青海同德藏族人群感染HBV基因型、血清型调查

目的 了解青海同德藏族人群乙型肝炎病毒(HBV)基因型、血清型的分布状况.方法 用巢式PCR扩增青海同德藏族人群表面抗原(HBsAg)阳性者的HBV S、C基因,测定其序列,判定其基因型和血清型.结果 源于自然人群的311份血清标本中,同时获得S和C基因序列者271份,其中C型基因10例,占3.7%,C/D重组型261例,占96.3%;血清ayw2亚型259例,占95.6%,adr亚型10例,占3.7%,adw2亚型2例,占0.7%.结论 青海同德藏族人群感染HBV以C/D重组型为主,血清型以ayw2为主.     

云南省乙型肝炎病毒基因型和血清型的流行状况分析

目的 了解云南省人群中流行的乙型肝炎病毒(Hepatitis B virus,HBV)基因型和血清型分布情况.方法 从参加健康体检的人群中筛查HBsAg阳性的血清标本,利用巢式PCR扩增HBV S基因片段,并对扩增产物进行序列测定.从GenBank中查获A～I基因型共27株HBV参考序列,构建HBV S基因系统发生树,以此确定其样本的基因型及血清型,并对结果进行统计分析.结果 从2216名体检人员血清标本中筛查出 39份HBsAg阳性的样本,HBV的S基因序列分型结果表明有4种基因型:C型占76.9%,B型占15.4%;D型占5.1%;I型占2.5%.血清分型结果为:adw2型占71.8%;adrq+型占17.9%;ayt型占10.3%.所有adw2血清型标本均为C基因型.HBsAg、HBeAg双阳性标本中75%为C基因型/adw2血清亚型.结论 云南省HBV感染人群中HBV基因型的分布以C型为主,血清型以adw2型为主.     

云南省乙型肝炎病毒基因型和血清型的流行状况分析

目的 了解云南省人群中流行的乙型肝炎病毒(Hepatitis B virus,HBV)基因型和血清型分布情况.方法 从参加健康体检的人群中筛查HBsAg阳性的血清标本,利用巢式PCR扩增HBV S基因片段,并对扩增产物进行序列测定.从GenBank中查获A～I基因型共27株HBV参考序列,构建HBV S基因系统发生树,以此确定其样本的基因型及血清型,并对结果进行统计分析.结果 从2216名体检人员血清标本中筛查出 39份HBsAg阳性的样本,HBV的S基因序列分型结果表明有4种基因型:C型占76.9%,B型占15.4%;D型占5.1%;I型占2.5%.血清分型结果为:adw2型占71.8%;adrq+型占17.9%;ayt型占10.3%.所有adw2血清型标本均为C基因型.HBsAg、HBeAg双阳性标本中75%为C基因型/adw2血清亚型.结论 云南省HBV感染人群中HBV基因型的分布以C型为主,血清型以adw2型为主.     

乙肝疫苗长期免疫地区低龄人群乙型肝炎病毒基因序列与突变现状

目的 通过乙型肝炎流行病学调查和病毒基因检测,了解乙肝疫苗长期免疫地区低年龄人群的基因序列与突变特征.方法 从乙肝监测点收集乙型肝炎病毒表面抗原阳性血清,取其中年龄小于16岁者,扩增包括preS和S基因在内的基因序列片段,共1100碱基,序列测定后与标准基因型别比较,确定病毒基因及血清型别,确定a抗原决定簇的氨基酸替代突变发生率,选取一株病毒进行全基因扩增、克隆和序列测定.结果 共检测样本35例,33例血清扩增出乙肝病毒基因序列,其中30例乙型肝炎病毒基因型为B型,占90.9%,3例乙型肝炎病毒基因型为C型,占9.0%,1例血清型为ayw,3例为adr,其余29例为adw,共有5例在a抗原决定簇发生氨基酸替代突变,发生率约为15.2%.其中5856号血清扩增乙型肝炎病毒全基因为B型,血清型为adw,共3215个碱基.结论 该地区乙型肝炎基因型主要为B型,血清型为adw,人群中a抗原决定簇某些位点已发生与疫苗免疫逃逸相关的突变.     

