CAJ | 학술논문

目的探讨乙型流行性感冒病毒(简称乙型流感病毒)分离株血凝素(hemagglutinin,HA)、核蛋白(nucleoprotein,NP)、基质蛋白(matrix protein,M)和非结构蛋白(nonstructural protein,NS)基因的分子变异特征.方法选取浙江省1999-2012年乙型流感病毒分离代表株31株,进行HA重链区(HA1)、NP、M和NS基因测序,并构建基因进化树,估算此4个基因的核苷酸进化速率并分析其氨基酸变异位点.结果 31株乙型流感病毒分离株在HA1基因进化树上分别处于Victoria系和Yamagata系两大分支,分别以B/Victoria/2/87和B/Yamagata/16/88为代表.2010年之后Victoria系分离株的NP基因与Yamagata系毒株高度同源,处于同一进化分支.估算HA1 、NP、M和NS基因的核苷酸年进化速率分别为2.29×10-3、1.39×10-3、1.78×10-3、1.30×10-3/位点.Victoria系分离株HA1氨基酸重要变异位点主要包括K48E、L58P 、N75K 、K80R、K129N/S、N165K、S172P、S197N/D和A202V,Yamagata系分离株HA1的变异位点包括R48K、S150I、N166Y、N203S、G230D和D233N.2010年之后的Victoria系分离株NP氨基酸序列与Yamagata系毒株类似,与之前的毒株存在4个特征性氨基酸位点的变异,分别为A60D、L233V、N513S和V534I.结论 1999-2012年浙江省乙型流感病毒分离株发生明显变异,表面抗原基因HA1进化速度快于内部基因,基因重配和基因突变是病毒发生变异的主要机制.
목적 탐토을형류행성감모병독(간칭을형류감병독)분리주혈응소(hemagglutinin,HA)、핵단백(nucleoprotein,NP)、기질단백(matrix protein,M)화비결구단백(nonstructural protein,NS)기인적분자변이특정.방법 선취절강성1999-2012년을형류감병독분리대표주31주,진행HA중련구(HA1)、NP、M화NS기인측서,병구건기인진화수,고산차4개기인적핵감산진화속솔병분석기안기산변이위점.결과 31주을형류감병독분리주재HA1기인진화수상분별처우Victoria계화Yamagata계량대분지,분별이B/Victoria/2/87화B/Yamagata/16/88위대표.2010년지후Victoria계분리주적NP기인여Yamagata계독주고도동원,처우동일진화분지.고산HA1 、NP、M화NS기인적핵감산년진화속솔분별위2.29×10-3、1.39×10-3、1.78×10-3、1.30×10-3/위점.Victoria계분리주HA1안기산중요변이위점주요포괄K48E、L58P 、N75K 、K80R、K129N/S、N165K、S172P、S197N/D화A202V,Yamagata계분리주HA1적변이위점포괄R48K、S150I、N166Y、N203S、G230D화D233N.2010년지후적Victoria계분리주NP안기산서렬여Yamagata계독주유사,여지전적독주존재4개특정성안기산위점적변이,분별위A60D、L233V、N513S화V534I.결론 1999-2012년절강성을형류감병독분리주발생명현변이,표면항원기인HA1진화속도쾌우내부기인,기인중배화기인돌변시병독발생변이적주요궤제.
Objective To explore the characteristics of the genetic variation of hemagglutinin (HA) and three internal genes coding for the nucleoprotein (NP),matrix protein (M) and nonstructural protein (NS) of influenza B virus.Methods A total of 31 strains of influenza B virus were isolated in Zhejiang province from 1999 to 2012,and then were amplified and sequenced the genes of HA1,NP,M and NS.The phylogenetic tree was constructed,the nucleotide substitution rate of the above individual gene was estimated and the variation sites of amino acids were analyzed.Results The 31 isolated strains of influenza B virus were divided into two distinct lineages Victoria and Yamagata in the phylogenetic tree of HA1 gene,represented by B/Victoria/2/87 and B/Yamagata/16/88.Phylogenetic analysis of the NP gene showed that the NP gene of Victoria-like influenza B strains which were isolated after 2010 was highly homologous with Yamagata-like isolates,and thereby they were found to be on the same branch of the phylogenetic tree of the NP gene.Nucleotide substitution rates of HA1,NP,M and NS genes were estimated to be 2.29 × 10-3,1.39 × 10-3,1.78 × 10-3,1.30 × 10-3/site per year,respectively.Variations of amino acid of HA1 domain of Victoria-like isolates mainly included K48E,L58P,N75 K,K80R,K129N/S,N165 K,S172P,S197N/D and A202V; while those in Yamagata-like isolates were R48K,S1501,N166Y,N203S,G230D and D233N.Determined amino acid sequences of NP of Victoria-like influenza B isolates were similar to Yamagata-like isolates after 2010 and variations happened on four characteristic amino acid sites,naming A60D,L233V,N513S and V5341,compared with previous Victoria-like influenza B isolates.Conclusion Significant variation was found among influenza B strains isolated in Zhejiang province from 1999 to 2012.The surface HA1 gene evolved more rapidly than internal genes.Gene reassortment and gene mutation were the main evolutionary mechanism of influenza B virus.