慢性乙型肝炎病毒感染者病毒前C区和基本核心启动子区变异检测及意义

目的 探讨乙型肝炎病毒（HBV）前C区和基本核心启动子（BCP）区变异与基因型及疾病进展间的关系。方法 收集HBV携带者（ASC）、慢性乙型肝炎（CHB）、肝炎肝硬化（LC）、肝细胞肝癌（HCC）患者血清148份，用半巢式聚合酶链反应扩增HBV前C／C基因部分片段，产物纯化后直接测序，检测前C区A1896及BCP区T1762／A1764变异。用S基因聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）方法确定HBV基因型。结果 有128份血清能够成功分型和测序，其中B基因型60份，C基因型68份。在B基因型感染者中前C区A1896变异检出率（48．33％）明显高于C基因型感染者（29．41％，X^2=4．83，P〈0．05）；而BCP区T1762／A1764变异检出率却明显低于C基因型感染者，差异亦有统计学意义（30．00％：73．54%，X^2=24．25。P〈0．05）。前C区A1896变异在CHB、LC、HCC中的阳性检出率分别为46．88％（15/32）、39．39％（13／33）、51．52％（17／33）。与ASC的13．33％（4／30）相比，P分别〈0．05，差异有统计学意义。BCP区T1762／A1764变异检出率在HCC、LC组分别为87．88％（29／33）和72．73％（24／33）．明显高于CHB组的37．50％（12／32）及ASC组10.00％（3／30）（P〈0．05）。结论 前C区A1896变异常见于B基因型感染者，而BCP区T1762/A1764变异C基因型感染者多见。除ASC外．前C区A1896变异与疾病进展关系不大．而BCP区T1762/A1764变异与乙型肝炎进展及顶后相关。     

乙型肝炎病毒基因型、YMDD变异与拉米夫定治疗后HBV DNA反弹关系的研究

目的探讨HBV基因型、YMDD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的27例乙型肝炎患者和19例从未用过抗病毒治疗的患者HBV基因型和P区（YMDD）的突变位点。结果在27例HBV DNA反弹的患者中，13例（48．15％）检出YMDD变异，而对照人群无YMDD变异（P〈0．05）。YMDD变异的位点为rtM204V／I（C区）±rtL180M（B区）；在治疗组YMDD变异的患者中，B、C基因型构成比（46．15％和59．26％）与对照组（53．85％和68．42％）比较无显著性差异（P〉0．05）。结论YMDD变异是拉米夫定治疗后出现耐药导致HBV DNA反弹的主要原因；YMDD变异的常见位点依然为rtM204V／I（C区）±rtL180M（B区）；YMDD变异在B、C基因型病人中无差别。     

拉米夫定耐药株感染者HBV基因型和逆转录酶区变异分析

目的了解拉米夫定耐药株感染者HBV基因型特征并分析耐药株HBV逆转录酶（RT）区变异位点和变异类型。方法应用PCR扩增和直接测序HBV逆转录酶区并与Genebank中90株不同基因型野毒株序列进行比较,确定54例耐药株感染者HBV基因型和HBVRT核苷酸的变异特点。结果在54例拉米夫定耐药株感染者中,HBVB基因型占27．78％,C型占70．37％,B／C混合型占1．85％;51例患者出现RT保守区氨基酸变异（包括550和526位氨基酸变异）;18例患者出现除主型区外HBVRT非主型区伴随变异;3例患者未检测到与拉米夫定相关性变异。结论拉米夫定耐药株感染者HBV基因型主要为B型和C型;拉米夫定耐药株的氨基酸变异不仅见于RT区的526和550两个位点,其他位点以及RT非保守区也可发生变异。     