云南省乙型肝炎病毒基因型和血清型的流行状况分析

目的了解云南省人群中流行的乙型肝炎病毒（Hepatitis B virus,HBV）基因型和血清型分布情况。方法从参加健康体检的人群中筛查HBsAg阳性的血清标本,利用巢式PCR扩增HBVS基因片段,并对扩增产物进行序列测定,从GenBank中查获A～I基因型共27株HBV参考序列,构建HBVS基因系统发生树,以此确定其样本的基因型及血清型,并对结果进行统计分析。结果从2216名体检人员血清标本中筛查出39份HBsAg阳性的样本,HBV的s基因序列分型结果表明有4种基因型：c型占76．9％,B型占15．4％;D型占5．1％;I型占2．5％。血清分型结果为：adw2型占71．8％;adrq‘型占17．9％;ayr型占10．3％。所有adw2血清型标本均为c基因型。HBsAg、HBeAg双阳性标本中75％为c基因型／adw2血清亚型。结论云南省HBV感染人群中HBV基因型的分布以C型为主,血清型以adw2型为主。     

肝癌高发区广西隆安县乙型肝炎病毒株全基因序列分析

目的 探讨肝癌高发区广西隆安县乙型肝炎病毒(HBV)株的全基因序列是否存在地域特殊性。方法 用套式PCR(nPCR)扩增该县HBV无症状携带者血清HBV全基因，用克隆测序法测序并做同源性对比。结果 本病毒株共3215个碱基，基因型为C，血清型为adw。其P基因区有40处发生碱基点突变，导致11个氨基酸改变；S基因区的前S1、前S2和S基因分别有11、2和3个碱基点突变，分别导致3、1、1氨基酸改变，未发现“a”区突变；X基因区共有6个碱基点突变，导致4个氨基酸改变，其中出现能影响e抗原表达的双突变(nt1762A→T、1764G→A)；未发现前C区基因突变，C基因有13个碱基发生突变，导致2个氨基酸改变。本病毒株与越南北部病毒株高于同为C基因型的上海株、北京株、西藏株。结论 未发现肝癌高发区的本例HBV基因C型的隆安株，基因虽有肝癌高发性，但无地域特殊性。     

乙型肝炎病毒B/C基因型重组新方式的探讨

目的 研究HBV B/C基因型重组方式.方法 采用聚合酶链反应扩增4例景洪僾伲族慢性乙型肝炎患者血清中的HBV全基因并与pMD18-T载体相连,转化入大肠杆菌E.coli JM109中,经限制性内切酶酶切鉴定,阳性克隆DNA序列测定后进行HBV基因分型及重组部位的鉴定.结果 4名患者16条HBV全基因均为有不同程度的C基因型毒株重组的B基因型,重组方式有两种:群1与C基因型毒株重组部位只有1个,位于HBV前C/C区nt1825～nt2320,共496 bp;群2与C基因型毒株重组部位有2个,位于P基因区nt822～nt1020和前C/C基因区nt1825～nt2320,共695 bp.结论 这种B/C重组方式尚未见报道,Bj亚型与C基因型毒株除了在前C/C基因区重组外,同时还伴有少部分P基因区重组.     

乙型肝炎病毒基因组前-前-S和前-X区基因分布的研究

目的研究不同基因型乙型肝炎病毒(HBV)前-前-S和前-X区核苷酸和氨基酸序列分布。方法应用DNAstar软件中的MegAlign程序分析GenBank登记的35株和3株由本室测定的不同基因型HBV全序列中前-前-S和前-X区基因分布情况，并用Blast软件对GenBank中598株HBV全基因组序列进行前-前-S和前-X区的核苷酸和氨基酸相似性分析。结果除D基因型外，在各型HBV基因组中均存在前-前-S区核苷酸序列，但仅在C、F和H型HBV基因组中存在前-前-S区开放读码框架(ORF)。但前-X区核苷酸序列及其ORF仪见于C基因型HBV。在GenBank登记的598株不同基因型HBV中，与推导的前-前-S区相似氨基酸序列有36株，而与推导的前-X区相似的氨基酸序列有47株。结论前-前-S区序列存在于除D基因型以外的所有HBV基因型，而前-X区基因仅存在于C基因型HBV基因组中。     

检测HBV基因组nt585位突变的特异性聚合酶链反应

目的　为调查乙型肝炎病毒(HBV)nt585位A→C变异株在国内的流行情况提供方法学。方法　根据已知的HBV基因组序列并结合国内主要流行adr和adw亚型的特点,合成在nt585位分别为A和C的2套4对引物,建立了突变特异性聚合酶链反应(msPCR)方法。结果　利用msPCR对由25份乙型肝炎免疫失败儿童患者血清和32份成人乙型肝炎患者血清扩增的HBVS基因片段进行了初步检测。结果表明该法可特异性扩增nt585为A和C的HBVDNA。结论　该法是鉴定HBV基因组nt585位A→C变异株的特异而敏感的方法。     

