蚌埠地区乙型肝炎病毒基因分型及临床意义

目的 了解蚌埠地区乙型肝炎患者的病毒(HBV)基因型分布情况,并探讨其临床意义。方法 选择在我院就诊的乙型肝炎患者268例,采用荧光定量PCR测定乙肝患者的病毒载量并进行基因分型,同时分析基因型与患者年龄及HBeAg的关系。结果 268份血清标本中有202株为C基因型,占75.37%(202/268),46株为B基因型,20株为B+C型;C基因型的病毒载量和HBeAg阳性率明显高于其他基因型,B基因型感染者的年龄相对较轻。结论 蚌埠地区的乙型肝炎患者基因型以C型为主,B基因型次之,基因型与乙肝患者HBeAg阳性率、年龄等临床指标有一定的相关性。     

湖北省地区乙型肝炎病毒基因分型及其临床意义

目的探讨湖北省地区乙型肝炎病毒(HBV)基因型分布及其临床意义。方法随机选取HBVDNA阳性病例276例,其中慢性无症状携带者(其中30例经病理学证实)78例,慢性乙型肝炎110例,重症肝炎32例,肝硬化30例,肝细胞癌26例,取其外周血,采用乙型肝炎病毒核酸扩增荧光定量检测试剂盒检测HBVDNA载量,采用微板核酸杂交-ELISA法测定HBV基因型。结果湖北省地区基因型主要以C型和B型为主,基因型C型的HBVDNA载量均值水平为1.2×106copies/m l,显著高于其它类型,男女性别在HBV基因型构成比中无差异。混合型及基因型C与较严重的肝脏疾病有关,更易发生重症肝炎、肝硬化及肝癌。结论湖北省地区基因型主要以C型和B型为主,基因型C的HBVDNA载量均值水平显著高于其它类型,混合型及基因型C与较严重的肝脏疾病有关,湖北省地区存在基因型共同感染问题。     

呼和浩特市乙型肝炎病毒基因型分布及其临床特征

目的调查呼和浩特市乙型肝炎病毒(HBV)基因型分布情况及其临床特征。方法应用荧光定量PCR法对呼市128例慢性HBV感染者血清进行研究并分析不同基因型之间性别、年龄、临床诊断、HBV DNA水平、HBeAg状态、ALT及TBil水平的差异。结果呼市流行的HBV基因型主要为C型和非B非C型,其中C型占68.75%,非B非C型占26.56%,B型占4.69%。各基因型感染者年龄、性别、临床诊断、HBV DNA水平差异无统计学意义,HBeAg免疫状态及ALT、TBil水平差异有统计学意义。结论呼市流行的HBV基因型以C型为主,其次是非B非C型,少量为B型。     

肝癌患者血清中乙型肝炎病毒基因分型及临床意义

目的:探讨血清中乙型肝炎病毒(HBV)基因型及HBV DNA水平与肝细胞癌的关系。方法:应用巢式聚合酶链反应扩增乙型肝炎病毒与基因,用末端标记方法对PCR产物标记并直接测序,测序结果和GenBank中登录的标准基因型序列相比较,应用荧光定量PCR法检测HBV DNA水平。对61例肝癌、65例慢性乙型肝炎、10例乙型肝炎病毒携带者进行了检测。结果:136例中B基因型59例(43.4%)、C基因型77例(56.6%),随着病情加重,C基因型比例逐渐增高;不同基因型HBV感染的肝癌患者间HBV DNA水平差异有显著意义,P<0.05;在慢性乙型肝炎患者中,HBV DNA水平差异无显著性意义。结论:本地区乙型肝炎病毒以B、C基因型为主,乙型肝炎病毒C基因型及高水平的HBV DNA感染与肝癌的发生相关。     

乙型肝炎病毒基因型流行病学和临床意义

据世界卫生组织(WHO)估计,全世界约有20亿人感染过乙型肝炎病毒(HBV),其中的3亿5千万人成为病毒携带者.依据郭霍氏原则(Koch's postulate),微生物本身是造成疾病的一个决定因素;从遗传学而言,基因本身又决定了微生物的众多生物行为.     

