乙型肝炎病毒前S1抗原关键表位基因分型及其与全长前S1抗原基因型的相关性  

HBV genotyping based on key epitopes of PreS1 antigen and its correlation with genotyping by fulllength PreS1 sequencing

作　　者：张晓晨 李玉敏 李家亿 康新迪 何欣悦 牛俊奇[2] 温晓玉[2] 刘镇宁 Zhang Xiaochen;Li Yumin;Li Jiayi;Kang Xindi;He Xinyue;Niu Junqi;Wen Xiaoyu;Liu Zhenning(Key Laboratory of Bionic Engineering （Ministry of Education）, Jilin University, Changchun 130000, China;the First Hospital of Jilin University, Changchun 130000, Chin)

机构地区：[1]吉林大学工程仿生教育部重点实验室,长春130000 [2]吉林大学白求恩第一医院,130000

出　　处：《中华肝脏病杂志》2018年第5期371-376,共6页Chinese Journal of Hepatology

基　　金：艾滋病和病毒性肝炎等重大传染病防治科技重大专项（SQ2017ZX106080）;吉林省科技发展计划国际科技合作项目（20150414016GH）;吉林省医药产业发展专项资金项目（20130727033YY）

摘　　要：目的 明确乙型肝炎病毒（HBV）大表面蛋白前S1区（PreS1）两个重要表位在中国乙型肝炎患者群中的基因型分布,探讨两表位基因型之间及两表位基因型与PreS1全长基因型之间的相关性,确定能否根据关键表位的多肽序列判断PreS1全长基因型. 方法 从患者血清中提取HBV DNA进行PCR扩增,对扩增成功的278例样本进行DNA测序,并与GenBank中已知的HBV序列比对,确定HBV大表面蛋白PreS1的两个关键表位（第21 ～ 47号氨基酸表位和第94 ～ 117号氨基酸表位,分别简称为P21表位和P94表位）基因型及PreS1全长基因型,并对3组基因分型结果进行一致性分析.一致性检验采用Kappa分析. 结果 成功测序232例样本.根据P21表位蛋白序列的分型结果为：C基因型201例,B基因型23例,基因型不确定8例.根据P94表位蛋白序列的分型结果为：C基因型199例,B基因型25例,基因型不确定8例.根据PreS1全长序列的分型结果为：C基因型207例,B基因型25例.P21表位基因分型和P94表位基因分型与PreS1全长基因分型结果均高度一致,分别为96.55％和96.12％（Kappa值分别为0.841,0.826）,且两表位的基因分型结果也具有很好的一致性（93.10％,Kappa值为0.718）. 结论 本研究创新性地提出基于关键表位氨基酸序列的基因分型方法,验证了中国乙型肝炎患者群的HBV基因型以B和C基因型为主,且HBV PreS1关键保守表位的基因分型结果与全长基因分型结果具有高度一致性（＞96％）,可以采用一个或两个关键保守表位的基因分型代替全长基因分型.Objective The aim was to investigate the genotype distribution of two major epitopes of large surface protein （PreS1） of hepatitis B in Chinese patients and to explore the association between the genotypes of these two epitopes,and to determine whether PreS1 full-length genotype could be revealed according to the polypeptide sequence of key epitopes.Methods HBV DNA was extracted from the serum of patients for PCR amplification.278 samples amplified successfully were sequenced and compared with the known HBV sequences in Genbank to determine the two key epitopes of HBV PreS1 genotype （amino acid epitope 21-47 and 94-117,abbreviated as P21 and P94） and PreS 1 full-length genotypes.The correlation among three genotyping approaches was analyzed by Cohen＇s kappa coefficient to verify the consistency between the key-epitope genotyping and the full-length preS1 genotyping.Results 232 samples were successfully sequenced.The genotyping based on the kind of P21 epitope protein sequence,201 cases for genotype C,23 cases for genotype B and 8 cases for uncertain genotypes and genotyping based on the form of P94 epitope protein sequence,199 cases for genotype C,25 cases for genotype B and 8 cases for indeterminate genotypes.Lastly,the genotyping based on sequence of the full-length PreS1 sequence,207 and 25 cases for genotype C and B.P21 or P94 epitope genotyping and PreS 1 full length genotyping were highly consistent,respectively,96.55% and 96.12%,and the two epitopes （P21 and P94） genotyping have parallel consistency （93.10%）.Conclusion In this study,an innovatively genotyping method based on the amino acid sequence of key epitopes was proposed.The genotypes of HBV in china were mainly B and C genotypes,and the genotypes of key conserved epitopes of HBV PreS 1 were highly consistent with the full-length genotyping （〉 96%）.Moreover,genotyping with one or two key epitopes can be used in place of the full-length genotyping.

关 键 词：肝炎病毒 乙型 基因分型 前S1区 表位 蛋白序列 

分 类 号：R512.62[医药卫生—内科学]