HBV前C基因突变及分型与HBV母婴传播关系的研究

目的探讨HBV(乙型肝炎病毒)前C基因突变、血清型及基因型分型与HBV母婴传播的关系。方法100份HBsAg阳性孕产妇及其新生儿的外周血,发生母婴传播血清为病例组,其他为对照组。ELISA法检测乙肝两对半,根据S基因判断HBV血清型和基因型。对HBV前C基因进行扩增及序列分析。结果99份HBV血清型全为adw型;39例发生母婴传播HBV基因C型25例;B型13例;E型1例。60例未发生母婴传播,HBV基因B型50例;C型10例。两组HBV基因分型结果差异有统计学意义(P0.05)。39例发生母婴传播中27例发生了点突变,12例无任何突变,突变发生率为69%。结论HBV母亲C基因型可能更易发生母婴传播。HBV母婴传播中大多存在HBV前C基因变异并导致氨基酸取代,其中主要涉及丝氨酸、苏氨酸、酪氨酸等的磷酸化位点,这些重要位点氨基酸的取代可能在HBV母婴传播的发病中起作用。     

Genotypes and phylogenetic characterization of hepatitis B and delta viruses in Egypt

Hepatitis B virus (HBV) and hepatitis D virus (HDV) sequences among HBV carriers from Egypt have not been evaluated sufficiently. The genotypes of HBV isolated from 105 serum samples from Egyptian carriers were determined. Four complete genomes and 11 entire preS1/S2/S genes were sequenced and evaluated. All serum samples were classified into HBV genotype D using serologic and genetic methods. The length of four complete nucleotide sequences was 3,182 bp. In all 15 samples, the common 33 nucleotides (11 amino acids) deletions in the preS1 region specific for HBV genotype D were observed. In the phylogenetic analysis based on the complete nucleotide sequences, all samples were clustered with the HBV isolates reported from previously Western and Mediterranean countries with nucleotide homology ranging from 96.0-98.0%. Of 75 HBsAg positive samples, anti-HDV was found in 15 (20%), and HDV RNA was detected in 9 of 15 (60%). The proportion of the patients with liver disease was higher in HBV carriers of anti-HDV positive with HDV RNA than in HBV carriers of anti-HDV positive without HDV RNA (P < 0.05). In the phylogenetic analysis based on the sequences in nucleotide position 853-1267 of HDV, nine samples were classified into HDV genotype I with the nucleotide homology ranging from 88.3-92.1% (mean; 90.5%) and clustered with HDV strains reported previously from Ethiopia, Somalia, Egypt, and Lebanon. These results indicate that HBV genotype D and HDV genotype I are most prevalent in Egypt, and HDV co-infection in HBV carriers is related to severity of liver disease.     

河南省漯河地区乙型肝炎病毒基因型别的初步研究

目的 初步确定流行于河南漯河地区的乙肝病毒的基因型的基本情况.方法 采集河南漯河地区乙型肝炎表面抗原阳性的血清,利用PCR扩增得到HBV的S基因;用DANSTAR软件对其进行核苷酸序列分析,构建系统发生树,分析基因型.结果约75.7%样本的HBV S基因序列位于HBV系统发生树的基因型C,约20%的样本其S基因位于系统发生树的基因型B,近4.3%的样本其S基因位于系统发生树的基因型D.结论 流行于河南漯河地区的乙肝病毒多数为基因型C,少数为基因型B,极少数为基因型D.     

Genotype,serotype, and phylogenetic characterization of the complete genome sequence of hepatitis B virus isolates from Malawian chronic carriers of the virus

Hepatitis B virus (HBV) genotypes have distinct geographical distribution. HBV sequences among hepatitis B carriers in Malawi have not been evaluated thus far. HBsAg serotype and genotype of HBV was determined in 20 serum samples from Malawian chronic HBV carriers, and two complete genomes and 13 entire pre-S2/S genes were sequenced directly. Genotype A HBV isolates were found in all of the samples, and serotype with adw2 and ayw2 were detected in three and 17 samples, respectively. In phylogenetic analyses, two complete genomes were classified into a subgroup A' that was described previously in South African isolates of the virus, and were separated from HBV isolates in Western countries with nucleotide differences ranging from 4.1-6.2%. The separation of subgroup A' was also evident in the tree topology of the entire pre-S1/S2, X and precore/core region, but not evident in the small-S region. The nucleotide divergences in subgroup A' were higher than those among genotype A without subgroup A' in the complete genomes as well as each of four open reading frames. All of the 13 pre-S2/S sequences were classified into the subgroup A', and clustered with known HBV isolates with ayw2 in carriers from South Africa and Zimbabwe. Three amino acids in the pre-S2/S gene were characteristic of subgroup A' with ayw2. In conclusion, unique HBV isolates of subgroup A' with ayw2 are prevalent in Malawi, and subgroup A' with a relatively higher nucleotide diversity may be a HBV isolate characteristic of the indigenous population of some African countries.     

