DNA, Circular ; isolation & purification ; DNA, Viral ; isolation & purification ; Hepatitis B ; diagnosis ; Hepatitis B virus ; genetics ; Humans

<b>OBJECTIVEb>To explore the variations of gene C in hepatitis B viruses between hepatitis B patients and healthy carriers, and provide experimental evidences for analysis of virus gene mutations acting on the virus material science and response of the body to the virus.

<b>METHODSb>The virus DNA load in hepatitis B patients and healthy blood donors was investigated by real-time polymerase chain reaction (PCR). Gene sequence analysis was taken to detect gene polymorphism, and all the success samples were compaired with standard strain by DNAstar.

<b>RESULTSb>(1)G Compared with standard strain, C region in all samples had mutations, there were 31 mutations in at least 2 samples (3 mutations in gene PreC and 28 mutations in gene C), including 9 missense mutations, 1 chain termination mutation and 21 synonymous mutation. Mutations nt 1827 c-->a and nt 2221 c-->t existed in all the samples, and most samples had 6 synonymous mutations. Four hepatitis B patients had mutation nt1896 g-->a, and another 4 patients had 2 mutations, namely, S87G and I97F (or 197L) in HBcAg CTL recognition episome. (2) The success ratio of amplification and sequencing of HBV DNA was closely associated with its copy numbers. In the present study, copy numbers of HBV DNA which were successfully amplified and sequenced were almost more than 40 193/ml.

<b>CONCLUSIONSb>HBV genome were easily affected by nucleotide mutations, 2 residues had mutations in gene of C region, which is firstly reported, suggesting these mutations may be geographical restricted. Mutations in gene of C region may either change the structure and function of HBeAg and HBcAg, which may further induce the escape of immune clearance for HBV or influence the detection of HBsAg or HBeAg, which may creat new problems for the prevention, diagnosis and treatment of hepatitis B.

Female ; Hepatitis B ; virology ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Mutation ; Polymorphism, Genetic

China ; epidemiology ; Hepatitis B ; classification ; epidemiology ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Mutation

Genes, Viral ; Genotype ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis B, Chronic ; virology ; Humans ; Polymerase Chain Reaction ; methods

Adult ; China ; epidemiology ; Genes, Viral ; Genotype ; Hepatitis B ; epidemiology ; virology ; Hepatitis B virus ; classification ; genetics ; isolation & purification ; Humans ; Male

<b>OBJECTIVEb>To identify the intergenotype recombination of hepatitis B virus (HBV) in Chinese patients and explore new recombination types of HBV.

<b>METHODSb>Complete genome sequences of HBV from Chinese patients were downloaded from GenBank and analyzed by Fragment-typing method to screen the potential recombinants, and the breakpoints of recombination were identified by using Simplot program.

<b>RESULTSb>Thirty-one recombination types including 755 recombinants were identified from 1642 complete HBV genome sequences, including 22 B/C recombination types with 676 recombinants, 5 C/D recombination types with 75 recombinants, 3 A/C recombination types with 3 recombinants, and 1 C/I recombination type with 1 recombinant.

<b>CONCLUSIONb>Of the 31 recombination types comprising 755 recombinants, 4 are novel recombination types found for the first time. All these recombination types identified in this study involve genotype C.

Genome, Viral ; Genotype ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Phylogeny ; Recombination, Genetic

<b>OBJECTIVEb>To investigate whether the hepatitis B virus (HBV) has quasispecies character by studying nucleotide sequence polymorphism and mutation features of HBV PreC/C gene region, and preliminaryly explore the heterogeneity of HBV quasispecies.

<b>METHODSb>The serum sample was obtained from a patient with chronic hepatitis B, and the whole HBV PreC/C gene region was amplified by PCR and cloned. Thirty-four clones that contained HBV PreC/C gene fragments were sequenced.

<b>RESULTSb>There were 28 kinds of different nucleotide sequences in 34 clones, and the nucleotide sequences diversity ranged from 0.2% to 2.1%. The mutation points were almost distributed in the whole region, but there wasn't mutation at PreC region nt.1 896 point in all sequences.

<b>CONCLUSIONb>Hepatitis B virus has complex quasispecies character in the patients with chronic hepatitis B.

Adult ; Hepatitis B Core Antigens ; genetics ; immunology ; Hepatitis B virus ; classification ; genetics ; isolation & purification ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Mutation ; Promoter Regions, Genetic ; genetics ; Sequence Analysis, DNA

<b>OBJECTIVESb>To study the phenomena of hepatitis B virus (HBV) integration into the tissues of hilar cholangiocarcinoma (HCCA) and to identify the integration sites in the host genome.

