Nucleic acid sequence analysis of basal core promoter/precore/core region of hepatitis B virus isolated from chronic carriers of the virus from Kolkata, eastern India: low frequency of mutation in the precore region

OBJECTIVE: The aim of the present study was to characterize the predominant hepatitis B virus (HBV) strains and their molecular variants present in the HBV isolates of the different genotypes found among the chronic carriers of the virus in our community. METHODS: Precore/core and core promoter regions of HBV DNA were amplified by polymerase chain reaction and then subjected to direct sequencing. Of the 64 hepatitis B surface antigen (HBsAg)-positive chronic HBV carriers investigated, 44 were HBeAg negative and 20 were HBeAg positive. RESULTS: In addition to genotype D, which was the predominant genotype, 12 genotype C (18.7%) and 6 genotype A (9.4%) were also detected. Presence of T at nt 1858 has often been related to the development of precore stop mutation at nt 1896, while that of C has been related to the development of 1762-1764 double mutation. In our study group, 39 of the 44 HBeAg-negative samples have T1858. The precore stop codon mutation was found in only 8 (18%) of the HBeAg-negative samples. More than half of the HBeAg-negative samples had wild-type sequence in the precore region. The core promoter region could be sequenced from 40 samples, and 1762-1764 double mutation was detected in 13 (32.5%) of them. No significant changes could be detected in the core amino acid sequence of these isolates. CONCLUSION: The pattern of core promoter and precore mutation of HBV isolates in the present study is atypical and not in accordance with reports from other parts of the world, where genotype D and genotype C with T at codon 1858 are common.     

Should treatment of hepatitis B depend on hepatitis B virus genotypes? A hypothesis generated from an explorative analysis of published evidence

OBJECTIVE: Standard treatment for hepatitis B is either based on interferon or on nucleos(t)ide analogues. Available evidence on treatment outcome by hepatitis B virus genotype was summarized and analysed to determine whether the genotype has a substantial role in selecting treatment. METHODS: Abstracts from PubMed, the European Association for the Study of the Liver and the American Association for the Study of the Liver Diseases were screened for publications with at least 30 patients and reporting hepatitis B treatment results (hepatitis B e antigen [HBeAg] seroconversion, loss of HBeAg or loss of HBV DNA) broken down by hepatitis B genotypes. Twenty reports were identified and included. Forest-plot techniques were applied to visualize the association of HBV genotypes (A versus D and B versus C) with treatment outcome given by type of treatment and HBeAg status. The potential size of a treatment by genotype interaction was estimated. RESULTS: Treatment response to nucleos(t)ide analogues is not significantly influenced by HBV genotype in HBeAg-positive or HBeAg-negative individuals. In contrast, HBV genotypes are informative concerning responses to interferon treatment in all patients with genotype A versus D and in HBeAg-positive patients with genotype B versus C. CONCLUSION: If no contraindications are present, interferon may be considered as first-line therapy in all genotype A patients and in individuals with genotype B who are HBeAg positive. However, confirmation of this observation in a prospective clinical trial is warranted, because this analysis is explorative and hypothesis generating only.     

520例慢性乙型肝炎HBeAg阴性与阳性患者临床和HBV-DNA比较分析

目的:了解HBeAg(-)慢性乙型肝炎(CHB)患者临床特点和血清HBV-DNA检测的意义。方法:对520例CHB患者的住院病历进行回顾性调查,分析HBeAg(-)和HBeAg( )CHB患者HBV-DNA定量组内和组间差异。结果:HBeAg(-)CHB 166例,占31.92%;HBeAg( )CHB 354例,占68.08%。HBeAg( )组HBV-DNA总体上高于HBeAg(-)组;不同病程HBeAg(-)CHB比例轻度组为12.8%,中度组为13.2%,重度组为19.2%,重型组为31.4%,肝硬化组为47.5%。结论:目前我国CHB病例以HBeAg( )者占多数,HBeAg( )组HBV-DNA总体上高于HBeAg(-)组,随着病程进展HBeAg(-)CHB比例上升。     

Sequential accumulation of the basal core promoter and the precore mutations in the progression of hepatitis B virus-related chronic liver disease