Phylogenetic analysis of hepatitis B virus genotype F complete genome sequences from Chilean patients with chronic infection

Molecular epidemiological data concerning the hepatitis B virus (HBV) in Chile are not known completely. Since the HBV genotype F is the most prevalent in the country, the goal of this study was to obtain full HBV genome sequences from patients infected chronically in order to determine their subgenotypes and the occurrence of resistance-associated mutations. Twenty-one serum samples from antiviral drug-naive patients with chronic hepatitis B were subjected to full-length PCR amplification, and both strands of the whole genomes were fully sequenced. Phylogenetic analyses were performed along with reference sequences available from GenBank (n = 290). The sequences were aligned using Clustal X and edited in the SE-AL software. Bayesian phylogenetic analyses were conducted by Markov Chain Monte Carlo simulations (MCMC) for 10 million generations in order to obtain the substitution tree using BEAST. The sequences were also analyzed for the presence of primary drug resistance mutations using CodonCode Aligner Software. The phylogenetic analyses indicated that all sequences were found to be the HBV subgenotype F1b, clustered into four different groups, suggesting that diverse lineages of this subgenotype may be circulating within this population of Chilean patients.     

山西省乙型肝炎病毒基因型别的初步研究

目的 初步确定流行于山西地区的乙肝病毒的基因型的基本情况。方法 采集山西地区乙型肝炎表面抗原阳性的血清，利用PCR扩增得到HBV的S基因和C基因；用MEGA3软件对其进行核苷酸序列分析，构建系统发生树，分析基因型。结果 约93％样本的HBVS基因和C基因序列均位于HBV系统发生树的基因型C，近7％的样本其S基因和C基因序列位于系统发生树的基因型B。结论 流行于山西地区的乙肝病毒多数为基因型C，少数为基因型B，未发现基因重组现象。     

Comparison of genotypes C and D of the hepatitis B virus in Japan: a clinical and molecular biological study

The genotype-related differences between genotype C and genotype D of the hepatitis B virus (HBV) remain unknown. The relationship was studied between the HBV genotypes and their clinical features, paying special attention to genotypes C and D. Serum samples from 413 HBV carriers were genotyped using an enzyme immunoassay (EIA) and by restriction fragment length polymorphism. The nucleotide sequences at the basic core promoter (BCP) and precore (PreC) regions were analysed by direct sequencing. The full genome sequences of three HBV genotype D cases were also examined. Almost all carriers with HBV genotype D were asymptomatic carriers (84.2%). Genotype D was not found in patients with liver cirrhosis and hepatocellular carcinoma. In contrast, carriers with genotype C had mainly chronic liver disease (63.2%; P<0.001). The ratio of hepatitis B e antigen (HBeAg)/anti-HBe was significantly higher in genotype C than in genotype D in the young age-matched group (P<0.01). The mutation at BCP (T1762, A1764) was significantly lower in genotype D than in genotype C among HBeAg-negative patients (P<0.05). The HBV full-genome sequences are very similar to certain HBV genotype D sequences from Europe. In conclusion, genotype C was associated with chronic liver disease, whereas genotype D was related to asymptomatic carriers with earlier HBeAg seroconversion. Thus, the outcome of chronic HBV infection may be different in persons infected with HBV genotypes C and D.     

