OBJECTIVETo study hepatitis B virus (HBV) genotype and subtype distribution and its clinical significance in HBV-infected patients.

METHODSWe used type/subtype-specific primers and PCR to detect HBV genotype and subtype of 445 HBV-infected patients from Beijing, Changchun, Hanchuan Shenzhen, Qingyuan and Nanjing, including 7 acute hepatitis (AH), 36 asymptomatic HBV carriers (ASC), 352 chronic hepatitis (CH), 28 liver cirrhosis (LC), and 22 hepatocellular carcinoma (HCC) cases. Genotyping results were confirmed by PCR product sequencing.

RESULTSAmong 445 HBV-infected patients, the proportions of genotype B, C, and B/C were 32.6% (145/445), 53.7% (239/445), and 13.7% (61/445), respectively. In genotype C, 13 (5.4%) were subtype C1, 135 (56.5%) were subtype C2, and the remaining 91 (38.1%) were neither C1 nor C2. In genotype B, 100 (69.0%) were subtype Ba, 25 (17.2%) subtype Bj, and the other 20 (13.8%) were neither Ba nor Bj. In genotype B/C, 15 (24.6%) were Ba/C2, 8 (13.1%) Bj/C2, 6 (9.8%) Ba/C1, 3 (4.9%) Bj/C1, 11 (18.0%) Ba/neither C1 nor C2, 7 (11.5%) Bj/neither C1 nor C2, and 6 (9.8%) neither Ba nor Bj/neither C1 nor C2, 2 (3.3%) neither Ba nor Bj/C1, 3 (4.9%) neither Ba nor Bj/C2. The HBV genotype and subtype distribution we found exhibited significant differences in the various clinical types of HBV infection tested, and showed that genotype C was predominant among patients with liver cirrhosis (78.6%) and hepatocellular carcinoma (86.4%) while genotype B was predominant in asymptomatic carriers (72.2%). In addition, genotype and subtype distribution showed no significant differences between male and female patients, but genotype and subtype distribution showed significant differences in patients positive or negative with HBeAg.

CONCLUSIONSubtypes Ba and C2 are predominant in patients with hepatitis B from these 6 cities, and genotype C may be associated with the development of liver cirrhosis and hepatocellular carcinoma.

Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Liver Cirrhosis ; virology ; Liver Neoplasms ; virology ; Polymerase Chain Reaction

BACKGROUND/AIMS: The genotype of hepatitis B virus (HBV) has been suggested to correlate with the clinical outcome of HBV infection. We analyzed the distribution of HBV genotypes according to the clinical outcomes of HBV infection in Jeju island. METHODS: A total of 145 HBsAg-positive samples were enrolled. To identify specific patterns of HBV genotypes, we performed restriction fragment length polymorphism (RFLP). In the case that typical restriction pattern of RFLP was not determined, phylogenetic analysis was performed. RESULTS: RFLP analysis was possible in 65 patients. Clinical diagnoses of the 65 patients with chronic liver disease were as follows: HBeAg-positive healthy carrier (HBeAg+, anti-HBe, HBV DNA+, transaminase normal; n=20); Inactive HBsAg carrier (n=12); chronic hepatitis B (n=14); liver cirrhosis (n=9); hepatocellular carcinoma (n=10). Sixty-two patients showed a typical restriction pattern by HinfI. However, 3 patients showed a unique restriction pattern by HinfI, which were not reported in the literature. When phylogenetic analysis was performed to classify the genotype of these 3 patients, they were also genotype C. However, all 65 patients showed typical restriction patterns by Tsp509I, which were reported in genotype C. CONCLUSIONS: All chronic HBV infections are genotype C in Jeju island regardless of clinical outcomes.
Carcinoma, Hepatocellular/virology ; Carrier State/virology ; Genotype ; Hepatitis B virus/*genetics ; Hepatitis B, Chronic/complications/*virology ; Humans ; Liver Cirrhosis/virology ; Liver Neoplasms/virology ; Polymorphism, Restriction Fragment Length

Carcinoma, Hepatocellular ; virology ; DNA, Viral ; metabolism ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Liver Cirrhosis ; virology ; Liver Neoplasms ; virology ; Virus Integration ; physiology

OBJECTIVETo investigate the distribution of hepatitis B virus genotype in Hubei province (China) and its clinical significance.

