慢性乙型肝炎肝肾阴虚证HBV基因突变点的分布规律

目的:研究慢性乙型肝炎肝肾阴虚证HBV基因突变点的分布规律。方法:基因芯片方法检测慢性乙型肝炎肝肾阴虚证患者HBV前C区基因变异。结果:慢性乙型肝炎肝肾阴虚证患者HBV前C区变异可见于多个位点,1896位点变异率为31.37%,1764位点变异率为25.49%,1762位点变异率为24.51%,1899位点变异率为12.75%,1862位点变异率为5.88%。1762、1764、1896和1899点的突变株信号值高于野生株信号值;1862点野生株信号值高于突变株信号值,差异有显著性意义(P<0.01)。结论:揭示慢性乙型肝炎肝肾阴虚证患者HBV基因突变点的分布规律,有利于进一步探讨中医药抑制HBV突变株复制的方法。     

HBV前C区基因变异与中医证候的相关性研究

研究慢性乙型肝炎患者HBV前C区基因变异的变化规律与中医证候的相关机制。基因芯片检测慢性乙型肝炎不同证候的HBV前C区基因变异。不同位点变异率(%)与证候相关,肝肾阴虚1764(38%),1896(68%),1899(38%);瘀血阻络1762(93%),1764(90%),1899(50%)。不同证候变异株信号强度差异显著,1762、1764;瘀血阻络>肝肾阴虚>肝郁脾虚;1862:肝肾阴虚>肝郁脾虚;>1896:肝肾阴虚>瘀血阻络>湿热中阻>肝郁脾虚;1899:瘀血阻络>肝肾阴虚>湿热中阻>肝郁脾虚。慢性乙型肝炎HBV前C区基因变异与中医证候相关,为分子证候辨证提供了一定实验依据和研究基础。     

乙型肝炎病毒Y(I/V)DD变异与前C及C基因启动子变异生物信息研究

目的 分析HBV反转录酶区(RT)rtM204位点Y(I/V)DD变异与前C及C基因启动子(PC-BCP)变异关系. 方法 应用MEGA4对来自GenBank/EMBL/DDBJ的人类2 849株HBV全基因序列序列重新排列,分析Y(I/V)DD变异与PC-BCP变异及模式相关性. 结果 rtM204(I/V)DD变异株217株(8.0％); PC-BCP变异株1 543株(54.2％),有(1)G1896A、(2)G 1899A、(3) G1896A+G1899A、(4) A1762T/G1764A、(5) A1762T/G1764A+G1896A、(6)A 1762T/G 1764A+G1899A、(7) A1762T/G1764A+G1896A+G1899A 7种变异模式; Y(I/V)DD与PC-BCP联合变异165株.YMDD与PC-BCP双变异率高于单纯YMDD变异率(76％比24.0％,x2=45.283,P=0.000);YIDD与PC-BCP双变异率高于YVDD与PC-BCP双变异率(85％比64.9％,x2=11.836,P=0.000)及使用LAM致YMDD与PC-BCP双变异率高于未使用LAM预存YMDD与PC-BCP双变异率(89.3％比58.9％,x2=27.084,P=0.000).二分类logistic回归分析仅对YIDD有影响而对YVDD无影响的PC-BCP变异模式有G1896A-G1899A(P=0.000,OR=7.573)、A1762T/G1764A-G1899A(P=0.000,OR=6.539)和A1762T/G 1764A-G 1896A-G1899A(P=0.000,OR=6.596). 结论 YMDD变异与PC-BCP变异相关;PC-BCP变异模式的差异与YI/VDD变异选择强度有一定的关系.     

HBV前-C区和BCP区变异与肝细胞癌的相关性分析

目的探讨HBV前-C区G1896A和BCP区A1762T/G1764A变异与肝细胞癌(HCC)的关系及其可能的机制。方法选择于青岛市传染病医院住院的HBV DNA104拷贝/ml的慢性HBV感染者82例,其中慢性乙型肝炎(CHB)29例,乙型肝炎肝硬化(LC)27例,HBV相关肝细胞癌(HCC)26例,采用实时荧光PCR法检测其HBV前-C区1896位变异和BCP区1762/1764位变异,并采用ELISA法检测血清肿瘤坏死因子α(TNF-α)水平。结果 HCC组和LC组前-C区G1896A和BCP区A1762T/G1764A变异率、血清TNF-α水平均显著高于CHB组,但HCC组和LC组间无显著差异。前-C区G1896A和BCP区A1762T/G1764A变异者血清TNF-α水平均显著高于非变异组。结论 HBV前-C区G1896A和BCP区A1762T/G1764A变异与HCC形成有关,机制是否与HBV变异和TNF-α水平间的因果关系相关,有待进一步研究。     

