青海省2010～2012年H3N2型流感病毒NA基因特性研究  

Genetic Characteristics of Influenza A/H3N2 Virus Neuraminidase Gene:A Survey from 2010 to 2012 in Qinghai Province,China

作　　者：于娟[1] 饶华祥[1] 卢囡囡 李红[1] 易虎[1] 赵生仓 

机构地区：[1]青海省疾病预防控制中心,西宁810007

出　　处：《病毒学报》2014年第3期263-267,共5页Chinese Journal of Virology

基　　金：国家"十一五"科技重大专项(2009ZX10004-208)

摘　　要：为了解2010～2012年青海省H3N2亚型流感病毒神经氨酸酶（NA）基因变异特征，并探讨对NA抑制剂的耐药性情况，本研究随机选择2010～2012年流感病原监测中分离到的H3N2亚型流感毒株，提取病毒RNA，通过RT-PCR法扩增病毒NA基因，纯化产物进行核苷酸序列测定，采用MEGA4.0软件对其核苷酸序列及所推导的氨基酸序列进行基因特性分析。绘制NA基因进化树，研究发现2010～2011年H3N2型分离株与2010～2012年世界卫生组织（World Health Organization， WHO）推荐疫苗株A/Perth/16/2009和2008～2010年WHO推荐疫苗株A/Brisbane/10/2007聚集成簇，处于同一进化分支，2012年分离株则独立形成另一进化分支。将核苷酸序列推导成氨基酸序列，与2010～2012年WHO推荐疫苗株A/Perth/16/2009相比，2010年H3N2亚型分离株氨基酸变异位点在K81T；2011年H3N2亚型分离株氨基酸变异位点在I26V和D127N；2012年H3N2亚型分离株氨基酸位点变异在E41K，P46A，I58V，T71N，L81P，D93G，D127N，D151N和I307M，第151位处于NA蛋白酶活性中心，D151N变异使分离株增加了一个糖基化位点。上述结果表明青海省2010~2011年H3N2亚型流感病毒NA基因未发生明显变异，2012年H3N2亚型流感病毒则发生了较大变异，可能会对NA抑制剂扎纳米韦和奥司他韦产生轻微耐药性。This study aims to perform a survey of genetic variation in neuraminidase （NA） gene of influenza A/H3N2 virus, as well as related resistance to NA inhibitors, in Qinghai Province of China, 2010 to 2012. Strains of influenza A/H3N2 isolated during an influenza survey from 2010 to 2012 in Qinghai were enrolled by random sampling. Viral RNA was extracted and amplified by RT-PCR. Purified PCR products were sequenced thereafter. Genetic analysis of nucleic acid and the derived amino acid sequences was performed by MEGA 4.0. Phylogenetic trees were also constructed. Strains isolated during 2010-2011 in this study clustered closely with World Health Organization （WHO） 2010-2012 reference vaccine strain A/ Perth/16/2009 and 2008-2010 reference vaccine strain A/Brisbane/10/2007 on the phylogenetic tree, while the 2012 isolates were located on another branch. In analysis of derived amino acid sequences, the 2010 isolates mutated at K81T, the 2011 isolates mutated at I26V and D127N, while the 2012 isolates mutated at E41K, P46A, I58V, T71N, L81P, D93G, D127N, D151N, and I307M. The D151N mutation added a glycosylation site to the activity center of NA. No significant variation was discovered in H3N2 NA gene of 2010-2011 isolates in Qinghai, China. Isolates of 2012 were found with significant mutation, which has the potential of inducing minor resistance to NA inhibitors like zanamivir and oseltamivir.

关 键 词：流感病毒 H3N2亚型 神经氨酸酶 耐药性 

分 类 号：R373.9[医药卫生—病原生物学]