METHODSSerum samples from 190 HBV DNA positive patients with chronic HBV infection,including 52 asymptomatic HBV carriers (ASC), 56 chronic hepatitis (CH), 32 fulminant hepatic failure (FHF), 22 liver cirrhosis (LC), and 28 hepatocellular carcinoma (HCC) patients were collected and tested for HBV genotypes by type-specific primers.

RESULTSA simple and precise genotyping system based on PCR using type-specific primers was developed for the determination of genotypes of hepatitis B virus (HBV). Of the 190 patients, 140 (73.7%) were genotype B and 42 (22.1%) were genotype C. Genotype B was more prevalent in the FHF and HCC patients than in the ASC patients; the ALT value was significantly higher in genotype B than in genotype C patients. The rate of anti-HBe was significantly higher in genotype B than in genotype C except in the patients of the ASC group.

CONCLUSIONThe system we used seems to be a useful tool for the molecular diagnosis of HBV infection and for large-scale surveys. Genotype B, genotype C and BC combination exist in Hubei province, and genotype B is the major genotype in this area especially in FHF and HCC patients.

Adult ; Carcinoma, Hepatocellular ; virology ; Carrier State ; virology ; China ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; complications ; virology ; Humans ; Liver Cirrhosis ; virology ; Liver Failure, Acute ; virology ; Liver Neoplasms ; virology ; Male ; Middle Aged

Adolescent ; Adult ; Asian Continental Ancestry Group ; Carcinoma, Hepatocellular ; genetics ; virology ; Ethnic Groups ; Female ; Genotype ; Hepatitis B ; genetics ; virology ; Hepatitis B virus ; genetics ; Humans ; Liver Cirrhosis ; genetics ; virology ; Liver Neoplasms ; genetics ; virology ; Male ; Middle Aged ; Mutation ; Young Adult

The mortality due to chronic liver disease, including liver cirrhosis and hepatocellular carcinoma (HCC), ranks as one of the highest in Korea. The prevalence rates of hepatitis C virus (HCV) and hepatitis B virus (HBV) infections in the general Korean population are approximately 1 and 5%, respectively. Blood transfusion was the strongest risk factor for the transmission of HCV infection. Therefore, the evaluation of risk factors for HCV infection including blood transfusion, intravenous drug user, hemophilia, and hemodialysis, is important. The most prevalent HCV genotype is 1b followed by 2a. The annual incidence of HCC among HCV-related liver cirrhosis has been estimated at 5%, and approximately 12% of HCC is attributable to HCV and 68% to HBV in Korea. HCV infection is more closely associated with HCC in elderly patients than HBV-related HCC. Even though the prevalence of anti-HCV in Korea has been reduced and the risk of HCV transmission through blood transfusion has markedly decreased, public-health programs to prevent de novo infections should be developed. This review describes the HCV prevalence and risk factors among the general population, and the distribution of HCV genotypes as well as the clinical course of HCV in Korea.
Adult ; Carcinoma, Hepatocellular/*virology ; Genotype ; Hepacivirus/genetics/isolation & purification ; Hepatitis C, Chronic/*complications/*epidemiology ; Humans ; Korea ; Liver Cirrhosis/*virology ; Liver Neoplasms/*virology ; Middle Aged ; Prevalence ; Risk Factors

OBJECTIVETo investigate the association of genetic polymorphisms in glutathione S-transferases(GST) M1 with hepatitis beta-related hepatocellular carcinoma (HCC).

METHODSGenomic DNA was isolated from peripheral blood of HBsAg carriers, including 91 cases of HCC, 58 liver cirrhosis(LC), 63 chronic hepatitis B(CHB), and 134 normal controls. GSTM1 genotypes were detected by multiplex PCR.

RESULTSThe null genotype of GSTM1 was significantly frequent in patients with HCC compared with controls (P<0.05), but there were no significant differences in frequency of GSTM1 null genotype among patients with liver cirrhosis, chronic hepatitis B and normal controls. Subjects carrying null genotypes of GSTM1 had higher risk of developing HCC compared with those carrying positive genotype (OR=1.81.95% CI=1.05 approximately equals 3.12).

CONCLUSIONThe GSTM1-null genotype may be associated with an increased risk of HCC, but not of CHB and LC.