慢性乙型肝炎患者HBV前C区G1896A、核心启动子1762/1764变异与乙型肝炎自然史、血清HBsAg定量及疾病程度的相关性

目的研究HBV前C区G1896A、核心启动子1762/1764变异与HBV自然史、血清HBsAg水平间的相关性及对疾病严重程度的影响。方法分别各选取40例HBV野生株、前C区G1896A或核心启动子1762/1764变异以及两者联合变异的慢性乙型肝炎感染者为研究对象。采用PCR反向点杂交技术检测HBV前C区G1896A位点、核心启动子1762/1764变异及HBV基因分型。同时检测HBsAg定量、HBeAg、HBV DNA定量、肝脏生化指标等。结果(1)HBV前C区G1896A、核心启动子1762/1764变异患者的年龄、ALT高于野生株感染者(P0.001),Alb低于野生株感染者(P0.001)。(2)HBV自然史免疫耐受期以野生株感染为主(P0.001),免疫清除期、低(非)复制期野生株及PC G1896A或/和BCP1762/1764变异株差异无统计学意义(P0.05),再活动期以PC G1896A或/和BCP1762/1764变异株为主(P0.001)。(3)PC G1896A或/和BCP1762/1764变异株的HBsAg水平(lgIu/mL)、HBeAg阳性率较野生株组降低(P0.001),基因C型、HBV DNA水平(≥6 lg拷贝/mL)较野生株组差异无统计学意义(P0.05)。(4)HBV PC G1896A或/和BCP1762/1764变异中肝硬化患者多于野生株(P0.05)。结论慢性HBV感染者前C区G1896A、核心启动子1762/1764变异与乙型肝炎自然史、HBsAg水平、HBeAg的状态及疾病严重程度相关。     

家系感染乙型肝炎病毒基因变异与HLA-Ⅱ类等位基因的关系

目的研究家系内慢性乙型肝炎病毒基因变异特征及与HLA-II类分子基因多态性的关系。方法采用PCR扩增和产物测序检测HBV前C区1896位及BCP区1762/1764位变异,分析家系内慢性HBV感染者121例（实验组）病毒变异特征及与非家系慢性乙肝患者83例（对照组）病毒变异频率差异;用PCR/SSP对家系内慢性HBV感染者的HLA-DQA1和DQB1等位基因多态性进行测定;分析HLA-II类分子基因多态性与家系内慢性乙型肝炎病毒基因变异的关系。结果实验组前C区1896位变异、BCP区1762/1764位双变异及联合变异率均高于对照组;HLA-DQA1＊0501和DQB1＊0301等位基因频率和家系慢性HBV感染者前C区1896位、BCP区1762/1764位及联合变异有关;母亲为第一代的感染家系中,子女的前C区1896位及BCP区1762/1764位变异与母亲是否有该位点变异差异无显著性。结论家系感染HBV患者中,HBV前C区1896位、BCP区1762/1764位双突变及联合变异频率明显高于无家系感染的慢性乙型肝炎患者;家系内感染HBV前C区1896位、BCP区1762和1764位变异并非都是由上一代遗传或传播而来,而主要是与个体的免疫状况有关。     

慢性重型乙型肝炎前C区及BCP区基因突变的临床意义

应用基因芯片杂交技术检测慢性重型乙型肝炎（慢重肝）及非重型肝炎（包括慢乙肝及肝硬化）患者血清中HBV前C区A1896、A1899及BCP区T1762/A1764联合突变的发生率,比较慢重肝好转或治愈患者及恶化或死亡患者前C区及BCP区基因突变情况。结果慢重肝前C区A1896、A1899,BCP区T1762/A1764联合突变率及两区多位点突变率与慢乙肝或肝硬化患者比较均有显著统计学差异（P均〈0.01）;慢重肝好转或治愈患者A1896、A1899及多位点变异率与恶化或死亡患者比较均有统计学差异（P均〈0.05）。提示HBV前C区及BCP区基因变异为慢重肝的重要发病机制之一。     

