吉林省猪流感病毒的分离鉴定及分离株HA、NA基因序列分析

发布时间：2018-06-05 03:17

本文选题：猪流感病毒 + H1N1　；参考：《吉林农业大学》2015年硕士论文

【摘要】：本研究对吉林地区采集到的猪血清样本7909份、鼻拭子及疑似发病死亡猪只的肺脏、支气管等病料5154份,总计13063份样本进行了病毒分离,并且对分离到的毒株进行亚型鉴定。结果显示分离到的三株猪流感病毒均为H1N1亚型猪流感病毒,根据流感病毒命名标准体系分别命名为A/swine/Jilin/1-1/2011、A/swine/Jilin/1-2/2011和A/swine/Jilin/1-3/2011。对分离株HA、NA基因进行克隆和遗传进化分析。基因测序结果表明,SIV分离株A/swine/Jilin/1-1/2011(H1N1)的HA基因序列全长为1638个核苷酸,编码区为40-1636bp,编码532个氨基酸。NA基因序列全长为1420个核苷酸,编码区为4-1413bp,编码470个氨基酸;分离株A/swine/Jilin/1-2/2011(H1N1)的HA基因序列全长为1703个核苷酸,编码区为26-1556bp,编码510个氨基酸。NA基因序列全长为1404个核苷酸,编码区为30-1397bp,编码456个氨基酸;分离株A/swine/Jilin/1-3/2011(H1N1)的HA基因序列全长为1745个核苷酸,编码区为17-1722bp,编码568个氨基酸。NA基因序列全长为1458个核苷酸,编码区为21-1430bp,编码470个氨基酸。裂解位点显示三株均为低致病性毒株,糖基化位点均未发生变异相对保守,受体结合位点显示分离株具有感染禽源的潜力,NA基因耐药性位点显示这三株猪流感病毒对目前使用的神经氨酸酶抑制剂类药物均有效。遗传进化分析结果显示A/swine/Jilin/1-1/2011和A/swine/Jilin/1-2/2011的HA和NA基因来源于野禽,与禽流感病毒的亲缘关系较近。其中,A/swine/Jilin/1-1/2011和A/swine/Jilin/1-2/2011的HA基因分别与韩国2004年分离到的A/wild duck/Korean/SH60/2004的H1N1亚型流感病毒和2005年中国河北分离到的A/duck/Hebei/843/2005的H1N2亚型流感病毒亲缘关系最近。同时,这两个病毒株的NA基因都与2003年中国东部的分离到的A/duck/Eastern China/103/2003和A/duck/Eastern China/152/2003的H1N1亚型流感病毒亲缘关系最近。与以上两个病毒株明显不同的是,A/swine/Jilin/1-3/2011的HA和NA基因则主要来自于人流感病毒和猪流感病毒,其中HA基因与中国山东、上海、广东、广西等地发现的H1N1病毒亚型亲缘关系最近,NA基因与中国山东、上海、广东等地发现的H1N1病毒亚型有明显的亲缘关系。
[Abstract]:In this study, 7909 pig serum samples, 5154 nasal swabs and suspected dead pigs were collected in Jilin area. A total of 13063 samples were isolated, and the subtypes of the isolated strains were identified. The results showed that the three swine flu viruses isolated were all H1N1 subtype swine flu viruses. According to the influenza virus naming standard system, they were named as A / R / J / J / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 / 1 and 1 / 3 / 1 / 1 / 1 respectively. Cloning and genetic evolution analysis of Hana gene were carried out. The result of gene sequencing showed that the HA gene sequence of Ar / Jilin / 1 / 1 / 2011H1 H1 was 1638 nucleotides, coding region was 40-1636bp. the coding length of 532 amino acids. Na gene was 1420 nucleotides, coding region was 4-1413bp, encoding 470 amino acids. The HA gene sequence of A / R / Jilin / 1 / 2 / 2011H1 / N1 was 1703 nucleotides, encoding 26-1556 BP, encoding 510 amino acids, encoding 1404 nucleotides, encoding 30-1397 BP, encoding 456 amino acids, and the HA gene sequence of A / R Jilinr / 1 / 3 / 2011H1 N _ 1 was 1745 nucleotides. The coding region is 17-1722 BP, encoding 568 amino acids. Na gene is 1458 nucleotides, encoding 21-1430 BP, encoding 470 amino acids. The cleavage sites showed that all of the three strains were low pathogenic, and the glycosylation sites were not mutated. The receptor binding sites showed that the isolates had the potential to infect avian origin. The resistance sites of na gene showed that the three swine influenza viruses were effective against the neuraminidase inhibitors currently used. Genetic evolution analysis showed that HA and na genes of A/swine/Jilin/1-1/2011 and A/swine/Jilin/1-2/2011 originated from wild birds and were closely related to avian influenza virus. Among them, A / R / J / J / 1 / 2011 and A/swine/Jilin/1-2/2011 HA genes were most closely related to A/wild duck/Korean/SH60/2004 's H1N1 subtype influenza virus isolated from Korea in 2004 and A/duck/Hebei/843/2005 's H1N2 subtype virus isolated from Hebei Province in 2005, respectively. At the same time, the na genes of the two strains were closely related to the A/duck/Eastern China/103/2003 and A/duck/Eastern China/152/2003 H1N1 subtype influenza virus isolated in eastern China in 2003. What is obviously different from the above two strains is that the HA and na genes of Ar / swine / Jilin / 1 / 3 / 2011 are mainly from human influenza viruses and swine flu viruses. Among them, HA genes are similar to those of Shandong, Shanghai, Guangdong, China. The relationship between the subtype of H1N1 virus found in Guangxi and the subtype of H1N1 virus found in Shandong, Shanghai and Guangdong provinces of China is very close to that of na gene.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.65

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