乙肝患者拉米夫定治疗过程中YMDD耐药突变株的变化

目的探讨乙肝患者拉米夫定治疗过程中YMDD变异与HBV基因型、HBV DNA含量及治疗时间的关系。方法采用实时PCR法和基因测序法分别对107例拉米夫定治疗的慢乙肝患者进行HBV DNA含量和HBV基因分型、YMDD变异检测。结果 107例慢乙肝患者中,B型19例（17.8%）,C型81例（75.7%）,B、C混合型7例（6.5%）,未发现A、D、E、F、G、H等其他基因型;共25例（23.4%）发生YMDD变异,其中B型、C型和B、C混合型YMDD变异的发生率分别为26.3%、22.2%、28.6%。各基因型患者YMDD变异的发生率及Y IDD、YVDD的变异类型均无统计学差异（P〉0.05）。血清HBV定量水平低、中、高度组产生YMDD变异株的时间有统计学差异（P〈0.05）。结论 YMDD变异与HBV基因型无明显相关性;治疗前血清HBV DNA含量越高,拉米夫定治疗期间发生YMDD变异的时间越早;出现YMDD变异患者肝组织内处于不同的炎症及纤维化状态。     

慢性乙型肝炎病毒基因型与拉米夫定疗效关系的研究

目的：研究乙型肝炎病毒(HBV)基因型对拉米夫定抗病毒疗效的影响。方法：回顾性调查286例拉米夫定治疗组和对照组患者的临床资料。结果：HBV优势基因型是B(34％)和C(48％)型，共235例。135例患者接受拉米夫定抗病毒治疗，对照组100例。拉米夫定有效率在B、C两基因型中分别为92．9％和75．9％(P=0．02)，而对照组分别为9．8％和8．5％(P=0．59)；YMDD变异发生率为8．9％和22．8％(P=0．028)。Multivariate分析发现，B基因型、ALT升高、HBV DNA低水平是影响抗病毒应答的预测因素。在ALT升高患者中，B、C基因型拉米夫定有效率分别为93％和77％(P=0．01)，对照组为13％和8％(P=0．45)。Multivariate分析发现，B基因型、HBV DNA低水平是预测较好疗效的独立因素。结论：B基因型HBV对拉米夫定的应答率高于C型，而变异YMDD发生率低于C型，基因型是影响拉米夫定疗效和诱导变异的重要因素之一。     

乙型肝炎病毒基因型与YMDD变异的关系

目的:了解乙型肝炎病毒(HBV)基因型与YMDD变异之间的关系.方法:多对型特异性引物PCR扩增法对238例经拉米夫定治疗的慢性乙型肝炎患者进行HBV基因分型,直接序列分析;采用基因芯片检测YMDD及前C/BCP区变异.结果:238例患者中,检测出B基因型190例(79.8%),C基因型41例(17.2%),BC混合型7例(3.0%);发生YMDD变异44例,变异率为18.5%,其中B基因型33例,变异率为17.4%,C基因型8例,变异率为19.5%,BC混合型3例,变异率为42.4%,C基因型YMDD变异的发生率与B基因型相比,差异无显著性意义(P＞0.05).44例YMDD变异者中,30例同时存在L528M变异,7例联合前C区(nt 1 896)变异,13例联合BCP区(nt 1 762/1 764)双重突变.结论:本地区慢性乙型肝炎患者中,优势基因型为B型和C型,经拉米夫定治疗后YMDD变异的发生率在B型和C型差异无显著性意义,同时伴有L528M及前C/BCP区多重变异.     

拉米夫定治疗前后乙型肝炎病毒YMDD变异的相关因素分析

目的 了解遵义地区HBV基因型以及拉米夫定治疗前后发生YMDD变异的相关因素,及早进行拉米夫定疗效及耐药的预测. 方法 53例慢性乙型肝炎患者分别在口服拉米夫定前及治疗后3、6、12、18、24个月进行血清HBV DNA定量、乙型肝炎标志物、ALT、AST、总胆红素,白蛋白的检测.同时在接受拉米夫定治疗前采用基因测序法检测HBV基因型及YMDD变异株,治疗后HBV DNA定量下降又反弹升高,且血清HBV DNA＞1×104拷贝/ml时,再次进行YMDD变异株检测.率的比较用卡方检验及确切概率法,两组均数之间比较采用独立样本t检验,有序变量之间的比较采用秩和检验.结果 遵义地区的HBV基因型由B、C及B+C基因型构成.拉米夫定治疗后18例检出YMDD变异株,用药1年和2年的变异率分别为15.1%和34.0%.HBV突变类型有rtL180M/M204V、rtL180M/M204I、rtM204I和rtL180M四种,其中C区rtM204V全部合并rtL180M突变(100%),C基因型中rtL180M/M204V联合突变及rtL180M/M204I联合突变明显高于B基因型(77.8%比25.0%及22.2%比12.5%);C基因型中未见点突变,而rtM204I、rtL180M的点突变仅见于B基因型.YMDD变异与未变异组性别、民族、乙型肝炎家族史及HBeAg情况差异无统计学意义(P＞0.05),病程≥2年组和年龄＜35岁组变异率明显升高(X2值分别为4.707和5.853,P值均＜0.05).不同HBV DNA滴度患者YMDD变异率差异无统计学意义(X2=0.801,P＞0.05),但HBV DNA＜105拷贝/ml者未发现YMDD变异.结论 拉米夫定治疗后YMDD变异可能与HBV基因型及P基因突变类型有关,并随治疗时间的延长而增加.为了减少YMDD变异的发生,应选用病程短、HBV DNA水平较低、肝损害较重的患者进行拉米夫定治疗,有条件的应检测HBV基因型.     

