CAJ | 학술논문

目的了解上海地区2010年冬季人群甲型H1N1流行性感冒(流感)病毒流行株基因及抗原的变异.方法采集2010年12月至2011年1月间上海地区哨点医院流感样患者咽拭子标本137份,接种犬肾细胞(MDCK),分离流感病毒,直接免疫荧光法(DIF)鉴定流感病毒型,RT-PCR鉴定甲型H1N1,对部分甲型H1N1流感病毒株进行血凝素(HA)、神经氨酸酶(NA)、病毒聚合酶(PB2)片段全基因测序,分析基因及氨基酸位点变异.结果共分离到53株人流感病毒,48株为甲型H1N1流感病毒,按简单随机抽样法抽取19株测序.HA进化树分析发现,与2010年6月前分离的甲型H1N1毒株比较,绝大部分不位于同一主干上；HA蛋白的氨基酸位点分析显示,部分毒株在抗原决定位点上发生变异.NA蛋白酶活性中心及周围相关位点氨基酸组成保守,未检测到耐奥司他韦和扎那米韦的变异位点.PB2蛋白第627位和701位点分别是谷氨酸和天冬氨酸,仍是禽源流感病毒特征,但第677位点出现E677G突变.结论 2010年冬季上海地区人群甲型H1N1流感病毒流行株与之前春夏季分离株比较已经有一定变异,出现了一些抗原漂移和在哺乳动物宿主内的适应性进化.
목적 료해상해지구2010년동계인군갑형H1N1류행성감모(류감)병독류행주기인급항원적변이.방법 채집2010년12월지2011년1월간상해지구초점의원류감양환자인식자표본137빈,접충견신세포(MDCK),분리류감병독,직접면역형광법(DIF)감정류감병독형,RT-PCR감정갑형H1N1,대부분갑형H1N1류감병독주진행혈응소(HA)、신경안산매(NA)、병독취합매(PB2)편단전기인측서,분석기인급안기산위점변이.결과 공분리도53주인류감병독,48주위갑형H1N1류감병독,안간단수궤추양법추취19주측서.HA진화수분석발현,여2010년6월전분리적갑형H1N1독주비교,절대부분불위우동일주간상；HA단백적안기산위점분석현시,부분독주재항원결정위점상발생변이.NA단백매활성중심급주위상관위점안기산조성보수,미검측도내오사타위화찰나미위적변이위점.PB2단백제627위화701위점분별시곡안산화천동안산,잉시금원류감병독특정,단제677위점출현E677G돌변.결론 2010년동계상해지구인군갑형H1N1류감병독류행주여지전춘하계분리주비교이경유일정변이,출현료일사항원표이화재포유동물숙주내적괄응성진화.
Objective To understand the genetic and antigenic variations of influenza A (H1N1) isolates in Shanghai area in winter of 2010.Methods A total of 137 throat swabs were collected from patients with influenza-like illness in the sentinel hospital in Shanghai area from December 2010 to January 2011,then inoculated into Madin-Darby canine kidney (MDCK) cells.The types of influenza were identified by direct immunofluorescence assay (DIF) and influenza A (H1N1) subtype was determined by reverse transcriptase-polymerase chain reaction (RT-PCR).The mutations of gene and amino acid locus were analyzed through the whole genome sequencing of hemagglutinin (HA),neuraminidase (NA) and polymerase (PB2) segments from some influenza A (H1N1) isolates.Results Total of 53 human influenza virus strains were isolated including 48 influenza A (H1N1) virus strains.Nineteen strains were selected for sequencing by simple random sampling.The phylogenetic tree of HA gene revealed that the latest isolates and most of influenza A (H1N1) viruses isolated before June 2010 were not in the same stem.Analysis of amino acid residues in HA protein showed that mutations were found in antigenic determinant region in some strains.Residues at the enzyme active sites of NA protein were strictly conservative,no change was observed in amino acid residues which were related to drug resistance against oseltamivir and zanamivir.The 627 and 701 residues in PB2 protein were glutamic acid and aspartic acid,respectively,which was still the feature of avian influenza virus,but E677G mutation was detected.Conclusion Compared to influenza A (H1N1) strains isolated in spring and summer,some variations have been detected in the strains isolated in Shanghai area in winter of 2010,some antigen drift and adaptive evolution in mammalian hosts have appeared.