乙型肝炎病毒基因分型与临床意义

乙型肝炎病毒（HBV）现已分为A～H8个基因型。应用HBV基因分型，研究其不同的临床特点，对HBV的防治、预后评估有一定的实用意义。我们对170例乙型肝炎（乙肝）患者进行HBV基因检测，分析其临床的相关性。     

秦皇岛市乙型肝炎病毒基因分型及分析

乙型肝炎病毒（HBV）从血清型到基因型的研究近年来取得很大的进展，不同地区基因型的分布及与临床及肝损害的关系不一致。我们对69例慢性乙型肝炎患者进行基因分型，其中部分病例进行肝穿活检。现报道如下。     

回族乙型肝炎病毒基因型分布及其临床意义

乙型肝炎病毒（HBV）分为8种不同的基因型。其型别与患者的病情严重程度及预后等有密切关系，其分布也存在一定的区域及民族差异。为进一步明确回族人群HBV基因型分布情况及其临床意义，进行了此项研究。     

乙型肝炎病毒基因型的研究现状及临床意义

乙型肝炎在全球广泛分布，慢性乙型肝炎是导致肝硬化、肝功能衰竭和原发性肝癌的主要危险因素，我国是HBV感染的高流行区。随着近几年分子生物学的发展，对HBV基因型的研究更加深入，基因型与临床的关系日趋明确，不同的基因型对乙型肝炎的发病机制、血清学诊断和抗病毒治疗对策有一定的影响。     

乙型肝炎病毒基因分型的标签序列研究

目的 快速、准确地鉴定HBV基因型,寻找基因型间特异性的序列标签.方法 通过生物信息学手段,比对分析美国国立生物技术信息中心数据库中930条各型HBV基因组序列的大S区域,选择型间具有高度特异性的序列片段作为分型标签序列,以此设计引物,通过PCR实验对重庆地区的80例临床HBV阳性血清样本进行分型观察. 结果 HBV基因组大S区的149～169 nt和461-483 nt两段区域含有多个保守的型间差异位点;在8种基因型标签对构成的28种型组合中,27种组合的碱基总差异数均≥6;PCR实验和电泳结果证实,来自该两区段的序列标签引物能够获得特异性的产物,有效鉴定出HBV基因型.结论 HBV基因大S区的149～169 nt和461～483 nt两段序列可作为HBV基因型分型标签序列,用作引物探针设计或测序标签.     

新疆维吾尔族慢性乙型肝炎患者HBV基因型分布及其特点

目的探讨新疆维吾尔族慢性乙型肝炎患者HBV基因型分布及其特点。方法采用型特异性引物巢式PCR法对127例维吾尔族慢性乙型肝炎患者进行基因分型,并测序验证。结果基因D型占39．4%（50/127）,基因B型占22．0%（28/127）,基因C型占16．5%（21/127）,基因BD混合型占9．4%（12/127）,基因CD混合型占8．7%（11/127）,基因BCD混合型占3．9%（5/127）; HBeAg阳性与HBeAg阴性的维吾尔族慢性乙型肝炎患者基因型分布,差异无统计学意义（x^2= 6．033,P＞0．05）;不同年龄维吾尔族慢性乙型肝炎患者HBV基因型分布差异无统计学意义（x^2= 3．137,P＞0．05）;不同性别维吾尔族慢性乙型肝炎患者HBV基因型分布差异亦无统计学意义（x^2= 8．058,P＞0．05）。结论新疆维吾尔族慢性乙型肝炎患者HBV基因型以D型占优势,其次可见B、C型及BD、CD、BCD混合型。同一疾病谱的慢性HBV感染者基因型分布可能与宿主HBeAg状态、年龄、性别无明显关系。     