拉米夫定对湖北地区乙型肝炎病毒基因型的影响

目的研究拉米夫定对湖北地区乙型肝炎病毒基因型的影响及其临床意义。方法采用多对型特异性引物-聚合酶链反应检测160例慢性乙型肝炎患者血清HBV基因型;采用基因芯片技术对86例拉米夫定治疗18个月的慢性乙型肝炎患者进行酪氨酸—蛋氨酸—天冬氨酸—天冬氨酸(YMDD)基序、G1896A、A1814C、A1792T和G1764A单碱基变异检测。结果160例慢性乙型肝炎患者,B基因型127例(79%),C基因型24例(15%),BC混合型9例(6%),未发现A、D和E基因型。拉米夫定治疗的86例患者中,17例(19.7%)发生YMDD变异,B型14例,C型2例,BC混合基因型1例,其中6例发生多重变异,包括B型4例,C型2例;HBeAg/HBeAb血清转换率B型41例(68.3%)高于C型8例(40%)(P<0.05)。另外17例患者,经拉米夫定治疗后,HBVDNA仍阳性,亦未发现YMDD变异株。结论湖北地区HBV存在B、C和BC混合基因型,B型为本地区优势基因型,B型在拉米夫定治疗中更易发生YMDD变异;未变异者,血清HBeAg/HBeAb转换率高。     

HBV基因型及P基因变异与拉米夫定耐药的关系

为探讨拉米夫定耐药与HBV基因型和P基因变异的关系,用基因测序的方法分析基因型与YMDD变异,用荧光定量的方法,检测HBVDNA定量,研究结果表明C基因型比其它基因型更易耐药,耐药的问题不仅仅是 YMDD变异,P基因可能诱导其它基因的变异,导致拉米夫定抗病毒治疗的疗效减低。     

慢性乙型肝炎病毒感染者病毒YMDD的自然变异

目的 了解慢性HBV感染患者外周血HBV YMDD自然变异情况及其影响因素.方法 采用引物特异性实时荧光PCR法检测慢性HBV感染者外周血HBV YMDD变异情况,并对影响YMDD自然变异检出率的可能因素进行单因素及多因素分析.根据不同资料分别采用χ~2检验、Fisher's确切概率法、t检验、秩和检验及Logistic回归分析进行统计学处理. 结果在196例未经抗病毒治疗的慢性HBV感染者中,检出存在YMDD自然变异株感染者21例(10.70%),其中YVDD阳性20例,YIDD阳性例1变;变异毒株占总病毒株超过50%者1例,25%～500者5例,9%～25%者15例.B基因型HBV感染病例中YMDD变异株的检出率(20.00%,12/60)显著高于C基因型HBV感染病例(7.38%,9/122),χ~2=6.28,P<0.05.患者性别、年龄、HBeAg状态、HBVDNA载量、疾病状态、病毒感染时间对YMDD自然变异株的检出率无显著影响. 结论 在未经抗病毒治疗的慢性HBV感染者中存在HBV YMDD自然变异;YMDD自然变异的发生率与患者性别、年龄、HBeAg状态,HBV DNA载量、疾病状态、感染时间无显著相关性.B基因型较C基因型HBV更易出现YMDD自然变异.     

