H1N1甲型流感—猪瘟病毒双价活载体疫苗研制

发布时间：2018-05-03 13:29

本文选题：H1N1 + 甲型流感　；参考：《山东师范大学》2015年硕士论文

【摘要】：猪瘟（Classical swine fever CSF），又名猪霍乱或欧洲猪瘟，是一种高致病性，高致死率和高传染性的多系统出血性疾病。猪瘟病毒为其病原，该病的天然宿主为家猪和野猪，历史上有过多次的猪瘟的大爆发，每一次的爆发都会造成巨大的经济损失，对各国的家猪养殖业和国际间的猪产品贸易危害极大，被世界动物卫生组织（OIE）列为A类传染病。猪瘟病毒属于黄病毒科（Flaviviridae family），瘟病毒属（Pestivirusgenus）。猪瘟病毒是单股正链RNA病毒，病毒颗粒的直径大小为40 60nm，基因组大小为12.3kb。其外则具有包膜结构。猪瘟病毒的基因组主要包括一个5’端非编码区（5’UTR），其后面是一个巨大的开放阅读框（ORF）区，最后面则是一个3’端非编码区（3’UTR）。猪瘟病毒有4个结构蛋白：衣壳蛋白C，囊膜蛋白Erns, E1和E2。8个非结构蛋白：Npro, P7, NS2, NS3,NS4A, NS4B, NS5A和NS5B。目前我国预防猪瘟病毒所用的疫苗主要为1954年选育出“54－III系兔化毒”，即C株。C株以其免疫力强，保持效果持久，遗传性质稳定及无残余毒力等优点，在国际上被公认为效果最好的猪瘟弱毒疫苗，已经得到大面积推广和应用，在开发作为其他猪病疫苗载体方面也有很好的前景。流行性感冒病毒（influenza virus），属于正粘病毒科（Orthomyxoviridae）流感病毒分为甲（A）、乙（B）、丙（C）三型，是流行性感冒（流感）的病原体。上个世纪共发生过三次世界性的大流行：1918年，H1N1引起的西班牙流感，1957年H2N2引起的亚洲流感和1968年，H3N2引起的香港流感。进入到本世纪，2009年4月，一种新型的H1N1猪流感病毒在人类身上检测出来，并且于2009年9月份开始演变为世界性的大流感在人类中迅速传播。目前，全世界大流行的H1N1甲型流感正在威胁人类健康，对其病原分析发现该病毒与猪流感病毒有最高的核酸序列同源性，表明其起源于猪流感病毒。在我们投入大量的人力物力防控该病毒在人与人之间流行的时候，如何从源头上有效切断流感病毒从猪传染给人是必须考虑的问题。对于病毒性疾病，最有效的防控办法是用有效的疫苗预防接种，而目前我国尚没有预防猪的H1N1甲型流感的正规疫苗可用。因此，研发控制H1N1甲型流感病毒在猪与猪之间传播的疫苗迫在眉睫。本研究利用我们实验室已经构建的兔化弱毒株感染性cDNA以及从合作单位获得的H1N1的HA基因，构建猪瘟-H1N1甲型流感嵌合疫苗，以期在进行猪瘟计划免疫的同时，防控猪流感病毒，避免猪流感病毒在猪与猪之间传播，从而可能从源头上切断H1N1甲型流感病毒从猪传染给人的途径，为H1N1甲型流感病毒的防控奠定基础。同时，本研究建立的CSFV的实验室检测方法灵敏度非常高，，能够准确的检测到PK-15细胞中CSFV的存在，为今后的以兔化弱毒株cDNA为载体的疫苗实验奠定了基础。
[Abstract]:Classical swine fever (CSF) is a multisystem hemorrhagic disease with high pathogenicity, high mortality and high infectivity. Swine fever virus is the pathogen, the natural host of the disease is domestic pigs and wild boars. There have been many outbreaks of swine fever in history, each outbreak will cause huge economic losses. It is very harmful to domestic pig breeding and international trade in pig products, and is classified as Class A infectious disease by the World Organization of Animal Health (OIE). Swine fever virus belongs to Flaviviridae family of Flaviviridae and Pestivirusgenusus. Classical swine fever virus (CSFV) is a single-stranded RNA virus with a particle diameter of 40 ~ 60nm and a genome size of 12.3 kb. On the other hand, it has a envelope structure. The genome of CSFV mainly consists of a 5'-terminal non-coding region, followed by a large open reading frame (ORF) region, and a 3'terminal non-coding region (UTR3). There are four structural proteins of CSFV: capsid protein C, envelope protein Erns, E1 and E2.8 non-structural proteins: npro. P7, NS2, NS3NS4A, NS4B, NS5A and NS5B. At present, the main vaccine used to prevent swine fever virus in China is the "54-III strain rabbit chemical virus" bred in 1954, that is, strain C has the advantages of strong immunity, lasting effect, stable genetic property and no residual virulence. The attenuated swine fever vaccine, which has been recognized as the best vaccine in the world, has been widely used and has a good prospect in the development of other swine vaccine vectors. Influenza virus, Orthomyxoviridae3, is the pathogen of influenza. There were three worldwide pandemics in the last century: Spanish influenza caused by H1N1 in 1918, Asian influenza caused by H2N2 in 1957 and Hong Kong influenza caused by H3N2 in 1968. In this century, in April 2009, a new type of H1N1 swine flu virus was detected in humans, and in September 2009, it began to develop into a worldwide pandemic influenza spread rapidly in humans. At present, pandemic influenza A (H1N1) in the world is threatening human health. It is found that the virus has the highest nucleic acid sequence homology with swine influenza virus, which indicates that it originated from swine flu virus. When we invest a lot of manpower and material resources to prevent and control the virus from people to people, how to effectively cut off the transmission of influenza virus from pigs to people is a problem that must be considered. For viral diseases, the most effective method of prevention and control is to use effective vaccination, but at present, there is no regular vaccine available for the prevention of swine H1N1 influenza A in China. Therefore, it is urgent to develop a vaccine to control the transmission of H1N1 influenza A virus between pigs and pigs. In this study, we used the HA gene of rabbitized attenuated strain cDNA and H1N1 obtained from our laboratory to construct swine fever H1N1 influenza A chimeric vaccine, in order to prevent and control swine flu virus while carrying out swine fever program immunization. To avoid the transmission of swine flu virus between pigs and pigs, it may cut off the transmission of H1N1 A influenza virus from swine to human from the source, and lay the foundation for the prevention and control of H1N1 influenza A virus. At the same time, the laboratory detection method of CSFV established in this study has very high sensitivity and can accurately detect the presence of CSFV in PK-15 cells, which will lay a foundation for the future vaccine experiment with rabbit attenuated strain cDNA as vector.
【学位授予单位】：山东师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S859.797

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