HBV前C、C、BCP区位点变异与中医证型的相关性分析

目的探讨慢性乙型肝炎(CHB)患者HBV的前C区、C区、基本核心启动子(BCP)区位点变异与中医证型的相关性。方法回顾性分析2014年11月—2018年6月于河南中医药大学第一附属医院脾胃肝胆科门诊及病房诊治的CHB患者临床资料。收集并记录患者的基本情况、HBV血清学指标、HBV各位点变异情况及中医四诊信息,参照病毒性肝炎的中医辨证标准,对各病例进行辨证分型,继而分析HBV前C、C、BCP区位点变异与中医证型的相关性。计数资料组间比较采用χ²检验;计量资料多组之间及两两比较采用Kruskal-Wallis H检验。结果纳入235例CHB患者中,湿热内结证101例,肝郁脾虚证88例,瘀血阻络证17例,肝肾阴虚证19例,脾肾阳虚证10例。性别、年龄、病程在不同中医证型中存在显著差异(χ²=17.389、H=173.280、H=86.520,P值均<0.01)。CHB患者年龄在HBV前C、C、BCP区位点变异间差异有统计学意义(H=30.150,P<0.001)。HBV前C、C、BCP区变异的CHB患者其中医证型分布存在显著差异(χ²=58.117,P<0.001),主要证型为湿热内结证和肝郁脾虚证,两证型所占比例为80.43%。湿热内结证易出现A1762T、G1764A位点变异,肝郁脾虚证易出现G1896A、A1762T、G1764A位点变异,瘀血阻络证和肝肾阴虚证均易出现G1764A位点变异,脾肾阳虚证易出现I97L位点变异。结论CHB患者HBV前C、C、BCP区位点变异与中医证型存在一定的关联性,湿热内结证及肝郁脾虚证最常见,脾肾阳虚证易发生I97L位点变异。     

乙型肝炎病毒基因型、BCP及PC区变异与拉米夫定治疗后HBV DNA反弹的关系

目的:探讨HBV基因型、C区基本核心启动子(BcP)及前C(PC)区变异与拉米夫定抗病毒治疗后HBV DNA反弹的关系.方法:应用多引物对巢式PCR法,PCR-序列分析法,检测拉米夫定治疗27例乙型肝炎患者(治疗组),以及19例从未用过抗病毒治疗患者(对照组)的HBV基因型PC区,BCP的突变位点.结果:27例HBV DNA反弹的患者9例检出G1896A变异率高于对照组(33.33% vs 5.26%,P<0.05),4例检出C1856T变异(14.81%).治疗组4份治疗前标本未检出G1896A、C1856T和BCP变异.与对照组比较,治疗组PC(G1896A)及BCP(A1762T G1764A)双变异的患者中B基因型的构成比增高,分别为75%和50%,C基因型的构成比下降,分别为25%和50%.其中在BCP(A1762T G1764A)变异患者中B、C基因型构成比与对照组比较有显著性差异(P<0.05).4例HBV DNA反弹患者治疗前未检出有基因变异,治疗后有2例检出变异,BCP变异1例,BCP PC变异1例.27例HBV DNA反弹患者BCP变异4例,PC变异2例,BCP PC变异8例.结论:BCP(T1762/A1764)变异、PC区(G1896A)变异可能与拉米夫定治疗后HBV DNA反弹有关.病毒变异导致的HBV DNA反弹可以是单基因变异引起,也可以是多个基因联合变异引起,拉米夫定治疗后B基因型患者更易发生A1762T G1764A变异.     

乙型肝炎病毒前C区和核心启动子区突变与慢性乙型肝炎病情的关系

乙型肝炎病毒（HBV）感染能导致急、慢性肝病，其中全球慢性乙型肝炎感染者近4亿。每年由于HBV慢性感染引起的肝硬化，肝细胞癌死亡患者超过47万人。而由于HBV病毒复制过程缺乏校对机制，在慢性感染的过程中发生基因突变的概率很高，相关研究最多的HBV突变株主要有：发生在翻译水平的前C区突变（G1896A）；发生在核心启动子（BCP）区，即A1762T和G1764A的联合突变；发生在HBV DNA聚合酶区的变异包括P区528、552位点的突变。研究表明，HBV前C区1896位突变以及BCP区突变普遍存在于CHB患者中，     

