乙型肝炎病毒前核心区及基本核心启动子变异的检测及其对患者疾病谱的影响

目的探讨乙型肝炎病毒(HBV)前核心区(前C区,nt1896)及基本核心启动子(BCP,nt1762/1764)变异在慢性HBV感染者疾病谱的分布及对患者疾病谱的影响。方法416例血清HBsAg阳性、HBVDNA定量大于1.0×104拷贝/毫升的患者,采用微流基因芯片检测HBV前C区及BCP变异。结果416例HBV感染者中302例为HBeAg(-)患者,其中248例(82.12%)有前C区或BCP变异,41.06%为前C区变异,31.12%BCP变异,2种同时变异为9.94%。HBeAg(-)的慢性乙型肝炎和肝硬化患者前C区变异分别为64.72%和83.33%(x2=0.89,P>0.05),均大于HBeAg(-)无症状携带者的28.47%(x2=54.20,P<0.01;x2=5.29,P<0.05);而HBeAg(-)无症状携带者BCP变异达77.19%,大于HBeAg(-)慢性乙型肝炎和肝硬化患者(x2=69.73,P<0.01;x2=10.58,P<0.01)。而在114例HBeAg(+)患者中28.95%有前C区或BCP变异。结论前C区或BCP变异在慢性HBV感染者疾病谱的分布不同,在HBeAg(-)/HBVDNA(+)与HBeAg(+)/HBVDNA(+)患者变异率差异显著。HBV前C区可能是该病变反复及加重的一个重要原因,但BCP变异临床意义不明确。     

抗-HBe阳性乙型肝炎病毒C基因启动子和前C基因变异

研究抗 -HBe阳性乙型肝炎病毒C基因启动子 (CP)和前C基因变异。用基因序列分析检测基因变异。(1)抗 -HBe阳性患者的前C终止变异 (nt1896G→A)的发生率显著高于抗 -HBe阴性组 (P <0 0 0 1) ;而CP双变异(nt176 2A→T和 176 4G→A)则在两组中无明显差异。 (2 )同抗 -HBe阳性慢性乙肝组比较 ,抗 -HBe阳性重型乙肝患者的前C终止变异和CP双变异发生率无明显差异 ,而CPnt175 2A→G变异发生率则显著增高 (P <0 0 5 )。前C终止变异与抗 -HBe阳性乙型肝炎密切相关 ;而CPnt175 2变异则可能与抗 -HBe阳性重型乙型肝炎发病有关     

乙型肝炎病毒基本核心启动子及前C区突变对疾病进展的影响

目的　研究乙型肝炎病毒 (HBV)基本核心启动子 (BCP)T176 2 /A176 4双突变及前C区A1896突变与肝脏损伤程度的相关性。方法　 2 0 0 0～ 2 0 0 2年哈尔滨医科大学附属第二医院及广东省廉江市医院随机选取 113例慢性HBV感染者 ,采用INNO -LiPA法测定HBVBCPT176 2 /A176 4双突变及前C区A1896突变 ,同时测定HBVS基因序列明确基因型。结果　CHB组、LC组、HCC组BCPT176 2 /A176 4双突变率明显高于AsC组 (分别为 2 4 . 1%比 2. 8% ,χ2 =5 .93,P <0 . 0 5 ;71. 4 %比 2 .8% ,χ2 =2 3 .83,P <0 .0 1和 5 5 . 6 %比 2 .8% ,χ2 =13. 0 9,P <0 . 0 1) ,LC组BCPT176 2 /A176 4双突变率显著高于CHB组 (71 .4 %比 2 4 . 1% ,χ2 =9 .12 ,P <0 .0 1) ;单一C基因型感染者CHB、LC和HCC组BCPT176 2 /A176 4双突变率明显高于AsC组 (分别为 33 .3%比 5 . 3% ,χ2 =3 .89,P <0 .0 5 ;6 9 .2 %比5 . 3% ,P <0 . 0 1和 5 0 .0 %比 5. 3% ,P <0 . 0 5 )。各组前C区A1896突变率均较低 ,在CHB组和LC组无一例发生 ;HCC组前C区A1896突变率与AsC组比较差异无显著性 (11 .1%比 8. 3% ,χ2 =0 . 0 0 ,P >0 . 0 5 )。结论　在慢性HBV感染者中 ,BCPT176 2 /A176 4双突变与慢性肝病进展有关。     

