H5N1亚型人禽流感病毒A/Anhui/2/2005株反向遗传操作系统的建立  

Establishment of reverse genetics system of the human H5N1 influenza virus isolate A/Anhui/2/2005

作　　者：冯华朋[1] 钟功勋[1] 张月梅[1,2] 李旭勇[1] 李雁冰[1] 施建忠[1] 陈化兰[1] 

机构地区：[1]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,黑龙江哈尔滨150001 [2]内蒙古农业大学动物科学与医学学院,内蒙古呼和浩特010018

出　　处：《中国预防兽医学报》2010年第2期135-138,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家973项目(2005CB523005);国家杰出青年基金(30825032)

摘　　要：为建立H5N1亚型人源禽流感病毒A/Anhui/2/2005(W-AH05)反向遗传操作系统,本研究构建了W-AH05的8重组质粒,拯救出人源禽流感病毒R-A/Anhui/2/2005(R-AH05)。全基因序列测定结果表明,救获病毒R-AH05与野生病毒W-AH05的核苷酸序列完全一致;动物试验证实,R-AH05保持了W-AH05的对BALB/c小鼠呈高致病力的特性,其MLD50分别为1.5log10EID50和1.2log10EID50。R-AH0与W-AH05分别以106EID50剂量鼻腔感染BALB/c小鼠,病毒在小鼠体内的分布情况及各个脏器中的病毒滴度基本相同。由此可见,R-AH05保持了W-AH05的生物学特性,从而为进一步研究人源禽流感病毒的致病机理及跨宿主传播机制等提供了操作平台。To obtain the rescued virus of the H5N1 avian influenza virus from human isolate A/Anhui/2/2005 （W-AH05）, eight plasmid system was established, and the rescued virus R-AH05 was generated by reverse genetic technique. The full genome of the R-AH05 had no nucleic acid change, compared with W-AH05 through sequence analysis. R-AH05 had the same pathogenicity as the parent virus in mice with 50 % mouse lethal dose （MLD50） of 1.5 log10EID50 and 1.2 log10EID50, respectively. The tissue tropism and virus load of R-AH05 and W-AH05 were highly identical in respective tissues of the mice infected intranasally with 10^6 EID50 dose. In conclusion, the rescued virus R-AH05 had congruously biological characteristics with the wild type virus W-AH05. The results could provide a technique for further research on the pathogenic and crossing-host-barrier mechanisms of the HSN 1 avian influenza virus from human isolate.

关 键 词：H5N1 人源禽流感病毒 反向遗传操作 

分 类 号：S852.65[农业科学—基础兽医学]