<b>METHODSb>Ten fresh HCCA samples were collected from the tissues by surgical ablation, 1 normal hilar bile duct sample selected as control. Cellular DNA were extracted by Wizard SV Genomic DNA Purification System. PCR-derived assay (HBV-Alu-PCR) was employed to amplify the viral-host junctions which contain the HBV sequence and the adjacent cellular flanking sequences. The PCR products were purified and subjected to sequencing by ABI-3730XL Auto DNA Analyzer. The sequence analysis of viral-host junctions was performed by DNASIS MAX 3.0 bioinformatics software. The insertion sites between viral and cellular sequences were identified through homology comparison using NCBI BLAST and MapViewer search.

<b>RESULTSb>In 10 HCCA samples, 5 were demonstrated to have HBV integration fragments with total 6 inserted sites identified. Sequence analysis from viral-host junction showed that HBV X gene inserted into host genome at random distribution with truncated fragments. HBV integration recurrently targeted the unknown region in upstream of CXXC finger protein-1 (CpG-binding protein) gene (4 cases). p53 tumor suppressor gene was also found at the integration site.

<b>CONCLUSIONSb>There is high integration rate of HBV DNA into cellular genome of HCCA. HBV integration is found frequently into or close to cancer-related genes. The findings demonstrate that HBV infection might have association with the pathogenesis of HCCA.

Aged ; Base Sequence ; Bile Duct Neoplasms ; genetics ; virology ; Cholangiocarcinoma ; genetics ; virology ; DNA, Viral ; genetics ; Female ; Hepatitis B ; virology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Virus Integration

<b>OBJECTIVEb>To know genotypes and serotypes of hepatitis B virus (HBV) detected from hepatitis B infected people in Yunnan Province.

<b>METHODSb>Serum samples were collected from HBsAg carriers detected from people who had a physical examination at Yunnan Provincial Center for Disease Control and Prevention. The S genes of HBV were amplified by nested PCR and the PCR products were sequenced. The viral genotype was identified by phylogenetic analysis. 27 reference sequences corresponding to HBV genotype A to I were obtained from GenBank. According to the amino acid sequences deduced from the nucleotide sequences of S gene, the dominant serotype of HBV detected from these people were confirmed.

<b>RESULTSb>39 HBsAg positive serum samples were detected from 2216 people who had a physical examination. The results shows that 76.9% were C genotype; 15.4% were B genotype; 5.1% were D genotype; 2.5% were I genotype. Three serotypes were found. The rates of adw2, adrq+ and ayr serotypes are 71.8%, 17.9% and 10.3% respectively. All of adw2 subtype specimens are C genotype. Among the serum specimens in which both HBsAg and HBeAg are positive, 75% were C genotype and adw2 subtype.

<b>CONCLUSIONb>It is determined that the main genotype and subtype of HBV prevailed in Yunnan province is C genotype and adw2 subtype.

Antibodies, Viral ; immunology ; China ; Female ; Hepatitis B ; virology ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B virus ; classification ; genetics ; immunology ; isolation & purification ; Humans ; Male ; Physical Examination ; Population Surveillance

<b>OBJECTIVEb>In order to investigate the characterization of mutation and genotype distributing in the younger group which was under the universal vaccination. The sequence of HBV was analyzed to offer the information to control and prevention in the area.

<b>METHODSb>Young person's sera with positive HBsAg are collected, and the Large S sequence of HBV including preS and S gene are amplified and sequenced. The genotype and serotype were determined by clastwal with the standard genotype sequence. And one virus complete genome is amplified.

<b>RESULTSb>The virus gene are successful amplified from the 33 sera. The sequence result indicate the 30 of 33 (90.9%) HBV genotype is B and 3 of 33 (9.0%) is C. The HBV serotype including ayw (1), adr (3), adw (29), 5 of 33 mutated in the "a" dominant of HBV, and the percentage is 15.2% . The HBV full length gene of serum number of 5856 is amplified and sequenced. Its genotype is B, serotype is adw and length is 3215 base.

<b>CONCLUSIONSb>The dominant genotype of HuNan is B, and the dominant serotype is adw.

Adolescent ; Child ; Child, Preschool ; China ; Female ; Genotype ; Hepatitis B ; virology ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; classification ; genetics ; isolation & purification ; Humans ; Male ; Mutation ; Phylogeny