乙型肝炎病毒基因型对乙型肝炎病毒变异的影响

目的探讨乙型肝炎病毒(HBV)基因型对病毒前核心区(前C区,nt1896)及基本核心启动子(BCP,nt1762/1764)变异的影响.方法416例血清HBsAg阳性、HBV DNA定量大于1.0×104拷贝/ml的患者,采用微流基因芯片检测HBV基因型、前C区及BCP变异.结果416例HBV感染者中406例有基因分型结果:B型20.9%、C型65.9%、BC混合型10.8%,10例患者未分出基因型.302例为HBeAg(-)且HBV DNA( )患者,其中248例(82.12%)有前C区或BCP变异,41.06%为前C区变异,31.12?P变异,2种同时变异为9.94%.B型患者前C区变异率为22.9%(20/87),与C型患者前C区变异率39.4%(108/274)及B、C混合型变异率40.0%相比均有显著差异(P＜0.01).在BCP变异及双变异,C型患者变异率均大于B型患者,但差异无统计学意义(P＞0.05).B、C混合型患者前C区及BCP变异率与C型相似.结论HBV基因型可影响病毒前C区及BCP变异,以C型为著.     

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。     

乙型肝炎病毒前C基因区变异与乙肝病毒复制关系的临床研究

目的探讨乙型肝炎病毒（Hepatitis B Virus,HBV）前C基因区变异与HBV-DNA载量的关系。方法通过DNA扩增、基因序列分析检测21例慢性肝炎、18例肝硬化和15例肝癌血清的HBV前C区和基本核心启动子（Basic Core Promoter,BCP）基因序列,荧光定量聚合酶链反应技术定量检测血清中的HBV-DNA。结果野生株与前C区终止变异、BCP双变异以及联合变异组HBV-DNA载量测定差异无显著性（P〉0.05）;BCP双变异HBV-DNA载量HBeAg（-）组显著高于HBeAg（＋）组（P〉0.05）。结论前C区终止变异和BCP双变异对HBV DNA复制无明显影响。HBeAg（-）的慢性肝病患者BCP变异后HBV DNA复制明显活跃。     

孕妇中乙型肝炎病毒前C区和核心启动子区热点变异研究

目的研究乙型肝炎病毒（HBV）携带产妇体内HBV前C区和核心启动子（CP）区热点变异情况。方法82例无症状慢性HBV携带临产孕妇采用HBV核酸扩增荧光定量检测试剂盒抽提孕妇血清HBVDNA,血清HBsAg、HBeAg采用EIA法检测,半巢式PCR法扩增产妇HBV前C区、CP区核苷酸片段,ABI3730型DNA自动荧光测序仪对PCR产物直接测序。结果82例无症状HBV携带临产孕妇中HBsAg单阳性52例,HBsAg、HBeAg双阳性30例;单阳性孕妇中1896G→A变异的检出率为21．2％（11／52）,仅测到1例双阳性孕妇存在1896G→A变异,1899G→A变异仅发生在1例单阳性孕妇,且与1896G→A变异连锁出现。1762A→／1764G→A变异总是连锁出现,且仅发生于单阳性孕妇,单阳性孕妇中双变异的检出率为17．3％（9／52）。结论HBV前C区、CP区1896G→A及1762A→T／1764G→A热点变异在无症状HBV携带单阳性孕妇中有较高的检出率,HBV前C区和核心启动子区的热点变异可能与肝损伤的严重程度无关。     

Prevalence of hepatitis B virus precore stop codon mutations in chronically infected children