OBJECTIVES: Despite the pathogenic role of the basal core promoter (BCP) and the precore mutations in chronic hepatitis B virus (HBV) infection, their role in the progression of liver disease is still controversial. We analyzed whether the accumulation of these mutations might enhance the progression of HBV-related chronic liver disease. METHODS: Forty consecutive patients at each clinical status were analyzed. Clinical statuses were as follows: HBeAg-positive asymptomatic carrier (HBeAg(+) ASC) (defined as HBeAg(+), anti-HBe(-), HBV-DNA(+) by hybridization, normal ALT); inactive HBsAg carrier; chronic hepatitis B; liver cirrhosis. The genotype and the BCP/precore regions were determined by PCR using genotype specific primers and direct sequencing, respectively. RESULTS: All patients except one were infected with genotype C. The A to T mutation at nucleotide 1762 and/or G to A mutation at nucleotide 1764 were found in 30% in HBeAg(+) ASC, 65.7% in inactive HBsAg carrier, 95% in chronic hepatitis B, and 90% in liver cirrhosis (p < 0.001). The prevalence of the G to A mutation at nucleotide 1896 was 5% in HBeAg(+) ASC, 22.5% in inactive HBsAg carrier, 32.5% in chronic hepatitis B, and 50% in liver cirrhosis, respectively (p < 0.001). The T to C/A mutation at nucleotide 1753 in the BCP and G to A mutation at nucleotide 1899 in the precore were more frequent in liver cirrhosis than in the other clinical statuses (p < 0.05). CONCLUSION: Sequential accumulation of mutations in the BCP/precore has an important role in the progression of HBV-related liver disease.     

Precore wild-type hepatitis B virus with G1896 in the resolution of persistent hepatitis B virus infection

OBJECTIVE: Factors influencing the resolution of persistent hepatitis B virus (HBV) infection were sought for. METHODS: The loss of hepatitis B surface antigen (HBsAg) from serum was correlated with mutations in HBV DNA for a hepatitis B e antigen (HBeAg)-minus phenotype in patients infected with HBV genotype C and positive for HBeAg at presentation. RESULTS: HBeAg turned negative in all the 22 patients in whom HBV infection resolved, but only in 11 of the 25 patients with severe liver diseases (100 vs. 44%, p = 0.0001). The precore wild type (G1896) persisted significantly more frequently in the 22 patients in whom HBV infection resolved than in the 11 patients who developed decompensated liver cirrhosis or hepatocellular carcinoma (15/22 or 68% vs. 1/11 or 9%, p = 0.005). The double mutation in the core promoter (T1762/A1764) was comparably frequent in the two groups of patients at presentation (14/22 or 64% vs. 7/11 or 64%) and >15 years thereafter (18/22 or 82% vs. 10/11 or 91%). CONCLUSION: The precore wild type (G1896) would seem to facilitate the resolution of HBV infection, while the precore mutant (A1896) may induce severe liver diseases in patients with HBeAg-positive chronic hepatitis who have lost HBeAg from serum.     

