本文選題：狂犬病病毒　切入點(diǎn)：G基因　出處：《吉林農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：狂犬病是一種嚴(yán)重影響世界公共衛(wèi)生的人獸共患病。一旦被感染而不采取有效措施,出現(xiàn)癥狀后死亡率幾乎100%。據(jù)世界衛(wèi)生組織報(bào)道,全世界每年因狂犬病導(dǎo)致的死亡人數(shù)為5~6萬人,而絕大多數(shù)疫情出現(xiàn)在發(fā)展中國家,其中主要在亞洲和非洲。我國在世界上屬狂犬病高發(fā)地區(qū),發(fā)病和死亡人數(shù)僅次于印度,居世界第二位�？袢〔《�(rabies virus,RABV)幾乎感染所有溫血?jiǎng)游?是目前發(fā)現(xiàn)宿主感染譜最為廣泛的一種RNA病毒。鑒于狂犬病的巨大危害,狂犬病的預(yù)防控制日益得到關(guān)注。目前,世界各地都采取了一系列的措施來控制狂犬病,但是距離徹底控制狂犬病還需一定的時(shí)間。要想成功控制狂犬病的蔓延態(tài)勢,針對狂犬病的特點(diǎn)進(jìn)行控制才能達(dá)到更好的效果。狂犬病病毒的宿主十分廣泛,幾乎能感染所有的溫血?jiǎng)游?這是其它任何病毒都不具備的特點(diǎn),但是RABV如此廣宿主感染譜的原因目前尚不清楚。包括RABV在內(nèi)的RNA病毒是由一系列基因序列相近而又不完全相同的突變體構(gòu)成,這稱為病毒準(zhǔn)種現(xiàn)象。病毒在動(dòng)物或細(xì)胞內(nèi)連續(xù)傳代后會產(chǎn)生適應(yīng)性,而RABV的準(zhǔn)種在病毒感染譜中及宿主適應(yīng)的過程中是否發(fā)揮作用,尚沒有詳細(xì)的研究。為研究狂犬病病毒街毒株在新宿主中的傳播及其在適應(yīng)新宿主過程中準(zhǔn)種的變化規(guī)律,本研究首先選取處于不同進(jìn)化分支的5株街毒株(HuND03、SXLF03、FJ02、CQQJD09、927A)和固定毒株CVS-11,將采集的感染這6株病毒致死的動(dòng)物腦組織懸液腦內(nèi)接種乳鼠,此為F1代,在乳鼠水平連續(xù)傳至15代。選取乳鼠水平傳代獲得的HuND03、SXLF03、FJ02、CQQJD09、927A的F2、F7、F10、F15代鼠腦和上述毒株的原始腦組織(F0代),分別進(jìn)行各代次的狂犬病病毒G基因全長序列的擴(kuò)增,將擴(kuò)增后的樣品使用Illumina Hiseq 2500測序方法進(jìn)行測序,使用Vivan軟件進(jìn)行多序列比較分析。結(jié)果表明:毒株在適應(yīng)性傳代過程,準(zhǔn)種變異體始終是動(dòng)態(tài)變化的,6株毒株F0、F2、F7、F10、F15代G基因上共有有302個(gè)位點(diǎn)出現(xiàn)突變,突變的類型只發(fā)現(xiàn)了點(diǎn)突變,并沒有發(fā)現(xiàn)缺失突變、插入突變、提前或是終止突變的發(fā)生。出現(xiàn)頻率最高的是A→G,其次是T→C。總體來說,同義突變變異體的數(shù)量大于非同義突變。研究發(fā)現(xiàn)RABV準(zhǔn)種受到了宿主陽性選擇的作用,陽性選擇位點(diǎn)主要分布在G蛋白膜外區(qū),但是變異體的數(shù)量和遭受陽性選擇的位點(diǎn)隨著毒株在乳鼠腦內(nèi)傳代代次的改變而改變。準(zhǔn)種變異體在新宿主體內(nèi)適應(yīng)性傳代過程中表現(xiàn)出3種不同的機(jī)制:1.適應(yīng)新宿主的變異體出現(xiàn)2.不適應(yīng)的變異體被淘汰3.變異體始終處于競爭狀態(tài)。研究發(fā)現(xiàn)RABV序列較為保守,在新宿主(乳鼠)體內(nèi)適應(yīng)性傳代過程中只出現(xiàn)了少數(shù)變異體,但是可能這少數(shù)變異體就足以使RABV完成跨宿主傳播過程。與其他RNA病毒不同,RABV具有廣泛宿主譜的特性并非后天逐步進(jìn)化而來,而是RABV在形成之初,就具備適應(yīng)不同宿主的特性。RABV這種高度保守的特性,可能是其迅速完成適應(yīng)性進(jìn)化的基礎(chǔ)。本研究通過研究RABV準(zhǔn)種,闡明了其在宿主適應(yīng)性過程所表現(xiàn)出的動(dòng)力學(xué)特征,初步了解了狂犬病病毒具有廣泛宿主感染譜的原因,研究結(jié)果對于最終闡明RABV在宿主跨種傳播分子遺傳機(jī)制提供了新的視角,為狂犬病科學(xué)防控奠定了理論基礎(chǔ)。
[Abstract]:Rabies is a serious impact on world public health Amphixenosis. Once infected and do not take effective measures, symptoms of mortality after almost 100%. WHO reported around the world each year the number of deaths caused by rabies caused by 5~6 million people, while the vast majority of the epidemic situation in developing countries, mainly in Asia and Africa. China is a high incidence of rabies regions in the world, the incidence and deaths after India, ranked second in the world. The rabies virus (rabies virus, RABV) infect almost all warm blooded animal, is found in the host spectrum of infection as a RNA virus. In view of the great harm of rabies, rabies prevention and control have been paid more and more attention at present, around the world have taken a series of measures to control rabies, rabies control completely but the distance needed for some time. In order to successfully control The trend of the spread of rabies, according to the characteristics of rabies control in order to achieve better results. The rabies virus host is very wide, almost can infect all warm blooded animal, which is characteristic of any other virus does not have, but RABV is so broad host spectrum of infection reason is