乙型肝炎病毒YMDD及e抗原相关多重变异及其临床意义

目的 研究拉米夫定治疗慢性乙型肝炎期间HBVYMDD基序、影响HBeAg分泌的多重变异情况与临床的关系。方法 采用基因芯片技术对拉米夫定治疗9～30个月的慢性乙型肝炎患者进行YMDD基序、G1896A、A1814C、A1762T和G1764A(BCP双突变)单碱基变异检测。结果 102例慢性乙型肝炎患者拉米夫定平均治疗18个月时,22例发生YMDD变异,其中8例发生多重变异,包括G1896A3例、A1814C2例、G1896A A1814C、BCP双突变、BCP双突变 G1896A多重变异各1例,单纯YMDD变异和前5例联合变异均为HBeAg阳性,而后3例多重变异则为HBeAg阴性,其中1例多重变异继续治疗3个月后转变为单纯YMDD野生株阳性,同时伴有HBeAg的复阳。结论 拉米夫定治疗过程中存在YMDD及HBeAg相关多重变异的优势病毒株可能是HBVDNA复阳、同时伴有HBeAg阴转的原因之一,拉米夫定治疗过程中,HBeAg阴性时应监测其可能的相关变异。     

High level of hepatitis B virus DNA after HBeAg-to-anti-HBe seroconversion is related to coexistence of mutations in its precore and basal core promoter

AIM: G1896A mutation in precore or A1762T/G1764A mutations in basal core promoter are suspected to be responsible for patients with detectable level of HBV DNA in serum after seroconversion from HBeAg to anti-HBe. However, G1896A variant has impaired, while A1762T/ G1764A variant may have intact replication ability. They themselves or their coexistence status may play different roles in such meaningless seroconversion. For these reasons, the significances of these two types of mutations were comparatively investigated in this study. METHODS: One hundred and sixty-five sera with positive anti-HBe and HBV DNA were collected from different patients. Mutations of G1896A and A1762T/G1764A among these serum samples were detected using competitively differentiated PCR. HBV DNA was demonstrated using real-time quantitative PCR. RESULTS: G1896A and/or A1762T/G1764A mutations were detected in 89.1% (147/165) out of patients with detectable HBV DNA in serum after HBeAg-to-anti-HBe seroconversion. The positive rate of G1896A variants was significantly higher than that of A1762T/G1764A mutations (77.6% vs 50.3%, X2= 26.61,P<0.01). The coexistence positive rate of these two types of mutations was 38.8% (64/165). Coexistence mutations were found in 77.1% (64/83) out of sera with A1762T/G1764A mutations, and in 50.0% (64/128) out of sera with G1896A mutation. Compared with variants with G1896A mutation only, the coexistence mutations were predominant in patients with high level of serum HBV DNA, and related to higher total bilirubin, lower serum albumin and progressive liver diseases. CONCLUSION: The coexistence of G1896A mutation and A1762T/G1764A mutations is very common, and responsible for the major cases with high level of HBV DNA in serum and progressive liver diseases after HBeAg-to-anti-HBe seroconversion. This coexistence mutation variant may have higher pathogenicity and replication ability.     

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.     

Clinical significance and evolution of core promoter and precore mutations in HBeAg-positive patients with HBV genotype B and C: a longitudinal study.

BACKGROUND/AIMS: The aims of this longitudinal study were to investigate whether the clinical outcome and evolution of core promoter and precore mutations were different during hepatitis B e antigen (HBeAg) seroconversion between hepatitis B virus (HBV) genotypes B and C in HBeAg-positive patients with chronic hepatitis B. PATIENTS AND METHODS: The core promoter and precore sequences were determined from serial sera of 156 HBeAg-positive patients with chronic HBV infection. RESULTS: In HBV genotype C, the T1762/A1764 mutant was detected earlier than the A1896 mutant, and the frequency was significantly higher than in HBV genotype Ba over the entire follow-up period. In HBV genotype Ba, A1896 was found earlier than the T1762/A1764 mutant, and the frequency was significantly higher than in genotype C only before HBeAg seroconversion, and the A1896 mutant played an important role in HBeAg seroconversion in HBV genotype Ba. In addition, the T1846 variant was an independent factor associated with HBeAg seroconversion. Furthermore, HBV genotype C was associated with the development of G or C1753 and T1766/A1768 mutations, and the reactivation of hepatitis after HBeAg seroconversion. Based on Cox's regression analysis, the significant risk factors of liver cirrhosis were older age at entry [hazard ratio (HR)=1.085, 95% confidence interval (CI)=1.036-1.136, P=0.001], alanine transaminase (ALT) >80 U/l (HR=3.48, 95% CI=1.37-8.86, P=0.009), and the T1762/A1764 mutant (HR=5.54, 95% CI=2.18-14.08, P<0.001). CONCLUSIONS: Our study showed that different HBV genotypes were associated with various mutations in the core promoter and precore regions during HBeAg seroconversion. T1762/A1764 mutation could be useful in predicting clinical outcomes in HBeAg-positive patients with HBV infection.     