血清HBsAg和HBV DNA对慢性乙型肝炎肝组织病理状态的判别评价

目的构建基于血清HBsAg水平、HBV DNA载量、HBsAg/HBV DNA比值判别慢性乙型肝炎肝组织不同病理学分级和分期的Fisher判别函数,评价Fisher判别函数判别肝组织不同病理学分级和分期的效能。方法 472例经肝组织活检的慢性乙型肝炎患者入选本研究,其中HBeAg阳性279例,HBeAg阴性193例。血清HBsAg采用Abbott Architect I2000及其配套试剂检测,血清HBV DNA采用实时荧光定量PCR检测。统计分析采用SPSS 13.0软件。结果 HBeAg阳性患者血清HBsAg、HBV DNA与病理学分级和分期呈显著负相关（P〈0.05）;血清HBsAg/HBV DNA与病理学分级呈显著负相关（P〈0.01）,与病理学分期无显著相关性（P〉0.05）。HBeAg阴性患者血清HBsAg与病理学分级和分期均无显著相关性（P〉0.05）,血清HBV DNA、HBsAg/HBV DNA分别与病理学分级和分期呈显著正相关和负相关（P〈0.01）。根据Bayes逐步判别分析,HBeAg阳性和阴性患者符合判别不同病理学分级的模型纳入自变量标准的指标分别只有血清HBsAg和HBsAg/HBV DNA,符合判别不同病理学分期的模型纳入自变量标准的指标分别有血清HBsAg、HBV DNA和HBV DNA。判别HBeAg阳性患者不同病理学分级的Fisher判别函数判别G1、G2、G3的一致率分别为5.8%、51.1%、59.1%,判别HBeAg阴性患者不同病理学分级的Fisher判别函数判别G1、G2、G3的一致率分别为95.5%、0.0%、5.7%;判别HBeAg阳性患者不同病理学分期的Fisher判别函数判别S1、S2、S3、S4的一致率分别为36.4%、34.9%、21.6%、57.9%,判别HBeAg阴性患者不同病理学分期的Fisher判别函数判别S1、S2、S3、S4的一致率分别为86.7%、29.4%、0.0%、0.0%。结论基于血清HBsAg和基于血清HBsAg、HBV DNA的Fisher判别函数分别对HBeAg阳性患者肝组织病理学分级G3和分期S4有重要的判别价值;基于HBsAg/HBV DNA比值和基于血清HBV DNA的Fisher判别函数对HBeAg阴性患者肝组织病理学分级G1和分期S1有重要的判别价值。     

Treatment of patients with chronic hepatitis B who have failed previous antiviral treatment with pegylated interferon alpha2a (40 kda; PEGASYS)

BACKGROUND: Although nucleot(s)ide analogues can effectively suppress hepatitis B virus (HBV) replication, many patients experience relapse of hepatitis after cessation of treatment. We aimed to investigate the efficacy of pegylated interferon alpha2a (PEG-IFN-alpha2a) in these difficult-to-treat patients. METHODS: Chronic hepatitis B patients who have received antiviral drugs for > or =12 months and stopped for > or =6 months were treated by 48-week PEG-IFN-alpha2a. Virological response was defined as HBV DNA <10,000 copies/ml and hepatitis B e antigen (HBeAg) seroconversion (for HBeAg-positive patients). RESULTS: A total of 40 patients, 29 HBeAg-positive and 11 HBeAg-negative, with median log10 HBV DNA 7.3 copies/ml and alanine aminotransferase 110 IU/ml were studied. The last antiviral treatment was given for 92 +/- 61 weeks and stopped for 176 +/- 88 weeks. At the end of treatment, 22 (12 HBeAg-positive and 10 HBeAg-negative; 55%) patients had virological response and 16 (7 HBeAg-positive and 9 HBeAg-negative; 40%) patients had undetectable HBV DNA (<100 copies/ml). At 24 weeks post-treatment, 14 (8 HBeAg-positive and 6 HBeAg-negative; 35%) patients had virological response and 9 (5 HBeAg-positive and 4 HBeAg-negative; 23%) patients had undetectable HBV DNA. Two (5%) patients had lost hepatitis B surface antigen. HBV DNA levels at week 24 best predicted sustained virological response (area under curve 0.76, 95% confidence interval 0.60-0.92, P=0.007). At HBV DNA cutoffs of 3 logs and 5 logs at week 24, the sensitivity/specificity for sustained virological response were 50%/85% and 86%/62%, respectively. CONCLUSIONS: PEG-IFN-alpha2a was effective in the treatment of chronic hepatitis B patients who have failed previous antiviral treatment.     

肝癌患者血清中乙型肝炎病毒前C和BCP区基因变异的临床研究

目的了解常州地区乙型肝炎病毒(HBV)DNA前C区(1896)、BCP区(1762/1764)基因的变异,探讨与肝癌的相关性。方法运用基因芯片和核苷酸序列分析技术,对HBVDNA进行分析。结果前C区1896位、BCP区1762/1764位突变普遍存在。(1)在39例HBeAg( )标本组中1896突变为5例(12.8%),1762/1764突变为15例(38.5%);在75例HBeAg(-)组中1896突变为26例(34.7%),1762/1764突变为55例(73.3%)。(2)在114例标本中,慢性乙型肝炎、慢性重型乙型肝炎、乙肝肝硬化、肝癌组中前C区1896、BCP区1762/1764位的总突变率分别为55.9%、71.4%、75.7%、88.9%。结论前C区1896、BCP区1762/1764位突变与HBeAg(-)显著相关,两者总突变率在肝癌患者中显著升高,尤其前C1896、BCP区1762/1764同时突变与肝硬化和肝癌密切相关。     

