human GlaxoSmithKline Co the vaccine (Rotarix) and the Lanzhou Institute of biological products of sheep live attenuated vaccine (LLR) and rotavirus Wa strain, DS-1 strain, SA11 strain and ST-3 strain as the experimental strains of rotavirus disease Study on the cross reaction of toxic immune system.
This study is to explore three aspects, the first part is the study of rotavirus attenuated cross immune reaction, in order to find the LLR and Rotarix antibody to the largest extent to protect what kind of rotavirus infection. The development and quality control after rotavirus vaccine foundation; the second part is to demonstrate the experiment screening of rotavirus specific binding peptides from phage random twelve peptide library, selected for cloning with rotavirus serum adhesion, the phage clones obtained by DNA sequencing, based on the insertion of exogenous DNA gene in phage clone sequence deduced amino acid sequence of exogenous peptides presented, using DNAStar software and other biological information network software such as BLAST software to comparative analysis of the peptide sequence, protein structure database and document database retrieval; the third part is the rotavirus structural protein VP5 The prokaryotic expression and purification of VP6, VP7 and VP8 were expressed in order to obtain rotavirus fusion protein and to study the immunogenicity of rotavirus.

【學(xué)位授予單位】：中國(guó)藥品生物制品檢定所
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R392.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 鄭麗舒;輪狀病毒疫苗發(fā)展及現(xiàn)狀[J];國(guó)外醫(yī)學(xué).病毒學(xué)分冊(cè);2003年01期

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本文編號(hào)：1648930