[1]李建辉,刘春国,张云,等.甲型H1N1流感病毒株的分离鉴定以及PR8株的相关基因对重组病毒增殖能力的影响[J].中国预防兽医学报,2011,(07):517-520.
 LI Jian-hui,LIU Chun-guo,ZHANG Yun,et al.Isolation of an HIN1 influenza A virus and identification of genes influencing on the replication of reassortant virus with PR8 backbone[J].Chinese journal of preventive veterinary medicine,2011,(07):517-520.
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甲型H1N1流感病毒株的分离鉴定以及PR8株的相关基因对重组病毒增殖能力的影响
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年07期
页码:
517-520
栏目:
病原生物学
出版日期:
2011-07-15

文章信息/Info

Title:
Isolation of an HIN1 influenza A virus and identification of genes influencing on the replication of reassortant virus with PR8 backbone
作者:
李建辉;刘春国;张云;刘明;
中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部动物流感重点开放实验室;
Author(s):
LI Jian-hui LIU Chun-guo ZHANG Yun LIU Ming
State Key Laboratory of Veterinary Biotechnology, Animal Influenza Laboratory of the Ministry of Agricultural, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China
关键词:
甲型H1N1流感病毒反向遗传重配增殖能力
Keywords:
H1N1 influenza A virus reverse genetics reassortment replication capacity
摘要:
为分离流感病毒,本研究将疑似甲型流感患者的咽喉拭子接种SPF鸡胚尿囊腔,经检测收获的鸡胚尿囊液具有血凝活性。采用流感病毒通用引物进行PCR扩增出8个基因节段,经测序与GenBank中登录的序列比对后确定该病毒株为2009年爆发的甲型H1N1流感病毒,并命名为A/Harbin/LM/2009(H1N1),简称LM株。由于LM分离株在鸡胚中增殖能力比较弱,为提高其在鸡胚中的增殖能力,我们利用反向遗传技术将LM株的HA和NA基因与高增殖特性的人流感病毒细胞高产株PR8的另外6个节段进行"6+2"重配,获得拯救病毒,但该重配病毒血凝价并未显著提高。为进一步鉴定影响病毒增殖能力的基因,本研究将LM株所有节段以"7+1"的方式逐一与PR8的7个片段搭配构成完整的流感病毒基因组,分别得到8个重配病毒。结果显示分离株的PB1、PA、HA基因显著降低了重配病毒在鸡胚细胞中的增殖能力。
Abstract:
In this study, a virus was isolated from an adult throat swab in Harbin and identified as an H1N1 influenza virus by hemagglutination test and gene sequencing, designated A/Harbin/LM/2009 (H1N1). To improve the replication capacity of the isolate in embryoed eggs, we utilized reverse genetics technique to construct "6+2" reassortant virus, which included HA and NA segments of the isolate and the remaining segments of high-yield influenza virus PR8, as a result the HA titer of the reassortment virus was not significantly improved. To further identify which segment was critical to replication capacity of reassortant virus. A total of 8 reassortment viruses were constructed with 7 segments from PR8 and one segment from the isolate as a "7+1" strategy. The results showed that PB1, PA and HA segments of the isolate dramatically reduced the replication capacity of reassotant virus with PR8 backbone

参考文献/References:

[1] 郑薇,潘蔚绮,杨化强,李婷,张北武,李锋,陈凌. A/PR8/34(H1N1)NA基因包装信号对H5N1流感疫苗株NIBRG-14产量的影响[J]. 中国科学技术大学学报. 2010年04期
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[3] 刘明,张云,刘春国,潘蔚绮,刘超男,杨涛. 利用反向遗传学技术构建H5亚型禽流感高产疫苗株[J]. 生物工程学报. 2006年05期
[4] Webby R J,Perez D R,Coleman J S,et al. Responsiveness to a pandemic alert: use of reverse genetics for rapid development of influenza vaccines. The Lancet. 2004
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[6] Liu M,Wood J M,Ellis T. Preparation of a standardized efficacious agricultural H5N3vaccine by reverse genetics[J]. Journal of Virology. 2003

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备注/Memo

备注/Memo:
基金:黑龙江省自然科学基金(ZJN0702-02);;现代农业水禽技术体系(CARS-43)
更新日期/Last Update: 2011-09-29