H5N1亚型禽流感病毒A/Guangxi/1/2005株抗药机制的研究  

Mechanism of the H5N1 subtype influenza virus A/Guangxi/1/2005 resistance to antiviral drug

作　　者：王飞[1] 吴珊珊[1] 慈彦鹏[1] 刘丽玲[1] 田国彬[1] 曾显营[1] 管雪婷[1] 李雁冰[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/国家禽流感参考实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2012年第9期682-685,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：农业部行业专项-边境动物疫病防治技术研究(20110308);农业部动物流感监测与防制专项(20110308)

摘　　要：为研究H5N1亚型禽流感病毒(AIV)的抗药机制,本研究选取Clade2.3.4亚群中一株对金刚烷胺敏感的人源AIV A/Guangxi/1/2005(H5N1)(S-GX05),用抗流感病毒药物金刚烷胺对其进行定向诱导,筛选出一株抗药性病毒株,命名为R-A/Guangxi/1/2005(R-GX05)。通过全基因测序并与S-GX05全基因序列进行对比,结果显示S-GX05只在其M2蛋白中有一个氨基酸位点发生突变,即A30P;抗药性鉴定这两株病毒的半数药物抑制浓度(IC50)分别为0.9μM和48.9μM,表明R-GX05对金刚烷胺表现出一定程度的抗性。动物实验证实,这两株病毒对BALB/c小鼠的致病性基本一致,均表现出高致病性,其MLD50分别为4.7 log10 EID50和5.0 log10 EID50,两株病毒在小鼠体内各组织脏器中的分布及增殖能力也基本相同。这些结果表明,S-GX05在药物压力下产生抗药性后,并未引起其它生物学特性的改变。A30P的发现为进一步从分子水平上研究H5N1亚型AIV的抗药机制及新型抗流感新药的研发奠定了基础。To investigate the mechanism of influenza virus resistance to antiviral drug of Amantadine, the highly pathogenic H5N1 subtype avian influenza virus （AIV） A/Guangxi/1/2005 （S-GX05） was induced with 1,000 μM of Amantadine for 15 passages in MDCK cell culture and the resulting virus was converted from sensitive to resistant for Amantadine （designate R-GX05）. Sequencing analysis showed that only one point mutation occurred in M2 encoding sequence and resulted in a substitution of A^30p in M2 protein of R-GX05. The inhibition concentration （IC50） of Amantadine to S-GX05 and R-GX05 was 0.9 μM and 48.9 μM, respectively. However, the pathogenicity tests indicated that both viruses were highly pathogenic in mice with a median lethal dose （LD50） of 4.7 log10 EID50 and 5.0 log10 EID50, respectively. The tissue tropism and replication capacities of both viruses had no difference. In conclusion, Amantadine-resistant M2 mutants of AIV A/Guangxi/1/2005 had no change in growth characteristics or virulence in mice. The finding of A^30P mutation would pave the way to further research focusing on the molecular mechanisms of influenza virus resistant to antiviral drugs.

关 键 词：H5N1禽流感病毒 抗药性 金刚烷胺 

分 类 号：S852.659.5[农业科学—基础兽医学]