表面抗原和表面抗体检测双阳性CD重组型乙型肝炎病毒全基因组序列分析  

Whole-Sequence Analyses of HBV/CD Recombinants in Patients with Coexisting HBsAg and HBsAb

作　　者：刘贺 沈立萍[1] 张爽[1] 王锋[1] 张国民[2] 梁晓峰[3] 王富珍[2] 毕胜利[1] LIU He;SHEN Liping;ZHANG Shuang;WANG Feng;ZHANG Guomin;LIANG Xiaofeng;WANG Fuzhen;BI Shengli(NHC Key Laboratory for Medical Virology,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;National Immunization Program,Chinese Center for Disease Control and Prevention,Beijing 100052,China;Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,国家卫生健康委员会医学病毒和病毒病重点实验室,北京102206 [2]中国疾病预防控制中心免疫规划中心,北京100052 [3]中国疾病预防控制中心,北京102206

出　　处：《病毒学报》2020年第3期400-406,共7页Chinese Journal of Virology

基　　金：国家科技重大专项(项目号:2017ZX10105015001002),题目:慢性乙型病毒性肝炎血清和分子流行特征研究。

摘　　要：西藏地区藏族人群乙型肝炎病毒(Hepatitis B virus,HBV)感染率较高,而针对感染者血清中HBV表面抗原(Hepatitis B surface antigen,HBsAg)和HBV表面抗原抗体(Hepatitis B surface antibody,HBsAb)双阳性的研究一直进展缓慢,尚无明确的研究结论。为探讨西藏地区藏族人群慢性HBV感染者血清中HBsAg和HBsAb双阳性与基因组核苷酸/氨基酸突变的关系,本研究在西藏选取7个地区作为研究区域,进行多阶段抽样,选取样本进行HBV血清五项指标检测,筛选HBsAg和HBsAb均为阳性的患者血清共24份作为双阳性组,以年龄和乙型肝炎e抗原(HBeAg)等感染指标进行匹配,选取96份HBsAg阳性,HBsAb阴性患者血清作为对照组。HBV全基因组序列通过聚合酶链式反应(Polymerase chain reaction,PCR)产物直接测序获得,并进行重组分析和突变分析。852名西藏HBV感染者中,HBsAg/HBsAb双阳性率为2.82%(24/852)。双阳性组在S蛋白N端和主要亲水区(Major hydrophilic region,MHR)的突变率以及PreS缺失发生率均显著高于对照组。T1753C、C1990T和C2002T等核苷酸突变;S蛋白中V224A、PreS区D103E等氨基酸突变在两组内分布存在显著差异。HBV/CD重组型的HBsAg/HBsAb双阳性发生率与中国乙肝主要流行区域接近。HBV感染者血清HBsAg和HBsAb共存可能与S蛋白,特别是MHR内的高氨基酸突变造成的免疫逃逸有关。PreS缺失、S抗原蛋白C端V224A突变和PreS区D103E突变可能对HBsAg/HBsAb双阳性的产生具有协同作用。Hepatitis B virus(HBV)infection is at a high level in the Tibetan population,while studies on the double positivity of HBV surface antigen(HBsAg) and HBV surface antibody(HBsAb) in the serum of infected persons were in slow progress and without a conclusion. To study the relationship between the HBsAg and HBsAb coexistence in Tibetan people with chronic HBV infection and the HBV nucleotide/amino acid mutations. Seven regions in Tibet,China,were selected as the study area for multi-stage sampling. Samples were selected for detection of five HBV serum markers. Twenty-four serum samples with HBsAg and HBsAb coexistence were selected as the double-positive group,while ninety-six HBsAg positive and HBsAb negative serums which comparable with double-positive group in age and Hepatitis B e antigen(HBeAg)status were selected as the control group. Whole-genome sequences of HBV was obtained by direct sequencing of Polymerase Chain Reaction(PCR)products,which were under recombination analysis and mutation analysis.The frequency of HBsAg/HBsAb coexistence was 2.82%(24/852). The mutation rate of N-terminal,major hydrophilic region(MHR)and the incidence of PreS deletion in the double-positive group were significantly higher than those in the control group. The nucleotide mutations of T1753C,C1990T and C2002T,and the amino acid mutations of V224A in S protein and D103E in PreS region were significantly different between the two groups. The frequency of HBsAg/HBsAb double positive in HBV/CD recombinant is close to the main genotype prevalence in other parts of China. The coexistence of serum HBsAg and HBsAb in HBV infected patients may be related to the immune escape caused by high amino acid variation in S protein,especially MHR region. The PreS deletion,V224A mutation in the C-terminal of S protein and D103E mutation in the PreS region may have synergistic effect on the occurrence of HBsAg/HBsAb coexistence.

关 键 词：乙型肝炎病毒(HBV) 乙型肝炎病毒表面抗原(HBsAg) 抗乙型肝炎病毒表面抗原抗体(HBsAb) 突变 

分 类 号：R373.2[医药卫生—病原生物学]