乙型肝炎病毒三种基因分型方法的比较分析

随着乙型肝炎病毒（HBV）全基因序列测定病毒株的增多，近年人们开始根据生物系统发生学中的基因分类法建立HBV的基因分型技术和标准。对HBV进行基因分型有利于对HBV感染的流行病学、病因学和临床诊断与治疗进行更加深入细致的研究，现有研究表明不同亚型HBV感染的临床病程和对治疗的反应都存在一定的差异。我们分别应用自行建立的实时荧光聚合酶链反应（PCR）法、HBVS基因测序法和聚合酶链反应-限制性片段长度多态性（PCR—RFLP）法对75例慢性乙型肝炎患者的血清基因型进行检测，并对检测结果进行分析比较。     

Long-term clinical impact of occult hepatitis B virus infection in chronic hepatitis B patients

BACKGROUND/AIMS: Long-term clinical outcomes of occult hepatitis B virus (HBV) infection were studied. METHODS: Fifteen chronic hepatitis B patients were monitored for a median of 4.4 years (range 0.9-15.3) after hepatitis B surface antigen (HBsAg) seroclearance. Serum HBV DNA was measured by real-time detection polymerase chain reaction. Thirteen patients underwent liver biopsies at the end of follow-up and liver histology was evaluated by Ishak score. Liver HBV DNA was also measured for 12 patients. RESULTS: At the end of follow-up, HBV viremia was absent in 13 (87%) patients, and antibody titers to hepatitis B core antigen showed an inverse correlation with time from HBsAg seroclearance (r=-0.554; P=0.0040). However, all patients retained liver HBV DNA and tested positive for the covalently closed circular HBV DNA replicative intermediate. The hepatic HBV DNA loads had no relation to liver histology. Paired biopsies from 11 patients disclosed that each necroinflammatory score significantly improved after HBsAg seroclearance. Amelioration of liver fibrosis was also evident in eight (73%) patients (P=0.0391 by signed rank test). CONCLUSIONS: A long-standing but strongly suppressed HBV infection may confer histological amelioration after HBsAg seroclearance.     

Replication of clinical hepatitis B virus isolate and its application for selecting antiviral agents for chronic hepatitis B patients

AIM： To establish a cell model harboring replicative clinical hepatitis B virus （HBV） isolates and evaluate its application in individualized selection of anti-HBV agents for chronic hepatitis B （CHB） patients.
METHODS： The full-length HBV genomic DNA from 8 CHB patients was amplified by polymerase chain reaction （PCR）. All the patients were treated with lamivudine for at least seven months and finally became resistant to lamivudine. The amplified HBV DNA fragments were inserted into pHY106 vectors by Sap Ⅰ digestion. The recombinant plasmids containing 1.1 copies of HBV genome were transiently transfected into Huh7 cell line, and the levels of HBsAg, HBeAg and intercellular HBV replicative intermediates were determined by ELISA and Southern blot analysis, respectively, with or without lamivudine and adefovir treatment. The antiviral treatment with adefovir was administered to the patients and analyzed in parallel.
RESULTS： A total of 25 independent HBV isolates were obtained from the sera of 8 patients, each patient had at least two isolates. One isolate from each individual was selected and subcloned into pHY106 vector, including 5 isolates with YVDD mutation and 3 isolates with YIDD mutation. All recombinant plasmids harboring HBV isolates were transfected into Huh7 cells. The results indicated that HBV genome carried in HBV replicons of clinical HBV isolates could effectively replicate and express in Huh7 cells. Adefovir, but not lamivudine, inhibited HBV replication both in vitro and in vivo, and in vitro inhibition was dose-dependent.
CONCLUSION： The novel method described herein enables individualized selection of anti-HBV agents in clinic and is useful in future studies of antiviral therapy for CHB.     