Classifying hepatitis B virus genotypes

In 1988, hepatitis B virus (HBV) was classified into four genotypes by a sequence divergence in the entire genome exceeding 8%, and designated by capital letters of the alphabet from A to D. There are seven genotypes of HBV (A-G) at present, and an eighth is on the horizon. They have an uneven geographical distribution, and only a few of them are prevalent in a given area of the world. Thus genotype A is frequent in northwest Europe, Sub-Saharan Africa, India and the North, Central and South America, B as well as C are common in Southeast Asia and Oceania, and D is prevalent in the Mediterranean area, Central Asia and South America. Genotype E is restricted to West Africa, and F is localized in Central and South America. The distribution of genotype G added to the alphabet list very recently has yet to be determined. Coinfection with HBV of distinct genotypes is not infrequent and found in about 10% of infected individuals, and is responsible for intertypic recombination of HBV genomes. The mutation for a stop codon in the precore region (G1896A) for aborting the translation of hepatitis B e antigen (HBeAg) is prohibited in HBV genomes of genotype A, as well as some of genotypes C and F, because they possess C at position 1858 that makes a Watson-Crick pair with G at position 1896. Hence, seroconversion to antibody to HBeAg is forbidden or delayed in individuals who carry them. Evidence is accumulating as regards the influence of HBV genotypes on the progression of chronic hepatitis B and response to antiviral therapies. HBV isolates even of the same genotype can differ in virological and clinical characteristics, and therefore, the genotype needs to be classified further into subtypes, especially if they are clinically relevant.     

肝硬化病原学分析和乙肝肝硬化中HBeAg阳性与阴性患者的临床特点比较

目的 通过大样本横断面回顾性调查，了解临床肝硬化患者病原组成及临床所见HBeAg(－)和HBeAg(＋)两类肝硬化患者临床相关因素的异同。 方法 对599例肝硬化患者的住院病历进行回顾性调查，分析肝硬化患者病因构成及占主要病因的乙肝肝硬化中的HBeAg(－)和HBeAg(＋)肝硬化组年龄、ALT、HBV DNA定量、CTP评分、PLT等指标的组内和组间差异。 结果 在肝硬化病因中以病毒性肝炎后肝硬化常见，有557例，占93.0％，其中又以乙肝肝硬化居多。在乙肝肝硬化中HBeAg (－)乙肝肝硬化306例，占62.3％；HBeAg(＋)肝硬化185例，占37.7％。HBeAg(＋)组ALT、HBV DNA总体上均高于HBeAg(－)组。HBeAg(－)组患者年龄明显高于HBeAg(＋)组。两组之间Child-Pugh，（CTP）积分无明显差异。 结论 目前我国临床所见肝硬化病例以病毒性肝炎感染后所致占绝大多数，乙肝肝硬化中又以HBeAg(－)者占多数。HBeAg(－) 与HBeAg(＋)肝硬化患者在病情轻重方面无明显差异，无论HBeAg是否阳性，病毒载量高时，抗病毒治疗及控制肝脏炎症能减轻病情恶化。     

