h3n2亚型猪流感病毒致病性试验及其反向遗传操作系统的构建预防兽医学专业论文.docxVIP

异性扩增，再经过一系列克隆技术，将这些片段分别连接至pHW2000双向 异性扩增，再经过一系列克隆技术，将这些片段分别连接至pHW2000双向 表达载体上，成功构建了具有感染性的重组质粒。然后通过细胞转染技术， 将重组质粒转染到293T和MDCK共培养细胞中，以期拯救出rgNNXD和 rgNNXD-JG 1／NS。 关键词：猪流感病毒致病性反向遗传学病毒拯救嵌合病毒 万方数据 PATHoGENICITY PATHoGENICITY EXPERIMENT oF H3N2 SUBTYPE SWINE INFLUENZAⅥRUS AND THE CONSTRUCTION oF REVERSE GENETIC SYSTEM ABSTRACT As we know，pigs can be infected by both the human and avian influenza vires，beacuse there are two kinds of sialic acid receptor in pig’s upper respiratory tract(SAa-2，3-Gal and SAa-2，6-Gal)．This is the reason why pigs are known as“mixing vessels”．The H IN 1 swine influenza virus outbreak in 2009 causing a human influenza pandemic and was introduced into pig herd soon afterwords．So far，there are various types of recombinant viruses exist in herd． It is of great significant for public health that control and prevention of SW outbreak by intensifying the monitoring and understanding it’S biological characteristics． In order to clarify the pathogenicity of a novel recombinant H3N2 subtype SIV to pigs，we used two H3N2 subtype swine influenza virus strains for research，which namely A／Swine／Guangxi／NNXD／20 1 3(H3N2)and A／Swine／ Guangxi／JGl／2013(H3N2)．They all carrying the intemal genes(PB2／PBl／PA／ NP／M)of pdm09 and the surface genes(HA and NA)of seasonal influenza virus． Use these two strains to challenge 1 0 piglets by intratracheal inoculation，each group of 4，and 2 as contr01．All the nasal swab are positive of influenza virus， III 万方数据 万方数据 万方数据 which which revealed the challenged pigs can detox through the nasal cavity．In 3 dpi and 5 dpi，both the JG 1 and NNXD strains can replicate well in lung tissues， among which the JG 1 group has a higher virus titer in lung(reached to 1 04 PFU／mL、)and lung lavage(reached to 1 04·91PFU／mL)．The analysis of histopathology found the alveolar damage and inflammatory are more serious in JGl group．It proved that JGl is more pathogenic than NNXD in pigs． To explore the role of NS gene in swine pathogenic difference，the