HBeAg阴性慢性乙型肝炎患者出现前C和基本核启动子联合突变的意义

目的 探讨HBeAg阴性慢性乙型肝炎患者出现前C区G1896A突变和基本核启动子(BCP)区A1762T／G1764A双突变，以及两者的联合突变对患者血清病毒含量的影响。方法 收集240份HBeAg阴性、60份HBeAg阳性慢性乙型肝炎患者血清及40份阴性对照血清，采用竞争分化聚合酶链反应(CD-PCR)检测G1896A突变及A1762T/G1764A双突变，采用实时荧光定量聚合酶链反应(PCR)检测血清中病毒含量。结果 G1896A突变在HBeAg阴性和HBeAg阳性患者中的检出率分别为57．6％和6．7％；A1762T／G1764A双突变的检出率分别为37．9％和31．7％；其中两者联合突变在HBeAg阴性患者中的检出率为13．5％。在HBeAg阴性患者中，G1896A突变主要出现在血清病毒含量低的患者，而A1762T／G1764A双突变与血清病毒含量无关。联合变异株主要见于重度慢性乙型肝炎患者，与血清病毒含量无关。结论 G1896A变异株复制能力较低，而A1762T／G1764A变异株对病毒复制能力影响可能较为复杂，对HBeAg合成的影响较G1896A变异株小。联合变异株的致病力相对较强，其复制能力较单纯G1896A强，值得警惕。     

福州市乙肝病毒基因型和基因亚型的分布及其与前C、C基因启动子变异的关系

目的 研究福州市乙肝病毒基因型和亚型分布及其与T1762/A1764、A1896变异的关系,为完善预防、诊断、治疗病毒感染的策略和方法提供科学依据. 方法 应用型特异性引物PCR法检测HBsAg阳性血清的基因型,应用PCR-RFLP方法检测基因亚型、T1762/A1764变异和A1896变异. 结果 282份HBsAg阳性血清样品中103份未能成功分型,其余179份样品中B基因型122份(68.2%),C基因型54份(30.2%),B C型3份(1.7%),未检测到其他基因型.随机选取的100份B基因型样品中,Ba亚型71份(71.0%),Bj亚型8份(8.0%),未能分亚型者21份(21.0%).54份C基因型样品中Ce亚型 31份(57.4%),Cs亚型 14份(25.9%),Ce Cs 1份(1.9%),未能分亚型者8份(14.8%).T1762/A1764变异标本9份(8.7%),Ce亚型变异率最高(29.2%),Ba亚型次之(3.3%),Cs和Bj亚型未检测到变异株,T1762/A1764变异在不同基因型和亚型间的分布差异有统计学意义(P＜0.05).A1896变异标本10份(10.0%),Ba亚型变异率最高(14.0%),Cs亚型次之(10.0%),Ce亚型最低(4.0%),不同基因型和亚型中的变异差异无统计学意义(P＞0.05).HBeAg阳性和阴性样品中的基因型和亚型分布差异无统计学意义(P＞0.05). 结论 福州市乙肝病毒以B、C基因型为主,Ba、Ce亚型占优势,HBV各基因型和亚型发生T1762/A1764、A1896变异的模式不同.     