AIM: To find out whether there is a significant difference in the prevalence of the precore stop codon mutation between HBeAg positive and anti-HBe positive children. METHODS: We investigated a large pediatric population of 155 European children (mean age 10.9 years) with chronic hepatitis B by PCR and direct sequencing. Ninety were HBeAg positive and 65 had seroconversion to anti-HBe. Additionally genotyping was performed. RESULTS: Seventy-four (48%) of the sequenced HBV strains were attributed to genotype D and 81 (52%) to genotype A. In the group of 90 HBeAg positive patients, 2 (2.2%) 1896-G-to-A transitions leading to precore stop codon mutation were found, and in the group of 65 anti-HBe positive children, 5 (7.7%) were identified harbouring HBeAg-minus mutants. The difference was not statistically significant (P=0.13). CONCLUSIONS: HBeAg minus variants as predominant viral HB strains play a minor role in the course of chronic hepatitis B in European children.     

慢性乙型肝炎患者乙型肝炎病毒前C区基因突变与临床关系

为了解慢性乙型肝炎患者是否有前Ｃ区基因突变及其与临床的关系，应用快速检测乙型肝炎病毒前Ｃ区基因突变方法，对５３例患者进行了研究。结果显示：２５例ＨＢＶＤＮＡ阳性患者中１３例有突变株感染，最常见的突变形式为１８９８位或伴有１９０１位突变（１０／１３例）。     

乙型肝炎病毒前C区A83变异株的临床检测及分析

目的 了解ＨＢＶ前Ｃ区Ａ８３变异的临床意义。方法 用错配ＰＣＲ－ＲＦＬＰ检测ＨＢＶ感染者血清ＨＢＶ前Ｃ区Ａ８３变异株。结果 该变异株在急性肝炎未检出;在慢性肝炎、肝硬化和重型肝炎检出率分别为６６．７％、４８．９％和４５．５％;慢性无症状ＨＢＶ携带者为３．３％;各型肝炎均以血清抗ＨＢｅ（＋）组的变异检出率最高;肝硬化血清变异株阳性者的腹水发生率和肝功功能异常率较阴性者无显著差别。结论 机体对ＨＢｅＡ     

Hepatitis B virus subgenotypes and basal core promoter mutations in Indonesia

AIM： To identify the distribution of hepatitis B virus （HBV） subgenotype and basal core promoter （BCP） mutations among patients with HBV-associated liver disease in Indonesia.
METHODS： Patients with chronic hepatitis （CH, n =61）, liver cirrhosis （LC, n = 62）, and hepatocellular carcinoma （HCC, n = 48） were included in this study. HBV subgenotype was identified based on S or preS gene sequence, and mutations in the HBx gene including the overlapping BCP region were examined by direct sequencing.
RESULTS： HBV genotype B （subgenotypes B2, B3, B4, 85 and B7） the major genotype in the samples, accounted for 75.4%, 71.0% and 75.0% of CH, LC and HCC patients, respectively, while the genotype C （subgenotypes C1, C2 and C3） was detected in 24.6%, 29.0%, and 25.0% of CH, LC, and HCC patients, respectively. Subgenotypes B3 （84.9%） and C1 （82.2%） were the main subgenotype in HBV genotype B and C, respectively. Serotype adw2 （84.9%） and adrq＋ （89.4%） were the most prevalent in HBV genotype B and C, respectively. Double mutation （A1762T/G1764A） in the BCP was significantly higher in LC （59.7%） and HCC （54.2%） than in CH （19.7%）, suggesting that this mutation was associated with severity of liver disease. The T1753V was also higher in LC （46.8%）, but lower in HCC （22.9%） and CH （18.0%）, suggesting that this mutation may be an indicator of cirrhosis.
CONCLUSION： HBV genotype B/B3 and C/C1 are the major genotypes in Indonesia. Mutations in BCP, such as A1762T/G1764A and T1753V, might have an association with manifestations of liver disease.     