乙型肝炎病毒前C区和BCP区突变及基因型对HBeAg表达的影响

研究乙型肝炎病毒前C区和BCP区突变及基因型对HBeAg表达的影响。分别用基因芯片和基因测序的方法检验HBV DNA阳性的乙型肝炎病毒前C区和BCP区突变及基因型,应用时间分辨免疫荧光法检测HBeAg含量,按照有无HBV前C区1896、BCP区1762、1764双突变、基因型等指标进行分组分析。无前C区1896和BCP区1762/1764双突变组、单纯1762、1764双突变和1896突变组以及1896、1762、1764联合突变组之间相互比较,HBeAg含量相差显著,F=6.47,P〈0.01;B、C基因型间HBeAg含量相比,相差无显著性,t=0.1394,P〉0.05。乙型肝炎病毒前C区和BCP区突变能显著影响HBeAg量的表达,从而导致乙型肝炎病毒致病能力的变化。     

Mutations in the basic core promotor and the precore region of hepatitis B virus and their selection in children with fulminant and chronic hepatitis B

The involvement of precore stop codon 1896-A and base exchanges in the AT-rich region at positions 1762 and 1764 of the hepatitis B core promotor has been controversely discussed in adults with fulminant hepatitis B. Because no data are currently available on children, we analyzed the basic core promotor (BCP) and precore region in children with chronic and fulminant hepatitis B. The BCP and precore region were sequenced directly and after cloning from mothers and infants. Thirteen children suffered from chronic liver disease, 6 of whom were treated with interferon alfa (IFN-alpha). All 13 patients seroconverted from hepatitis B e antigen (HBeAg) to hepatitis B e antigen antibodies (anti-HBe), and sera were analyzed before and after seroconversion. Nine vertically infected infants developed a fulminant course of hepatitis B. The occurrence of BCP (1762-T/1764-A, 7.7%) and precore (1896-A, 7.7%; 1899-A, 15%) mutations in chronic hepatitis B was rare. A genotype shift from D to A was observed in 3 patients after development of anti-HBe. A high number of base exchanges was detected in those infants with fulminant hepatitis B. Eight of nine showed a G-A exchange at positions 1896/97 (89%), 1899 (56%), and/or mutations at nucleotide (nt) positions 1762 (56%) and 1764 (78%). All virus strains belonged to genotype D, whereas in the only surviving infant, a D-to-A shift was detected. Hepatitis B virus (HBV) DNA clones were examined from 3 babies and 5 mothers. Our results showed a heterogeneous virus population in 4 of 5 mothers. In contrast, a homogeneous virus population emerged in the infants. According to our data, the analysis in children with fulminant and chronic hepatitis B revealed a striking presence of BCP and precore mutants in infants with fulminant hepatitis (FH) when compared with clinically inapparent anti-HBe-positive children (P <.002), which could be one factor in the pathogenesis of fulminant hepatitis B in children.     

慢性重型乙型肝炎患者HBVDNA前C／BCP区突变基因分析

目的分析慢性重型乙型肝炎（慢重乙肝）患者HBVDNA前C区和基本核心启动子（前C／BCP）区突变特点与意义。方法收集87例慢重乙肝和196例慢性乙型肝炎（慢乙肝）患者血清，提取HBVDNA，用巢式PCR扩增HBVDNA前C／BCP区基因，PCR产物进行DNA测序，用NBI软件比对结果，重点分析G1896、G1862、G1899、A1762、G1764、T17536个位点突变。结果慢重乙肝组和慢乙肝组6个位点突变全阴率分别为3．4％和28．1％（P〈0．01）；慢重乙肝组在其中5个位点上的突变检出率显著高于慢乙肝组。此外，慢重乙肝组和慢乙肝组≥三联突变检出率分别为56．3％和35．2％（P〈0．01），≥四联突变检出率分别为25.3％和8．7％（P〈0．01），插入／缺失突变检出率分别为10．3％和1．0％（P〈0．01）。结论HBVDNA前c／BcP区基因突变发生频率的增加与慢乙肝发生重症化相关，结合临床资料分析突变的意义将有助于认识慢乙肝重症化的发生机制。     