Genotype characterization and phylogenetic analysis of hepatitis B virus isolates from Iranian patients

Hepatitis B virus (HBV) is one of the major causative agents of acute and chronic liver disease worldwide and is believed to be responsible for a million deaths annually. Eight genotypes of HBV, A to H, have been described on the basis of similarity of the complete genomes sequence. Although, it is reported that the predominant HBV genotype in the Mediterranean area and the middle east is genotype D, there are no reports on HBV genotypes prevalent in Iran. In this study, the C and S regions of HBV from 26 chronic hepatitis B Iranian patients were amplified and sequenced. Phylogenetic analysis revealed that all Iranian HBV isolates sequences were classified into genotype D with bootstrap values of 100%, 73%, and 100% (1,000 replicates each) for S, C, and preS2 regions, respectively. The mean percent intra-distance of S and C regions were 0.8% and 2.3%, respectively. The mean percent inter-distance of S and C regions between Iranians and genotype D isolates were 1.7% and 3.0%, respectively, and the range of mean percent nucleotide distance of S and C regions between Iranians and the other reference isolates were 7.9%-17.5% and 4.8%-14.7%, respectively. Thirteen out of 23 HBV C region sequences showed nucleotide "A" at position 1896 (precore mutant) in C region. Nucleotide 1858 showed presence of "T" in all isolates. No insertion or deletion was found in both regions. SimPlot and BootScanning analyses did not show any recombination between Iranian isolates and other genotypes in both regions.     

Novel hepatitis B virus subgenotype A6 in African-Belgian patients

BackgroundGenome diversity of hepatitis B virus (HBV) is prominent among DNA viruses; which, allowed the virus to be genetically classified into eight genotypes and several subgenotypes.ObjectiveTo introduce and to characterize a novel subgenotype HBV, classified as A6.Study designHBV full-length genomes were isolated and sequenced from three African-Belgian patients chronically infected with the virus. Using phylogenetic reconstruction and genetic distance calculation, the evolutionary relationships of the novel strains were investigated.ResultsPhylogenetic analysis based on complete genome sequences of genotype A strains revealed distinct clusters supported by high bootstrap values. The three African-Belgian strains clustered separately from the other known A subgenotypes (A1–A5) with maximal bootstrap support (100%). The mean inter-subgenotypic nucleotide divergence over the complete genome sequence between the novel A6 strains and A1–A5 was higher than 4%.ConclusionPhylogenetic analysis of the complete genome sequences yielded maximal bootstrap value support for nodes that establish the new lineage as a novel subgenotype. In addition, nucleotide divergence more than 4% based on full-length genome of the virus, clearly demonstrated that the three African-Belgian strains belonged to a novel subgenotype of HBV, which was assigned as “A6”. Noteworthy, the phylogeny of genotype A demonstrated that the A6 is a basal lineage that diverged earlier from the other African subgenotypes of genotype A.     

A new isolate of hepatitis B virus from the Philippines possibly representing a new subgenotype C6

Hepatitis B virus (HBV) genotypes and subgenotypes show distinct geographical prevalence. A genotyping analysis of 28 samples from asymptomatic HBV carriers from the Philippines gave a distribution of HBV genotypes as expected from a previous study: 54% B (15/28), C5 18% (5/28), 14% D (4/28), 7% A1 (2/28). In addition, 7% (2/28) of the samples showed a double infection with genotypes B and D. One of the isolates sequenced completely, ph105, did not group into one of the known subgenotypes after phylogenetic analysis. Ph105 formed a separate clade in genotype C. With a genome length of 3,215 nt. and a serological subtype adr, ph105 exhibited the main features of most genotype C strains. However, ph105 differed by 4.1–7.2% from HBV subgenotypes C1 to C5 when comparing the nucleotide sequence of whole genomes. With only 4.1% difference ph105 was most closely related to subgenotype C2. SimPlot analysis gave no indication for recombination with known HBV genotypes. Ph105 fulfils all criteria for a new subgenotype C6. J. Med. Virol. 81:983–987, 2009. © 2009 Wiley‐Liss, Inc.     

沈阳地区乙型肝炎病毒基因型分子流行病学研究

目的 研究沈阳地区乙型肝炎病毒基因型分布和特点。方法 应用半巢式聚合酶链反应（PCR）扩增乙型肝炎病毒P基因，将PCR产物应用ABI377自动测序仪直接做核苷酸序列分析，并用DNA STAR软件进行种系发生分析及基因型鉴定。结果 HBV DNA标准株P基因片段可进行基因分型。在沈阳地区慢性HBV感染者中可检出基因型B、C和D，检出率分别为22％、50％和28％，基因型C分布与基因型B、D的差异有统计学意义，在慢性乙型肝炎患者和慢性HBV携带者间各基因型间分布比较中差异无统计学意义。结论 通过测定HBV DNAP基因序列片段可进行HBV DNA基因分型。沈阳地区乙型肝炎病毒基因型有基因型B、C和D型，其中基因型C为优势基因型。