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.     

Mutations of polymerase,precore and core promoter gene in hepatitis B virus during 5-year lamivudine therapy

BACKGROUND/AIMS: The effects of long-term lamivudine therapy on changes in polymerase and precore/core promoter mutations are unknown. The aim of this study was to determine the changes in these regions in patients with chronic hepatitis B virus (HBV) infection treated with lamivudine for 5 years. METHODS: Serum samples obtained from 16 patients at the beginning of and during therapy were polymerase chain reaction-amplified, and nucleotide sequences of HBV analyzed. RESULTS: By the end of 5-year therapy, mutations in YMDD motif emerged in ten patients. Mutations in L526M, M550V and M550I in polymerase were found in seven, six and six patients, respectively. The L526M mutant was found at the time or after detection of M550V/I mutant in six of seven patients. At baseline, precore and core promoter mutations were found in eight and 12 of 16 patients, respectively. Mutants of precore and core promoter were replaced by wild-type virus in each of three patients infected with mutants at 1 year. However, at 5 years of treatment, precore and core promoter mutations reappeared in some patients. CONCLUSIONS: Our data showed that lamivudine initially selected from precore/core promoter mutants to wild-type virus, but precore mutation reappeared during prolonged therapy.     

Precore/basal core promoter mutants quantification throughout phases of hepatitis B virus infection by Simpleprobe

AIM: To investigate precore/basal core promoter (PC/BCP) mutants throughout hepatitis B virus (HBV) infection and to determine their relationship to hepatitis B early antigen (HBeAg) titers.METHODS: We enrolled 191 patients in various stages of HBV infection at the Huashan Hospital and the Taizhou Municipal Hospital from 2010 to 2012. None of the patients received antiviral therapy. HBV DNA from serum, was quantified by real-time PCR. The HBV genotype was determined by direct sequencing of the S gene. We used the Simpleprobe ultrasensitive quantitative method to detect PC/BCP mutants in each patient. We compared the strain number, percentage, and the changes in PC/BCP mutants in different phases, and analyzed the relationship between PC/BCP mutants and HBeAg by multiple linear regression and logistic regression.RESULTS: Patients with HBV infection (n = 191) were assigned to groups by phase: Immune tolerance (IT) = 55, Immune clearance (IC) = 67, Low-replicative (LR) = 49, and HBeAg-negative hepatitis (ENH) = 20. Of the patients (male, 112; female, 79) enrolled, 122 were HBeAg-positive and 69 were HBeAg-negative. The median age was 33 years (range: 18-78 years). PC and BCP mutation detection rates were 84.82% (162/191) and 96.86% (185/191), respectively. In five HBeAg-negative cases, we detected double mutation G1896A/G1899A. The logarithm value of PC mutant quantities (log₁₀ PC) significantly differed in IT, IC, and LR phases, as well as in the ENH phase (F = 49.350, P < 0.001). The logarithm value of BCP mutant quantities (log₁₀ BCP) also differed during the four phases (F = 25.530, P < 0.001). Log₁₀ PC and log₁₀ BCP values were high in the IT and IC phases, decreased in the LR phase, and increased in the ENH phase, although the absolute value at this point remained lower than that in the IT and IC phases. PC mutant quantity per total viral load (PC%) and BCP mutant quantity per total viral load (BCP%) differed between phases (F = 20.040, P < 0.001; F = 10.830, P < 0.001), with PC% and BCP% gradually increasing in successive phases. HBeAg titers negatively correlated with PC% (Spearman’s rho = -0.354, P < 0.001) and BCP% (Spearman’s rho = -0.395, P < 0.001). The negative correlation between PC% and HBeAg status was significant (B = -5.281, P = 0.001), but there was no such correlation between BCP% and HBeAg status (B = -0.523, P = 0.552).CONCLUSION: PC/BCP mutants become predominant in a dynamic and continuous process. Log₁₀ PC, log₁₀ BCP, PC% and BCP% might be combined to evaluate disease progression. PC% determines HBeAg status.     