乙型肝炎病毒基因组变异对干扰素治疗的影响

目的 研究乙型肝炎病毒基因组Ｘ区和前Ｃ／Ｃ区多部位变异对干扰素（ＩＦＮ）治疗的影响。方法 利用套式聚合酶链反应产物直接测序技术，测定１７例慢性乙型肝炎患者ＩＦＮ治疗前病毒核苷酸序列。结果 ５例Ｔ＾１７６２－Ａ＾１７６４变异株感染者均对ＩＦＮ治疗有应答，而在１２例无Ｔ＾１７６２－Ａ＾１７６４突变患者中仅３例有应答。在８例Ａ＾１８９６变异株感染者中，５例同时伴Ｃ区Ｂ细胞表位ａａ１０７￣１１８突变，其中     

Role of hepatitis B virus base core and precore/core promoter mutations on hepatocellular carcinoma in untreated older genotype C Chinese patients

The aim of the study was to investigate the prevalence of mutations of basal core promoter (BCP) and precore (PreC) region of hepatitis B virus (HBV) and their association with hepatocellular carcinoma. A total of 341 untreated older HBV patients were divided into three groups: chronic hepatitis B (CHB, 185), cirrhotic hepatocellular carcinoma (LC-HCC, 113) and non-cirrhotic hepatocellular carcinoma (non-LC-HCC, 43). HBV BCP and PreC mutations and genotypes were determined by direct sequencing. Using univariate analysis, age (≥ 45 years), single mutations including A1896 and A1899 and multiple mutations T1762/A1764 + A1896, T1762/A1764 + A1899 and T1762/A1764 + A1896 + A1899 were more frequently detected in LC-HCC and non-LC-HCC patients than in CHB patients. BCP T1762/A1764 mutations were highly detected in LC-HCC patients than in CHB patients. Multivariate logistic regression analysis (adjusted for age and gender) revealed that among HBeAg-positive patients, BCP T1762/A1764 mutations (OR, 5.975; P = 0.05), PreC A1899 mutation (OR, 4.180; P = 0.013) and multiple mutations T1762/A1764 + A1899 (OR, 6.408; P = 0.006) were independently associated with the development of LC-HCC; PreC A1899 mutation (OR, 7.347; P = 0.034) was also independently associated with the development of non-LC-HCC. On the other hand, among HBeAg-negative patients, PreC A1896 mutation (OR, 5.176; P = 0.002) and multiple mutations T1762/A1764 + A1896 (OR, 4.149; P = 0.007) were independently associated with the development of non-LC-HCC. These results indicated that older age (≥ 45 years) was associated with LC-HCC and non-LC-HCC development. BCP T1762/A1764 mutations and PreC A1899 mutation were associated with the LC-HCC development in HBeAg-positive patients. PreC A1896 mutation was associated with the non-LC-HCC development in HBeAg-negative patients.     

Incidence and clinical significance of hepatitis B virus precore gene translation initiation mutations in e antigen-negative patients

Hepatitis Be antigen (HBeAg)-negative chronic hepatitis B (CHB) is associated with hepatitis B virus (HBV) variants harbouring changes in the precore region. Most commonly, a G to A point mutation at nucleotide 1896 (m1896) creates a novel translation stop codon that prevents HBeAg production. In the Mediterranean region the m1896 mutation prevails in greater than 98% of HBeAg-negative CHB patients. In this study the prevalence of additional mutations in the precore region was investigated among patients with chronic HBV infection. Precore sequences were determined by sequencing serum HBV DNA amplified by polymerase chain reaction (PCR) with primers flanking the precore/core region. Thirty-one HBeAg-negative and five HBeAg-positive individuals were studied. All HBeAg-negative patients (100%) harboured the m1896 mutation and 20 (64.5%) also had a G to A mutation at nucleotide 1899 (m1899). Additional mutations affecting the translation initiation of the precore gene were found in seven (22.5%) patients, all with active liver disease, five of whom had episodes of HBV reactivation. HBeAg-positive patients had no mutations in these positions and neither did any of the five HBeAg-negative patients with normal levels of liver enzymes, representing the healthy carrier state of HBV infection. Serial sample analysis from one patient revealed that the initiation codon mutation developed following HBeAg seroconversion and the appearance of m1896. During periods of high HBV replication, the ratio of mutant to wild-type ATG was found to increase in parallel with HBV DNA levels. These data show that a significant proportion of HBeAg-negative patients who already harbour the 1896 stop codon mutation may subsequently develop precore translation initiation mutations, which appear to be associated with enhanced HBV replication and severe liver disease.     

