CAJ | 학술논문

目的分析两个流行性感冒(流感)流行季甲型H1N1流感病毒pdm09和H3N2流感病毒的核苷酸和氨基酸变异特点.方法在2012年12月至2013年2月和2013年12月至2014年2月两个流感流行季,对反转录PCR证实的54株流感病毒株进行分型和血凝素(HA)、神经氨酸酶(NA)扩增并测序,从GenBank获得2009年以来全球范围包括北京地区的甲型H1N1流感病毒pdm09和H3N2流感病毒序列,用Bioedit7.09和Mega5.05软件进行比对并构建系统进化树.结果共分离到甲型H1N1流感病毒pdm09 35株,H3N2流感病毒19株.甲型H1N1流感病毒pdm09株的HA和NA基因在进化树上的聚集情况基本一致,2012年至2013年的H3N2流行株位于3C亚群.北京地区2009年至2014年甲型H1N1流感病毒pdm09表面抗原HA和NA氨基酸差异分别为0.3％～15.2％和0.5％～3.3％;2012年至2013年的HA抗原表位发生L178I、S202T和H155Q/R位点突变,分别位于抗原表位Sa、Sb和Ca; 2013年至2014年又新增K180Q和G187E位点突变.2009年至2014年H3N2流感病毒表面抗原HA氨基酸差异为0～7.8％,与疫苗株A/Texas/50/2012相比,北京地区2012年至2014年H3N2流感病毒表面抗原HA氨基酸差异为0～0.7％.2012年至2013年H3N2流感病毒HA抗原表位突变位点为R158K/G、N161S和P214S,位于抗原表位A和B;2013年至2014年新增I208V和I64V突变位点.未发现奥司他韦耐药毒株.结论 2012年至2014年北京地区甲型流感病毒发生新的位点突变,历年流行的病毒株表面抗原(尤其是HA)差异范围较大.
목적 분석량개류행성감모(류감)류행계갑형H1N1류감병독pdm09화H3N2류감병독적핵감산화안기산변이특점.방법 재2012년12월지2013년2월화2013년12월지2014년2월량개류감류행계,대반전록PCR증실적54주류감병독주진행분형화혈응소(HA)、신경안산매(NA)확증병측서,종GenBank획득2009년이래전구범위포괄북경지구적갑형H1N1류감병독pdm09화H3N2류감병독서렬,용Bioedit7.09화Mega5.05연건진행비대병구건계통진화수.결과 공분리도갑형H1N1류감병독pdm09 35주,H3N2류감병독19주.갑형H1N1류감병독pdm09주적HA화NA기인재진화수상적취집정황기본일치,2012년지2013년적H3N2류행주위우3C아군.북경지구2009년지2014년갑형H1N1류감병독pdm09표면항원HA화NA안기산차이분별위0.3％～15.2％화0.5％～3.3％;2012년지2013년적HA항원표위발생L178I、S202T화H155Q/R위점돌변,분별위우항원표위Sa、Sb화Ca; 2013년지2014년우신증K180Q화G187E위점돌변.2009년지2014년H3N2류감병독표면항원HA안기산차이위0～7.8％,여역묘주A/Texas/50/2012상비,북경지구2012년지2014년H3N2류감병독표면항원HA안기산차이위0～0.7％.2012년지2013년H3N2류감병독HA항원표위돌변위점위R158K/G、N161S화P214S,위우항원표위A화B;2013년지2014년신증I208V화I64V돌변위점.미발현오사타위내약독주.결론 2012년지2014년북경지구갑형류감병독발생신적위점돌변,력년류행적병독주표면항원(우기시HA)차이범위교대.
Objective To analyze the variation characteristics of nucleotide and amino acid of influenza A viruses subtype H1N1 pdm09 and H3N2 during influenza season.Methods Fifty-four strains of influenza A virus strains were obtained and confirmed by reverse transcription polymerase chain reaction (PCR) during two influenza seasons including from Dec 2012 to Feb 2013 and from Dec 2013 to Feb 2014.Hemagglutinin (HA) and neuraminidase (NA) genes of these strains were amplified by PCR and then sequenced.The aligned sequences in other regions of China and other countries were accessed from GenBank.Bioedit 7.09 and Mega 5.01 software were used for alignment and phylogenetic tree analysis.Results A total of 35 strains of H1N1 pdm09 and 19 strains of H3N2 were isolated.The coloured HA clusters in the phylogenetic tree coincided with those of NA.The epidemic strains of H3N2 during 2012 and 2013 located in 3C subgroup.For A(H1N1)pdm09 between 2009 and 2014 in Beijing,the amino acid differences of surface antigens HA and NA were 0.3％-15.2％ and 0.5％-3.3％,respectively.Mutations HA-L178I,S202T,H155Q/R were located at antigenic sites Sa,Sb and Ca of A(H1N1) pdm09 during 2012 to 2013,while new mutations HA-K180Q and G187E were found during 2013 to 2014.For H3N2,the amino acid difference of antigen HA between 2009 and 2014 was 0-7.8％.Compared to vaccine strain A/Texas/50/2012,the amino acid difference of surface antigen HA during 2012 to 2014 in Beijing was 0-0.7％.Mutations HA-R158K/G,N161S and P214S were located at antigenic sites A and B of H3N2 during 2012 to 2013,while new substitutions HA-I208V and 164V were observed during 2013 to 2014.All of the strains were sensitive to Oseltamivir.Conclusions New mutations are found in the HA and NA genes of influenza A virus during influenza season between 2012 and 2014 in Beijing.There is great difference in surface antigens among the epidemic influenza A strains over the years,especially for HA gene.