unclear. Including RABV, RNA virus by a series of gene sequences are similar but not identical mutants, this is called a virus quasispecies. The virus was subcultured in animal cells or will produce adaptability, while RABV quasispecies infection does play a role in adaptation and host spectrum in the process of the virus, there is no detailed research for communication research. Rabies street virus strains in the host and in the adaptation process quasi variation of new host species, this paper selects 5 strains of street strains in different evolutionary branches (HuND03 SXLF03, FJ02, CQQJD09927A), and a fixed strain CVS-11, the collected brain tissue infected animal of the 6 strains of the virus lethal suspension intracerebral inoculation of suckling mice, the F1 generation in neonatal rat level 15 passages. Select the rat level obtained HuND03 passage SXLF03, FJ02, CQQJD09927A, F2. F7, F10, the original brain tissue F15 generation rat brain and the strain (F0), were amplified the full-length sequence of rabies virus G gene in each generation, the amplified samples using Illumina Hiseq 2500 sequencing methods were sequenced using Vivan software for multi sequence analysis. The results showed that the strains in the adaptability passage, quasi variants is always dynamic, 6 strains F0, F2, F7, F10, F15 G gene has 302 loci mutation, mutation types found only point mutations and found no deletion mutation, insertion mutation, mutation is terminated in advance or . the highest frequency is A to G, followed by T, C. overall, the number of synonymous mutation variants than non synonymous mutations. The study found that the RABV quasispecies is host of positive selection, positive selection sites mainly distributed in the outer membrane protein G, but the number of variants and subjected to positive selection sites with the passage of time changes in the generation of rat brain strain change. Quasispecies variants in the new host adaptive passage process showed 3 different mechanisms: 1. to adapt to the new host to the 2. variants appear variants were eliminated 3. variation of body is always in a competitive state. Research found that RABV sequence is more conservative, in a new host (rat) only a few variants appear in the process of adaptive passage, but perhaps this is enough to make a few variants of RABV to complete the process of inter host transmission. Different from other RNA virus, RABV Is not a characteristic of a broad host spectrum acquired gradually evolved, but RABV at the beginning of the formation, is to adapt to the characteristics of different host characteristics of.RABV this highly conserved, likely is the basis of the rapid completion of adaptive evolution. Through the study on the quasispecies of RABV, illustrates the show in the host adaptive process dynamics characteristics., a preliminary understanding of the rabies virus with a broad host spectrum of infection, the results of the study for clarifying the cross species transmission of RABV in host molecular genetic mechanism provides a new perspective, lays a theoretical foundation for the prevention and control of rabies.

【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S852.65

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