Markers of disease activity in chronic hepatitis B virus infection

BACKGROUND: Assessment of disease activity is important in the management of chronic hepatitis B virus (HBV) infection. Our objective was to study the correlation between serum HBV DNA levels and HBV e antigen (HBeAg) status, alanine aminotransferase (ALT) levels, histologic activity, age and sex in patients who had chronic HBV, with emphasis on those who were HBeAg negative with high replication but had normal or below-normal liver enzyme levels and mild liver disease. METHOD: At our university-affiliated tertiary care medical centre in Turkey, we studied prospectively 179 consecutive patients who were long-term hepatitis B surface antigen carriers. These patients were first separated into 2 groups according to HBeAg positivity and then subdivided into 4 groups according to the presence of HBV DNA, HBeAg status and ALT levels. The clinical, virologic and histologic differences in these patients were evaluated with respect to the HBeAg status. RESULTS: Of the 179 patients, 120 (67%) were HBeAg positive and 59 (33%) were HBeAg negative. The mean (and standard deviation) age in the former group was 24.8 (7.60) and in the latter group was 32.2 (11.2) years (p < 0.001). HBeAg-negative patients had significantly more severe liver disease, more male predominance and lower serum HBV DNA levels than HBeAg-positive patients (p < 0.05). HBeAg status had a close correlation with age. There was a significant correlation between age and serum HBV DNA levels but not between HBV DNA levels and disease activity in study groups. We found that some of anti-HBe-positive patients had below-normal ALT levels with minimal or absent histologic changes despite high viral replication. CONCLUSIONS: Monitoring of ALT is of value in assessing hepatocellular damage in patients with chronic hepatitis B virus infection. HBeAg-negative patients with elevated ALT levels and some with normal ALT levels should be considered highly infectious in the course of chronic HBV infection.     

Natural history of hepatitis B virus infection: an update for clinicians

Hepatitis B virus (HBV) is a common viral pathogen that causes a substantial health burden worldwide. Significant progress has been made in the past few decades in understanding the natural history of HBV infection. A dynamic balance between viral replication and host immune response is pivotal to the pathogenesis of liver disease. In immunocompetent adults, most HBV infections spontaneously resolve, whereas in most neonates and infants they become chronic. Those with chronic HBV may present in 1 of 4 phases of infection: (1) in a state of immune tolerance, (2) with hepatitis B e antigen (HBeAg)positive chronic hepatitis, (3) as an inactive hepatitis B surface antigen carrier, or (4) with HBeAg-negative chronic hepatitis. Of these, HBeAg-positive and HBeAg-negative chronic hepatitis may progress to cirrhosis and its long-term sequelae including hepatic decompensation and hepatocellular carcinoma. Several prognostic factors, such as serum HBV DNA concentrations, HBeAg status, serum aminotransferases, and certain HBV genotypes, have been identified to predict long-term outcome. These data emphasize the importance of monitoring all patients with chronic HBV infection to identify candidates for and select optimal timing of antiviral treatment, to recognize those at risk of complications, and to implement surveillance for early detection of hepatocellular carcinoma.     

乙型肝炎e抗原阴、阳性慢性乙型肝炎患者细胞免疫状态与乙型肝炎病毒DNA水平相关分析

目的 探讨乙型肝炎e抗原(HBeAg)阴性及HBeAg阳性慢性乙型肝炎患者在细胞免疫状态及乙型肝炎病毒DNA(HBV DNA)水平之间的相关性.方法 分别应用流式细胞仪、荧光定量聚合酶链反应(PCR)方法对HBeAg阴、阳性慢性乙型肝炎患者T淋巴细胞亚群及HBV DNA进行检测.结果 HBeAg阴性慢性乙型肝炎患者的HBV DNA水平与CD8 T淋巴细胞比例之间存在正相关,而与CD4/CD8比值之间存在负相关,γ值分别为0.567,-0.601,均P＜0.01;与总T、CD4 T淋巴细胞比例无相关关系.HBeAg阳性组患者的HBV DNA水平与总T、CD4 T、CD8 T淋巴细胞水平相关关系.结论 HBeAg阴性慢性乙型肝炎患者HBV DNA水平与机体细胞免疫密切相关.     

Low HBeAg serum levels correlate with the presence of the double A1762T/G1764A core promoter mutation and a positive response to interferon in patients with chronic hepatitis B virus infection

