CAJ | 학술논문

目的了解2001-2006年间辽宁省甲3亚型流感病毒HAl基因变异特征.方法采用辽宁省流感监测的咽拭分离株,对其中的7株甲3病毒核酸提取,经逆转录-聚合酶链反应(RT-PCR),扩增产物测序,推导出其氨基酸序列,并用DNA MAN软件包中的Alignment做了差异和同源性比较,用Mega软件包中的Neighbor Joining方法 ,通过与NCBI数据库中2000-2006年28株H3N2流感病毒同源比对后,制作种系进化树,进行基因进化特征分析.结果序列分析发现,甲3亚型流感病毒与WHO当年的疫苗株M/California/7/2004比较,在其HA1核酸序列上有12处碱基不同,4个氨基酸位点发生了替换.基因及氨基酸序列同源性下降.与WHO北半球疫苗株A/Wisconsirn/67/2005序列更接近,但仍有4处碱基不同.结论甲3亚型流感病毒基因序列发生了变异,与甲3亚型疫苗株A/California/7/2004同源性有下降趋势,提示其抗原可能发生部分漂移或更换,与南方浙江2005年株、WHO北半球2006-2007疫苗株A/Wiseonsin/6712005序列接近,提示南方春夏季流感流行株极有可能在北方秋冬季流行,对流感流行病学监测,疫苗株使用指导都具有重要意义.
목적 료해2001-2006년간요녕성갑3아형류감병독HAl기인변이특정.방법 채용요녕성류감감측적인식분리주,대기중적7주갑3병독핵산제취,경역전록-취합매련반응(RT-PCR),확증산물측서,추도출기안기산서렬,병용DNA MAN연건포중적Alignment주료차이화동원성비교,용Mega연건포중적Neighbor Joining방법 ,통과여NCBI수거고중2000-2006년28주H3N2류감병독동원비대후,제작충계진화수,진행기인진화특정분석.결과 서렬분석발현,갑3아형류감병독여WHO당년적역묘주M/California/7/2004비교,재기HA1핵산서렬상유12처감기불동,4개안기산위점발생료체환.기인급안기산서렬동원성하강.여WHO북반구역묘주A/Wisconsirn/67/2005서렬경접근,단잉유4처감기불동.결론 갑3아형류감병독기인서렬발생료변이,여갑3아형역묘주A/California/7/2004동원성유하강추세,제시기항원가능발생부분표이혹경환,여남방절강2005년주、WHO북반구2006-2007역묘주A/Wiseonsin/6712005서렬접근,제시남방춘하계류감류행주겁유가능재북방추동계류행,대류감류행병학감측,역묘주사용지도도구유중요의의.
Objective To understand the HA1 genetic variation characterization of influenza virus subtype H3N2 circulated from 2001 to 2006 in Liaoning local area. Methods Viral RNA was extracted and transcribed into cDNA by reverse transcriptase and amplified by PCR. The product of PCR was purified by QIAgen purification kits,and sequenced by ABI 3100avant. The sequence data were analyzed phylngeneticaUy by Sequence software with epidemic records. Finally, the phylngenetic trees were drawn according to deduced amino acid sequences of influenza vires H3N2 from 2000 to 2006 in the NCBI database. Results The seven HAldomain sequences of H3N2 influenza viruses circulated from 2001 to 2006 in Liaoning local area had been analyzed. Compared with WHO 2004-2006 H3N2 vaccine A/California/7/2004, 12 bases had changed, 4 positions had amino acid substitution in 62 *>E, 182 T>1,224 S>A ,225 C>Y. 224 and 225 are RBS (Receptor binding site). The homology is lower than 98%. Phylogenetic tree showed Liaoning H3N2 2006 strains and Zhejiang 2005 strains were similar to WHO Northern hemisphere winter 2006-2007 Vaccine A/Wisconsin/67/2005 (H3N2)-like virus and grouped together to form an independent cluster even though several bases were still different. Conclusion The HA1 domain of HA gene of influenza viruses (H3N2) isolated from 2001-2006 in Liaoning local area showed base mutation, amino acid sequence difference compared to A/California/7/2004 (2005-2006 vaccine), suggesting it might be the main cause leading to the spread of influenza. The sequence analysis showed Liaoning 2006 H3N2 strains were similar to those from Southern area which suggested that furher surveullance should be conducted to monitor the virus mutation in circulation.