三种基因亚型乙型肝炎病毒感染者的临床资料与病毒学指标差异分析

目的 调查HBV Ba、C1和C2三种基因亚型在临床致病性方面的差异,同时检测并分析三种基因亚型毒株在前C和C基因启动子区发生变异的模式.方法 采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)法对来自广东地区151例慢性HBV感染者血清进行HBV基因亚型分析,同时测定所有标本前C及C基因启动子区的序列.结果 151份样本中Ba亚型占53.0%(80/151),C1亚型占33.8%(51/151),C2亚型占13.2%(20/151).三种亚型的患者在年龄和性别构成比无差异的情况下,其HBeAg阳性率以及多项肝功能指标差异均无统计学意义,但在前C及C基因启动子区的变异模式上存在明显差异:Ba亚型易发生A1896终止密码变异,T1762/A1764变异的发生概率较低;C1亚型易发生T1762/A1764变异,A1896变异发生概率较低;C2亚型发生T1762/A1764和A1896变异的概率介于Ba和C1亚型之间.结论 三种HBV基因亚型毒株可能采取不同的变异模式形成HBeAg阴性HBV感染.     

Hepatitis B virus genotype B and mutations in basal core promoter and pre-core/core genes associated with acute-on-chronic liver failure: a multicenter cross-sectional study in China

Background and aim

In China, acute-on-chronic liver failure (ACLF) is mostly caused by hepatitis B virus (HBV). However, the mechanism remains unclear. This study aims to investigate the association between both HBV genotype and mutations in basal core promoter (BCP) and pre-core/core (pre-C/C) regions with the development of HB-ACLF.

Methods

A multicenter cross-sectional study was performed in China. Serum samples from 522 patients were analyzed, including 231 patients with mild-chronic hepatitis B (CHB-M), 84 with severe-chronic hepatitis B (CHB-S) and 207 with HB-ACLF. HBV genotype and related mutations in the BCP and pre-C/C regions were determined by direct sequencing.

Results

A significantly higher ratio of HBV genotype B to C was detected in HB-ACLF patients than in CHB-M or CHB-S patients. The A1762T/G1764A, A1846T and G1896A mutations were significantly more common in HB-ACLF patients infected with either genotype B or C as compared with CHB-M, whereas the C1913A/G and A2159G mutations were more associated with HB-ACLF in genotype C patients. Comparing with CHB-S, the A1762T/G1764A mutation in genotype B and the A2159G mutation in genotype C were significantly more common in HB-ACLF patients. A multivariate analysis showed that factors such as HBV genotype B, age ≥40 years and A1762T/G1764A, A1846T and G1896A mutations were independently associated with the development of HB-ACLF.

Conclusion

Chronic HBV infection with genotype B, A1762T/G1764A, A1846T and G1896A mutations has a higher possibility to develop HB-ACLF. These virological factors could serve as possible molecular markers for prediction of the clinical outcomes of chronic HBV infection.     

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。     

Real-time quantification of hepatitis B virus core-promoter and pre-core mutants during hepatitis E antigen seroconversion

BACKGROUND/AIMS: Detection of hepatitis B virus (HBV) core-promoter A(1762)T-G(1764)A and pre-core G(1896)A mutants has relied on qualitative assays. We tested the hypothesis that the quantity of A(1762)T-G(1764)A and G(1896)A mutants might have clinical impact, by quantifying these mutants before and after HBe antigen (HBeAg) seroconversion in 58 patients. METHODS: A real-time quantitative-polymerase chain reaction (Q-PCR) was developed, using minor groove binder (MGB)-conjugated TaqMan probes to impart reaction specificity for wildtype/mutant HBV populations. RESULTS: Significant quantities (>20%) of core-promoter A(1762)T-G(1764)A mutant existed in 65% of patients before and after HBeAg seroconversion, and were significantly changed (>20% increase/decrease) in 13% of patients after seroconversion. Quantity of A(1762)T-G(1764)A mutants was positively correlated with alanine aminotransferase (ALT) (P<0.001) and HBV DNA (P<0.001) levels, both before and after HBeAg seroconversion. Significant quantities of pre-core G(1896)A mutant existed in about 90% of patients before and after HBeAg seroconversion, and were changed in 16% of patients after seroconversion. Quantity of G(1896)A mutant was negatively correlated with ALT (P=0.044) and HBV DNA (P=0.007) levels. CONCLUSIONS: The A(1762)T-G(1764)A and G(1896)A mutants existed in a high proportion of patients before and were unaffected after HbeAg seroconversion. The quantities of A(1762)T-G(1764)A mutant were positively and G(1896)A mutant negatively correlated with liver inflammation and viral replication.