Study on hepatitis C virus genotypes in Yantai district, Shandong province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To understand the hepatitis C virus (HCV) genotype distribution in Yantai district of Shandong province, and to explore whether the HCV genotypes was relevant to the injure of liver through the index of liver function.</p><p><b>METHODS</b>Using specific PCR primers to amplify the HCV RNA 5' UTR/NS5B,then PCR products were sequenced by genetic analyzer. The genotypes were identified by alignment to the GenBank reference sequences and construction the phylogenetic tree of 5' UTR.</p><p><b>RESULTS</b>Among 9 unpaid blood donors we detected two kinds of genotypes of 1b and 3a, respectively, 8 cases (88.9%) and 1 case (11.1%). Among 33 cases of hepatitis C patients we detected the 1b, 2a and 6a the three kinds of genotypes, respectively, 22 (66.7%), 10 (30.3%) and 1 (3.03%) cases. Subtype 1b is the advantage of popular genotype in HCV carriers from Yantai district, and the distribution of 1b was no significant difference in the different population (chi2 = 0.796, P = 0.373); The difference of indicator of liver damage in the different genotypes of subjects were significant (P < 0.05), the mean of ALT, AST of 2a-subtype carriers was significantly higher than the 1b-subtype population.</p><p><b>CONCLUSIONS</b>The genetic diversity of HCV in Shandong Yantai district was presented. The main genotypes were 1b-subtype, and 3a and 6a-subtype was detected firstly. The genotype of HCV were relevant to the liver damage indicators, 2a-subtype hepatitis C virus infection in the liver cells may play an important role in the disease process.</p>

Detection and analysis of gene polymorphism in hepatitis B virus C region / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>(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.</p><p><b>CONCLUSIONS</b>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.</p>

Polymorphisms of Epstein-Barr virus BHRF1 gene, a homologue of bcl-2 / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>EBV BamHI fragment H rightward open reading frame 1 (BHRF1), the Epstein-Barr virus (EBV) early gene, is structurally and functionally homologous to the oncogene bcl-2 and may play an important role in the development of EBV-associated tumors. To characterize the polymorphisms of BHRF1 in EBV-associated tumors, we analyzed the sequences of BHRF1 in isolates from nasopharyngeal carcinoma (NPC) and EBV-associated gastric carcinoma (EBVaGC) biopsies as well as throat washing (TW) samples from healthy donors.</p><p><b>METHODS</b>BHRF1 DNA sequences were analyzed by polymerase chain reaction (PCR) and sequencing for 39 NPC samples, 40 EBVaGC samples, and 53 EBV-positive TW samples from healthy donors. The variants of BHRF1 gene were classified according to the signature changes. The EBV types 1 and 2 at nuclear antigen (EBNA) 3C locus were determined by PCR.</p><p><b>RESULTS</b>Compared with EBV standard cell line B95-8, all isolates carried a silent mutation at amino acid (AA) 80 (nucleotide 54616 T→C), the AA88 L→V mutation was found in most isolates, and the AA79 V→L mutation in a few isolates. Other mutations were sporadically distributed. Based on the mutations at AA88 and AA79, 3 distinct variants of BHRF1 genes, designated as 79V88V, 79L88L, and 79V88L, were identified. The 79V88V was the most common variant. The distribution of the BHRF1 variants among the NPC, EBVaGC, and TW samples was not significant. The corresponding regions of bcl-2 homologues were conserved in all isolates except for 3 samples. The distribution of BHRF1 variants in type 1 and type 2 strains was significant different (P < 0.001, contingency coefficient was 0.554).</p><p><b>CONCLUSIONS</b>The 79V88V is the dominant variant in NPC, EBVaGC, and TW samples from healthy donors and preferential linkages between BHRF1 and EBNA3C variants exist. Conserved BHRF1 in Bcl-2 homologous domains is helpful to remain the important role of BHRF1.</p>