CAJ | 학술논문

目的从北京地区一例急性期甲肝患者血清中提取甲肝病毒核酸,获得一株甲肝病毒(HAV)全基因组序列,进行相关分析.方法分8段设计16对重叠基因片段引物,经提取病毒RNA、反转录、巢式PCR扩增及产物直接测序获得序列.结果获得的BJ73全基因组核苷酸全长7477 bp,编码含2227个氨基酸的多聚蛋白;同源比对发现BJ73核苷酸和氨基酸序列均与LU38同源性最高,分别为99.1％和99.7％;VP3-VP1区域内已发表的抗原中和位点处未发现氨基酸变异;分别基于全基因序列和不同区段序列构建系统进化树,均表明B J73病毒株属于IA亚型;选择压力分析表明全编码区处于负向选择.结论本实验得到的相关数据为我国甲肝病毒基因型分布、病毒进化等研究提供了依据和实验基础.
목적 종북경지구일례급성기갑간환자혈청중제취갑간병독핵산,획득일주갑간병독(HAV)전기인조서렬,진행상관분석.방법 분8단설계16대중첩기인편단인물,경제취병독RNA、반전록、소식PCR확증급산물직접측서획득서렬.결과 획득적BJ73전기인조핵감산전장7477 bp,편마함2227개안기산적다취단백;동원비대발현BJ73핵감산화안기산서렬균여LU38동원성최고,분별위99.1％화99.7％;VP3-VP1구역내이발표적항원중화위점처미발현안기산변이;분별기우전기인서렬화불동구단서렬구건계통진화수,균표명B J73병독주속우IA아형;선택압력분석표명전편마구처우부향선택.결론 본실험득도적상관수거위아국갑간병독기인형분포、병독진화등연구제공료의거화실험기출.
Objective To identify the complete nucleotide sequence of hepatitis A virus (HAV) extracted from serum of an acute hepatitis A patient in 2007 (B J73) in Beijing.Methods The full genomic sequence was sequenced and subjected to the phylogenetic analysis using Bioedit 7.0,MEGA5 and HyPhy.Results The genome of BJ73 consisted of 7477 nucleotides,encoding a polyprotein of 2227 amino acids.Sequence comparison showed that B J73 shared the highest identities of 99.1％ for nucleotide and 99.7％ for amino acid with LU38.Amino acid change was not observed in reference to published neutralizing antigenic sites in VP3-VP1 regions.Phylogenetic analysis included the B J73 sequence in subgenotype IA,either considering different fragments or the full length nucleotide sequence; negative selection was found in coding regions.Conclusion This article provides new data on the distribution of genotype and virus evolution of HAV isolated from China.