OBJECTIVE: To investigate the correlation of serum hepatitis B e antigen (HBeAg) levels with the presence of core promoter (CP) mutations, hepatitis B virus (HBV) viremia and the response to interferon (IFN) in patients with chronic hepatitis B. METHODS: Fourteen HBeAg-positive patients received alpha-2a IFN. Diluted serum samples of responders were tested for HBeAg positivity at dilutions of 1:40, 1:160 and 1:640 at the following time points: T0 (before starting IFN), T1 [at peak alanine aminotransferase (ALT) preceding HBeAg seroconversion], T2 (at ALT normalisation) and T3 (end of treatment). Nonresponder samples were similarly tested at times T0 and T3. The HBV CP and precore regions were sequenced at the same time points as for HBeAg testing. RESULTS: Six of 14 patients (43%) responded to IFN treatment and had lower HBeAg levels than nonresponders at T0 (p = 0.003). Five of 6 responders (83%) and none of the nonresponders had the A1762T/G1764A CP mutations (0/8, p < 0.003). At T0, HBeAg was negative at the 1:640 dilution in 5 of the 6 responders, who also had lower HBV DNA levels than nonresponders (p = 0.003). During IFN treatment, HBeAg levels decreased and HBV DNA became negative at T1 in responders. CONCLUSIONS: Low serum HBeAg and HBV DNA levels correlate with the presence of CP mutations and response to IFN treatment and can be considered as predictive markers of response to IFN.     

乙型肝炎病毒基因型与YMDD基因区序列突变及BCP突变关系探讨

目的 对湘潭地区HBV感染者的基因型、YMDD基因区序列突变及BCP区突变情况进行研究,并对三者之间的关系进行探讨.方法 针对湘潭地区952例不同类型的HBV感染者的样本同时进行基因型、YMDD基因区序列突变及BCP区突变检测,并对检测结果进行统计分析.结果 湘潭地区HBV各种基因型分布比例为:B型698例、占73.32%,C型115例、占12.08%,B、C型混合感染139例、占14.60%.YMDD基因区序列突变结果显示:YMDD野生型844例、占88.66%,其余为YMDD突变型,其中YVDD 54例、占5.67%,YIDD 53例、占5.57%,1例为YMDD与YVDD混合感染.BCP区突变结果显示:1762A/1764G(野生型)672例、占70.59%,1762T/1764A(突变型)188例、占19.75%,其余92例为1762T/1764A和1762A/1764G混合型、占9.66%.基因型、YMDD基因区序列突变及BCP区突变三者相关性分析显示:HBV B型和C型YMDD基因区序列突变率分别为10.04%、10.43%,差异无统计学意义(χ2=0.017,P＞0.05),HBV B型和C型YMDD基因区序列突变类别差异有统计学意义(χ2=4.836,P＜0.05),HBV C型YVDD突变率(9.57%)要高于B型(5.88%).HBV B型和C型BCP区突变率分别为27.36%、46.09%,差异有统计学意义(χ2=16.478,P＜0.01),C型BCP区突变率要高于B型.HBV YMDD野生型和突变型的BCP区突变率分别为2 8.67%、35.51%,差异无统计学意义(χ2=2.139,P＞0.05),但YVDD BCP区突变率(61.11%)要高于其他类别.结论 (1)湘潭地区流行的HBV基因型主要为B型和C型,其中B型为优势基因型,具有南方地区的特点.(2)拉米夫定治疗前通过HBV基因型检测来预测抗病毒应答可能并无实际意义.(3)HBV基因分型、YMDD基因区序列突变、BCP区突变检测的应用,将有助于临床上对乙肝患者的预后和转归进行正确评价,为及时合理的实施干预措施提供了重要依据.