Adult ; Carcinoma, Hepatocellular ; etiology ; genetics ; virology ; Female ; Genotype ; Glutathione Transferase ; genetics ; Hepatitis B, Chronic ; complications ; genetics ; Humans ; Liver Cirrhosis ; etiology ; genetics ; virology ; Liver Neoplasms ; etiology ; genetics ; virology ; Male ; Middle Aged ; Polymorphism, Genetic

OBJECTIVETo construct a cDNA library from human liver tissue of cirrhosis.

METHODSThe total RNA from human liver tissue of cirrhosis was extracted using Trizol method, and the mRNA was purified using mRNA purification kit. SMART technique and CDSIII/3' primer were used for first-strand cDNA synthesis. Long distance PCR was then used to synthesize the double-strand cDNA that was then digested by proteinase K and Sfi I, and was fractionated by CHOMA SPIN-400 column. The cDNA fragments longer than 0.4 kb were collected and ligated to lambdaTripl Ex2 vector. Then lambda-phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries was strictly checked by conventional titer determination. Eleven plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector.

RESULTSThe titers of unamplifed and amplified libraries were 1.03 x 10(6) pfu/ml and 1.36 x 10(9) pfu/ml respectively. The percentages of recombinants from both libraries were 97.24 % in unamplified library and 99.02 % in amplified library. The lengths of the inserts were 1.02 kb in average (36.36 % 1 approximately equals 2 kb and 63.64 % 0.5 approximately equals 1.0 kb).

CONCLUSIONA high quality cDNA library from human liver tissue of cirrhosis was constructed successfully, which can be used for screening and cloning new special genes associated with the occurrence of cirrhosis.

DNA, Complementary ; biosynthesis ; Gene Library ; Hepatitis B, Chronic ; complications ; genetics ; virology ; Humans ; Liver Cirrhosis ; genetics ; virology ; Polymerase Chain Reaction ; Recombination, Genetic ; Transcription, Genetic ; genetics

DNA, Viral ; analysis ; Hepatitis B Core Antigens ; analysis ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Immunohistochemistry ; Liver ; virology ; Liver Cirrhosis ; virology ; Oligonucleotide Array Sequence Analysis ; methods

OBJECTIVETo investigate the distribution of hepatitis B virus (HBV) genotype in Guizhou and to study the relationship between the genotype and the progression of liver disease.

METHODS786 patients with chronic HBV infection, from 4 cities of Guizhou, including 346 asymptomatic carriers (ASC), 313 chronic hepatitis (CH), 77 liver cirrhosis (LC), 50 hepatocellular carcinoma (HCC) were examined. HBV genotype was determined by restriction fragment length polymorphism analysis and the subtypes were determined by direct sequencing of PCR product in 94 patients with HBV B genotype, the relationship between HBV genotype and the progression of liver disease was studied by multifactor analysis such as HBeAg positivity, HBV DNA load and ALT level.

RESULTSOf the 786 patients, 7 (0.89%), 497 (63.23%), 275 (34.99%), and 7 (0.89%) belonged to genotype A, B, C, D, respectively. There was statistically significant difference in the distribution of genotype B among Kaili (96.04%), Zunyi (78.79%), Duyun (64.52%) and Guiyang (53.14%) (P< 0.01). Genotype C was more prevalent in Guiyang than in other three cities (P < 0.01, or P < 0.05). Out of 94 genotypes B, 93 (98.94%) belonged to subtype Ba, only one was subtype Bj. There were statistically significant difference in the distribution of genotype B and C among various stage of liver disease (P < 0.05 or P < 0.01). Genotype B showed a gradual decrease from ASC, CH, LC to the HCC group while in contrast, genotype C showed a gradual increase in the same order. The ALT levels and the mean age were significantly higher and older in patients with genotype C than those in genotype B (P < 0.01 or 0.05). The HBeAg positivity was significantly lower in genotype C than that in genotype B (P < 0.025).

CONCLUSIONData showed that there were genotype A, B, C and D existing in Guizhou. Genotype B was the major one but genotype C was more commonly seen. In genotype B, subtype Ba appeared to be predominant. The geographic distribution of genotype B and C were different in some cities of Guizhou. Compared to genotype B, genotype C was associated with the development of more severe liver damage.

Carcinoma, Hepatocellular ; pathology ; virology ; DNA, Viral ; analysis ; Disease Progression ; Genotype ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; genetics ; pathology ; Humans ; Liver ; pathology ; Liver Cirrhosis ; pathology ; virology ; Liver Neoplasms ; pathology ; virology ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length