2株猪h9n2亚型流感病毒的分离鉴定及对小鼠致病性的研究-预防兽医学专业毕业论文.docxVIP

药物敏感；M2蛋白上的第31位发生了由S_N的突变，表明这两株毒对 药物敏感；M2蛋白上的第31位发生了由S_N的突变，表明这两株毒对 金刚烷胺类药物产生耐药性；在内部基因中，与致病力有关的基因位点PB2 的627、701位，PBl的622、709位，PA的3 l 5、5 15位，NS的92位以 及NP的319位与参考毒株相比均保守。通过2株分离株分子进化树和同源 性分析显示，HA来源于Y280．1ike亚系，NA来源于G9．1ike亚系，M和 PB2来源于G1．1ike亚系，PBl、PA、NS和NP来源于F98．1ike亚系。 对这2株进行鸡胚半数感染量(EID50)试验得出SW／GX／P2／20 1 1和 SW／GX／P3／2011的其L啦ID5do．1mL分别为5．02以及5．23。小鼠致病性结 果显示，小鼠出现了以肺脏为主的病理变化，病毒能够在肺脏以及鼻甲中 复制，但复制水平有一定差异，上呼吸道的鼻甲的复制能力比下呼吸道的 肺脏高。2个毒株最低均可使小鼠体重下降6％，但并未能致死小鼠，之后 至第8天后逐渐恢复正常体重。 关键词：H9N2亚型猪流感全基因测序遗传进化生物学特性致病性 II 万方数据 ISOLATION ISOLATION AND IDENTIFICATION OF TWO STRAINS OF H9N2 SWINE INFLUENZA VIRUS AND THEIRS f’ATHOGENICITY 0N MICE AB STRACT Nowdays，Swine Influenza have been distributed all over the world．The species barrier of pigs is relatively low,it forms a transmission chain of “humen-swine-avian,if infected two kinds of influenza virus at the same time，it’S easily lead to gene reassortment and produce new influenza virus subtypes．All of these indicate the influenza virus of H9N2 subtype is invading human life and to be a serious threat to public health and safety． The study used RT-PCR and embryonated eggs to discuss biological characteristics and isolated virus，molecular genetics and pathogenicity test on mice of the two strains of H9N2 subtype S1 which isolated from pig farms of Baise Leye county,GuangXi in 20 1 1．According to the BLAST result on NCBI，the two isolated strains determined to the influenza virus of H9N2 subtype that the laboratory name them SW／GX／P2／20 1 1 and SW／GX／P3／20 1 1．The ORF of nucleotide of PB2，PB 1，PA，HA，NP,NA，M and NS respectively are 2280bp，2274bp，2151bp，1683bp，1497bp，1410bp，982bp and III 万方数据 83 83 8bp．Two isolates compare homology with A／Canine／Guangxi／1／20 1 1 and A／Equine／Guangxi／3／20 1 1 that is above 99．O％． Sequences analysis of HA protein in 1 09，1 6 1，1 63，202 and 203 sites were very conservative，the 234th is leucine(Leu)and it had the receptor SAa，6Gal specificity．The cleavage site of HA is-·RSSRGLF--and no multiple amino acid i