Abstract：

Objective To investigate the relationship between HBV genotypes and YMDD motif mutations or BCP mutations in Xiangtan of Hunan Province. Methods HBV genotypes, YMDD motif mutations and BCP mutations were analyzed in 952 HBV infected patients. Results HBV genotyping showed that 698 HBV type B patients and 115 HBV type C patients accounted for 73.32% and 12.08% respectively of all the participants. The rest 139( 14.60% )were genotype B and C mixed infection( B + C ). The analysis of YMDD motif mutations showed that 844 YMDD wild-type which accounted for 88.66% of all the subjects and the remainder were YMDD mutation types, of which 54( 5.67% ) carried YVDD, 53( 5.57% ) YIDD,and 1 YVDD and YIDD mixed infection. Basic Core Promoter mutations showed that 1762A/1764G ( wild type )accounted for 70.59% and 1762T/1764A( mutant ) accounted for 19.75%. The rest 92 patients were 1762T/1764A and 1762A/1764G mixed infection. This study showed no significant difference in the rate of YMDD mutation( 10.04% vs 10.43% ,χ2 =0.017,P＞0.05 ) ,but a significant difference in the types of YMDD mutation(χ2 = 4.836, P ＜ 0.05 )between HBV types B and C. The YVDD mutation was more commonly seen in genotype C( 9.57% ) than in genotype B( 5.88% ). The BCP mutation rate showed a significant difference( 27.36% vs 46.09%, χ2 = 16.478, P ＜ 0.01 ). Genotype C was more frequent than genotype B. The BCP mutation rate showed no significant difference between YMDD Wild-type and YMDD mutation types( 28.67% vs 35.51%, χ2 = 2.139, P ＞ 0.05 ), but most of BCP mutations happened in YVDD mutant type( 61.11% ). Conclusions ( 1 ) The predominant HBV genotypes in Xiangtan were genotype B and genotype C, the major genotype was type B, which display the characteristics of epidemiology in Southern China. ( 2 ) Determination of HBV genotypes before lamivudine therapy was probably not an important pretreatment investigation to predict antiviral responses. ( 3 ) Detection of HBV genotypes, YMDD motif mutations and BCP mutations will contribute to the correct evaluation of prognosis and timely proper management of HBV patients.     

乙型肝炎病毒大蛋白在血清HBeAg阴性HBV感染者中的检测意义

目的探讨乙型肝炎病毒大蛋白（HBV-LP）在血清HBeAg阴性HBV感染者中的检测意义。方法对162例HBV感染者血清采用酶联免疫吸附试验检测乙型肝炎病毒大蛋白及乙型肝炎病毒标志物,FQ-PCR定量检测HBV-DNA。结果162例HBV感染者血清中,HBV-LP与HBV-DNA、HBeAg之间均关联显著（P均〈0.01）;HBV-LP与HBV-DNA、HBeAg间阳性率均存在显著性差异（P均〈0.05）;HBV-LP阳性率为84.57%,较HBV-DNA、HBeAg高,分别为62.96%、38.89%（P均〈0.05）;血清HBV-LP浓度及其阳性率在HBeAg和HBV-DNA的阳性组与阴性组间均存在显著性差异（P均〈0.05）;HBeAg阳性组中HBV-LP与HBV-DNA间阳性率无显著性差异（P〉0.05）,检测符合率为96.83%;HBeAg阴性组中HBV-LP与HBV-DNA间阳性率存在显著性差异（P〈0.05）;HBV-LP阳性率为76.77%,较HBV-DNA高。在58例HBeAg和HBV-DNA共同阴性的HBV感染血清中,检出HBV-LP阳性42例。结论血清HBV-LP是监测血清HBeAg阴性HBV感染者体内病毒复制、疾病进程与疗效的敏感指标。     

强效核苷类似物抗病毒过程中血清HBV DNA变化和HBeAg血清学转换分析

目的:探讨恩替卡韦及替比夫定治疗慢性乙型肝炎(慢乙肝)过程中血清HBV DNA水平变化和HBeAg血清学阴转率的差异.方法:选择初治慢乙肝患者126例,按其治疗方案分为恩替卡韦0.5 mg/d治疗组(恩替卡韦组)83例和替比夫定600 mg/d治疗组(替比夫定组)43例,观察两组慢乙肝患者治疗前及治疗4、8、12、24、48周时血清HBV DNA水平、阴转率及其下降模式,比较治疗48周时HBeAg血清学阴转率.结果:治疗4周时思替卡韦组和替比夫定组HBeAg阳性患者HBV DNA水平分别为(5.57±1.30)lg copies/L、(5.76±1.34)lg copies/L,两组HBeAg阴性患者则分别为(5.75±1.01)lg copies/L、(6.03±1.86)lg copies/L,两组HBV DNA水平与治疗前比较差异均有统计学意义(P均<0.05),但两组组间比较差异无统计学意义(P>0.05).两组HBeAg阳性患者各个时间点HBV DNA阴转率比较差异均无统计学意义(P>0.05);恩替卡韦组HBeAg阴性患者除24周外的HBV DNA阴转率均明显高于替比夫定组HBeAg阴性患者(P均<0.05).无论是HBeAg阳性患者还是HBeAg阴性患者,两组的病毒下降模式比较差异无统计学意义(P>0.05).两组治疗48周HBeAg阴转率比较差异无统计学意义(P>0.05).结论:两种核苷类似物均能快速、强效抑制HBV DNA;对于HBeAg阴性患者,恩替卡韦治疗较替比夫定治疗可获得更高的HBV DNA阴转率.     