Detection of HBV core promoter and precore mutations helps distinguish flares of chronic hepatitis from acute hepatitis B

Background and Aim: Acute exacerbation of chronic hepatitis B has to be distinguished from acute hepatitis, because treatment strategies differ between them. Methods: Mutations in the core promoter and precore region of hepatitis B virus (HBV) were determined in 36 patients with acute exacerbation of chronic hepatitis B, in whom alanine aminotransferase (ALT) increased above 500 IU/L, as well as the 36 patients with acute hepatitis. Results: Mutations in the core promoter (A1762T/G1764A) and precore region (G1896A) were more frequent in patients with acute exacerbation of chronic hepatitis than acute hepatitis (81% vs 19%; P < 0.0001 and 58% vs 6%; P < 0.0001, respectively). Of the 19 patients with mutations in both the core promoter and precore region, 17 (89%) had acute exacerbation of chronic hepatitis. In contrast, among the 32 patients with the wild‐type for both the core promoter and precore region, 29 (89%) developed acute hepatitis. By multivariate analysis, the double mutation in the core promoter was predictive of acute exacerbation in chronic hepatitis with the highest odds ratio at 26.4. Conclusions: In patients with hepatitis B having ALT levels >500 IU/L, mutations in the core promoter and precore region are useful in distinguishing acute exacerbation of chronic from acute HBV infection. Detection of these mutations would be useful for commencing prompt antiviral treatments on patients with acute exacerbation of chronic hepatitis for a better prognosis.     

重型乙型肝炎e抗原阴性患者前C变异株及其体外翻译

目的：探讨HBeAg阴性的慢性重型乙型肝炎患者HBV前C区变异及前C区T1862突变体对e抗原合成和分泌影响,方法：选取9例重型乙型重病毒性肝炎患者,用PCR方法扩增血清中HBV前C／C基因区,并克隆后测序和序列分析,构建HBV前C区T1862位点突变体表达质粒pGEMT,经体外翻译比较野毒株和变异株的表达产硪,结果：HBV前C区有3个位点异使氨其酸序列改变,A1896,A1899和T1862,A1899并不单独出现,前C区T1862突变并不阻止HBVe前体外合成,同时,有2例并未检测到前C区的变异,结论：部分重型肝炎HBeAg阳性原因降了T1896变异外,还存在T1862变异,前C1862突变对HBVe抗原前体蛋白合成并无影响,可能e抗原前在分泌过程中受阻。     

Association of core promoter/precore mutations and viral load in e antigen-negative chronic hepatitis B patients

Apart from core promoter A1762T/G1764A and precore G1896A mutations, other hepatitis B virus (HBV) mutants are detected in hepatitis B e antigen (HBeAg)-negative chronic hepatitis B (CHB). The aim of this study was to determine the effects of those mutants on clinical manifestation and viral loads of genotypes B and C HBV. Seventy-nine HBeAg-negative CHB patients with hepatitis flare were enrolled in this study and their HBV precore/core region were sequenced. Serial biochemical profiles and viral loads were assessed and compared. Fifty-three patients (67%) were infected by genotype B HBV and 26 (33%) were infected by genotype C HBV. The clinical manifestation and HBV viral loads were comparable between the two groups. However, genotype B was significantly associated with precore G1896A mutation (92.5%), and more mutations within nucleotide 1809-1817 were detected in patients infected by genotype B as compared with those infected by genotype C (18.9%vs 3.8%). Most of the cases had mutations at the -2, -3 or -5 position from the precore AUG initiation codon. Triple core promoter mutations T1753C/A1762T/G1764A [corrected] appeared to be linked to genotype C rather than genotype B HBV (19.2%vs 1.9%; P = 0.013). In multivariate analysis, the presence of either triple core promoter 1753/1762/1764 mutation or nucleotide 1809-1817 mutation was the only factor associated with lower HBV viral load (<70 Meq/mL) (odds ratio = 9.01; 95% CI 1.11-71.43; P = 0.04). In conclusion, minor HBV variants with mutations in the core promoter and precore region were detectable in genotypes B and C. Such HBV variants are genotype specific and related to viraemia levels.