Preliminary report of hepatitis B virus genotype prevalence in Iran

AIM: To determine the prevalence of hepatitis B virus (HBV) genotypes in Iranian hepatitis B surface antigen (HBsAg) carriers, chronic hepatitis B and cirrhotic patients. METHODS: A total of 109 HBsAg-positive patients were included in this study. HBV genotypes were determined by using INNO-LiPA methodology which is based on the reverse hybridization principle. RESULTS: The distribution of patients with different stages of liver disease was as follows: 95 (86.4%) chronic hepatitis, 11 (10%) liver cirrhosis, and 3 (2.7%) inactive carrier. Of the chronic hepatitis and liver cirrhosis patients, 26.4% were HBeAg-positive while 70% were HBeAg-negative. Genotype D was the only detected type found in all patients. CONCLUSION: Classifying HBV into genotypes has to be cost-effective and clinically relevant. Our study indicates that HBV genotype D prevails in the Mediterranean area, Near and Middle East, and South Asia. Continued efforts for understanding HBV genotype through international co-operation will reveal further virological differences of the genotypes and their clinical relevance.     

乙型肝炎病毒基因型(A-F)多引物PCR分型法的初步建立

目的：建立一种简便易行的乙型肝炎病毒基因分型方法（只需PCR）。方法:对GenBank中查获的114例HBV全序列进行比较分析，找出每种基因型相对于其他5种基因型的独特序列，并根据这些独特序列设计出6对分别针对A－F基因型的特异引物。用这6对引物分别对标本进行PCR，根据阳性结果判断出标本的基因型。进一步简化该方法，用多引物对PCR法（PCR with several primer sets in a single tube,Multiplex PCR)将B，C，D3种基因型的特异引物混合进行PCR，根据扩增片断的大小判断基因型。用此方法对已鉴定的B，C，D型标本进行比较和验证。结果：单引物对PCR与多引物对PCR的分型结果一致，且与以前用PCR－RFLP法的分型结果一致。结论：用多引物对PCR分型法准确易行，灵敏性高，便于推广应用。     

新疆地区汉族人群乙型肝炎病毒基因型的分布及其与临床类型的关系

本研究旨在通过型特异性引物巢式聚合酶链反应（PCR）的方法检测新疆地区汉族人群乙型肝炎病毒（HBV）患者HBV基因型并探讨与临床类型的关系，现将结果报道如下。     

Relationship between hepatitis B virus DNA levels and liver histology in patients with chronic hepatitis B

AIM： To study the relationship between hepatitis B virus （HBV） DNA levels and liver histology in patients with chronic hepatitis B （CHB） and to determine the prevalence and characteristics of hepatitis B e antigen （HBeAg） negative patients.
METHODS： A total of 213 patients with CHB were studied, and serum HBV DNA levels were measured by the COBAS Amplicor HBV Monitor test. All patients were divided into two groups according to the HBeAg status.The correlation between serum HBV DNA levels and liver damage （liver histology and biochemistry） was explored.
RESULTS： Of the 213 patients with serum HBV DNA levels higher than 10^5 copies/mL, 178 （83.6%） were HBeAg positive, 35 （16.4%） were HBeAg negative. The serum HBV DNA levels were not correlated to the age,history of CHB, histological grade and stage of liver disease in either HBeAg negative or HBeAg positive patients. There was no correlation between serum levels of HBV DNA and alanine aminotransferanse （ALT）,aspartate aminotrans-ferase （AST） in HBeAg positive patients. In HBeAg negative patients, there was no correlation between serum levels of HBV DNA and AST,while serum DNA levels correlated with ALT （r = 0.351, P = 0.042）. The grade （G） of liver disease correlated with ALT and AST （P 〈 0.05, r = 0.205, 0.327 respectively）in HBeAg positive patients. In HBeAg negative patients,correlations were shown between ALT, AST and the G （P 〈 0.01, and r = 0.862, 0.802 respectively）. HBeAg negative patients were older （35 ± 9 years vs 30 ±9 years, P 〈 0.05 ） and had a longer history of HBV infection （8 ± 4 years vs 6 ± 4 years, P 〈 0.05） and a lower HBV DNA level than HBeAg positive patients （8.4± 1.7 Log HBV DNA vs 9.8 ± 1.3 Log HBV DNA, P 〈0.001）. There were no significant differences in sex ratio,ALT and AST levels and liver histology between the two groups.
CONCLUSION： Serum HBV DNA level is not correlated to histological grade or stage of liver disease in CHB patients with HBV DNA mor