初诊慢肝患者血清中乙肝病毒DNA载量与血清标志物的关系

目的探讨初诊慢性乙型肝炎血清标志物与HBV DNA的关系。对象与方法采用酶联免疫方法和荧光定量聚合酶链反应分别对512例不同乙肝病人及202例HBeAg阴性、257例HBeAg阳性慢肝病人进行血清标志物、HBV DNA定量检测并分析结果。结果慢肝组有315例(68.9%)病毒颗粒浓度高于107,肝硬化组7例(13.2%)病毒颗粒浓度高于107;HBeAg阳性慢肝组245例(95.3%)患者病毒颗粒含量高浓度,HBeAg阴性慢肝组68例(33.7%)患者病毒颗粒含量在105-7copy/mL区段。结果慢肝组与肝硬化组相比,高浓度病毒以慢肝组中最明显,病情发展至肝硬化期病毒含量多数不高;慢肝病人中不同乙肝血清标志物模式其病毒含量分布特点不同,与HBeAg阳性慢肝组相比HBeAg阴性组体内病毒复制水平低(p<0.01),传染性较前者弱。最常见前C区突变,导致HBeAg阴性CHB病情加重和病毒复制增加。     

Clinical significance of the genotype and core promoter/pre-core mutations in hepatitis B virus carriers

It has been shown that clinical and virological characteristics vary among hepatitis B virus (HBV) genotypes. In this study, we measured the virus level, disease severity, and presence or absence of core promoter (CP)/pre-core (PC) mutations in 241 HBV carriers, and investigated the clinical significance of measuring the HBV genotype. In genotype C HBV carriers, the proportion of hepatitis B e antigen (HBeAg)-positive patients was significantly higher than that in genotype B HBV carriers (0 vs. 34.4%, p < 0.05), and the virus level was higher (4.9 vs. 4.05 LGE/ml). In the genotype B HBV carriers, the incidence of PC mutation was significantly higher (69 vs. 34%, p < 0.05). In the genotype C HBV carriers, the incidence of CP mutation was significantly higher (13 vs. 78%, p < 0.05). We compared patients with the wild (W)/mutant (M) pattern in the CP/PC regions to those with the M/W pattern in the CP/PC regions among the genotype C HBV carriers. Both the proportion of HBeAg-positive patients (65.8 vs. 15.4%, p < 0.05) and the alanine aminotransferase (ALT) level (48 vs. 21.5 IU, p < 0.05) were higher in the patients with the M/W pattern in the CP/PC regions, and the disease severity was deteriorated. In conclusion, genotype B HBV may more frequently induce HBe seroconversion via PC mutation compared to genotype C HBV. Among the genotype C HBV carriers, hepatitis activity and the deterioration of the disease severity were significantly inhibited in the group in which PC mutation initially occurred, in comparison to the group in which CP mutation initially occurred.     

421例乙肝病毒e抗原阴性及阳性慢性乙型肝炎及相关肝病临床和病毒学特点比较分析

目的：了解乙肝病毒e抗原（HBeAg）阴性和阳性慢性乙型肝炎及相关肝病患者临床和病毒学特点。方法：对421例慢性乙型肝炎及相关肝病患者进行回顾性调查,分析HBeAg阴性和阳性的乙肝患者流行病学、病例构成、肝功能、HBV-DNA载量及乙肝前S1抗体（PreS1Ag）等指标的差异。结果：HBeAg阴性组发病率高,占57.96%,高于HBeAg阳性组,其重型肝炎、肝硬化、肝癌构成及肝功能损害指标高于HBeAg阳性组,HBV-DNA载量、PreS1Ag阳性率低于HBeAg阳性组。结论：HBeAg阴性的慢性乙型肝炎及相关肝病发病率高,进展快,预后差,是今后防治慢性乙型肝炎的重点。