慢性HBV感染者抗病毒治疗前HBV基本核心启动子及前C区突变的检测及意义

目的研究慢性乙型肝炎病毒（HBV）感染者抗病毒治疗前HBV基本核心启动子（BCP）突变和前C区（PreC）突变与HBeAg、HBV DNA水平和慢性肝病进展的关系。方法收集283例慢性HBV感染者抗病毒治疗前的血清标本,其中慢性乙型肝炎（CHB）185例,肝硬化（LC）98例。采用PCR后直接测序法检测HBV BCP和PreC区突变,同时确定基因型。结果在HBeAg阴性和HBeAg阳性CHB患者中,前C区A1896变异率分别为44.6%（37/83）和21.6%（22/102）（χ2=11.154,P=0.001）,LC患者分别为43.4%（23/53）和17.0%（8/47）（χ2=8.101,P=0.004）。在HBeAg阳性患者中,BCP T1762/A1764双突变率LC组和CHB组分别为89.4%（42/47）和70.6%（72/102）（χ2=6.310,P=0.012）。在单变量分析中,只有年龄（≥45岁）（χ2=27.861,P〈0.001）、BCP T1762/A1764双突变（χ2=8.675,P=0.003）和HBV DNA（≥105拷贝/ml）（χ2=20.499,P〈0.001）与LC进展有关。多因素Logistic回归分析（匹配年龄和性别）发现,BCP T1762/A1764双突变（OR=3.260,95%CI：1.401～7.586;wald=7.517,P=0.006）和HBV DNA（≥105拷贝/ml）（OR=4.640,95%CI：2.331～9.237;wald=19.089,P〈0.001）是LC进展的危险因素。结论前C区A1896突变与HBeAg的消失有关;年龄（≥45岁）、BCP T1762/A1764双突变和HBV DNA高载量（≥105拷贝/ml）与肝硬化进展有关。     

乙型肝炎病毒前C/BCP区变异的研究现状

HBV的前C区及基本核心启动子（BCP）变异可降低HBeAg的合成与分泌,也可增加病毒复制能力,导致HBeAg阴性慢性乙型肝炎（CHB）,对临床产生重要的影响。本文就HBV前C/BCP区变异的分子机理、生物学意义、实验室检测以及对临床和抗病毒治疗的影响等方面的研究进展作一综述。     

乙型肝炎病毒C基因启动子和前C基因变异与HBeAg含量和肝炎病情的临床研究

研究乙型肝炎病毒(HBV)C基因启动子(CP)和前C基因变异对HBeAg表达和病情的影响。通过DNA扩增、基因序列分析检测48例慢性乙肝和12例慢性重型乙肝患者血清的HBV CP和前C基因序列,及通过微粒子发光法定量检测血清中HBeAg的含量。(1)前C终止变异(nt1896G→A)在重型乙型肝炎病例中的发生率显著升高(66.7％);CP双变异(nt1762A→T和1764G→A)则在慢性乙型肝炎中度和重度的病例中的发生率显著升高(分别为52.6％和54.5％)。(2)双变异组和终止变异组的HBeAg含量均显著下降,P<0.01。但终止变异组HBeAg含量的下降较双变异组更为明显,P<0.05,且eAb阳性率也显著升高,P相似文献   

Association of core promoter mutations of hepatitis B virus and viral load is different in HBeAg（＋） and HBeAg（-） patients

AIM:To identify the prevalence of hepatitis B e antigen (HBeAg) and to assess the association of hepatitis B virus (HBV) core promoter mutations and viral load in Indonesian patients.METHODS:Sixty-four patients with chronic hepatitis,65 with liver cirrhosis and 50 with hepatocellular carcinoma were included in this study.HBeAg and hepatitis B e antibody (HBeAb) tests were performed using enzyme-linked immunosorbent assay and the mutations were analyzed by sequencing.Viral load was measured by real-time poly...     

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.