黄吉城1,钟玉清1,师永霞1,刘丽珊1,2,丁国允1,郑夔1,李小波1,苏锦坤1,幸芦琴1,洪烨1

　　1. 广东出入境检验检疫局检验检疫技术中心, 广东  广州  510700; 2.广州市黄埔区食品药品监督管理局

　　摘要:目的   分析口岸输入性甲型H1N1流感病毒神经氨酸酶(NA)基因特征及变异趋势, 及早发现变异株。方法  选取广东口岸入境人员中检出的64份输入性甲型H1N1流感阳性样本,对其病毒NA基因进行核苷酸序列测定。与2009年甲型H1N1流感代表株A/California/04/2009/H1N1的NA基因序列进行对比,对甲型H1N1流感病毒的NA基因进化树、核苷酸序列同源性、氨基酸序列的变异及蛋白分子结构变化等进行分析。结果  NA基因进化树分为3个分支, 其中2009年、2010年病毒聚集成簇位于分支1,2011年病毒则与部分2010年检出的病毒位于较远距离的分支2和3。检出的1例病毒NA氨基酸的抗原决定簇上发生Y155H的变异,部分病毒NA蛋白在42位点上新增了一个糖基化位点,在68、386位点出现了糖基化位点的缺失。氨基酸G41R、V106I、V241I、N248D、I365T、N369K等位点的变异较为显著,但未见耐奥司他韦病毒的出现。结论   甲型H1N1流感病毒NA基因已发生一定程度的变异。随着突变的不断积累,病毒极可能会通过改变其抗原性和耐药性, 从而引起新的流行。

　　关键词:甲型H1N1流感病毒; NA基因; 变异; 输入性; 分析

　　中图分类号:R511.7   文献标识码:B

　　Analysis on neuraminidase (NA )gene variation

　　in imported influenza A(H1N1) viruses

　　HUANG Ji-cheng*, ZHONG Yu-qing, SHI Yong-xia, LIU Li-shan, DINGGuo-yun,

　　ZHENG Kui, LI Xiao-bo, SU Jin-kun, XING Lu-qin, HONG Ye

　　*Guangdong inspection and quarantine technology center,Guangzhou, Guangdong 510700, China

　　Abstract:   Objective   To analyze the characteristicsand mutation tends of neuraminidase (NA) gene in imported influenzaA(H1N1) viruses, and to discover variant strain early.  Methods   A total of 64 positive samples of influenza A(H1N1)viruses were detected from the entry cases at Guangdong ports. Thenucleotide of NA gene were amplified by RT-PCR, then sequenced.Phylogenetic tree, nucleotide sequence homology, amino acidsequence homology and structural changes in proteins were analyzedby comparing with NA gene of A/California/04/2009/H1N1.  Results   Phylogenetic tree of NA genes were divided intothree lineages. Viruses detected in 2009 and 2010 were clusteredtogether to lineages I, while all of viruses in 2011 and part in2010 were located to the distant lineages II and III. NA aminoacids epitopes of one case occured on Y155H variation. NA proteinsof some viruses appeared to add a glycosylation sites at site 42and miss their glycosylation sites at sites 68 and 386. Meanwhile,there were more significant amino acids variation at sites G41R,V106I, V241I, N248D, I365T and N369K, but no Oseltamivir-resistantvirus was observed.   Conclusion   Variations of NA inpandemic influenza A(H1N1) viruses occurred from 2009 to 2011. Withthe continued accumulation of mutations, the virus will likely tochange its antigenic and cause a new epidemic.

　　Key words:   Influenza A(H1N1) virus; NA gene; Variation;Import; Analyze《中国国境卫生检疫杂志》2014年2期