To evaluate the clinical feasibility of the antibody titer against a chimeric polypeptide (named Core 518), in which a domain of Core and NS3 of hepatitis C virus (HCV) was fused, ELISA was performed in a total of 76 serum samples. Each serum was serially diluted using two-fold dilution method with distilled water into 10 concentrations. They were all positive for second generation anti-HCV assay (HCV EIA II; Abbott Laboratories). Genotyping RT-PCR, quantitative competitive RT-PCR, and RIBA (Lucky Confirm; LG Biotech) were also assayed. Anti-Core 518 antibody was detected in x 12800 or higher dilutions of sera from 35 of 43 chronic hepatitis C (81.4%) and nine of 16 hepatocellular carcinoma sera (56.3%), one of four cirrhosis (25%), 0 of four acute hepatitis C, and one of nine healthy isolated anti-HCV-positive subjects (p=0.0000). The anti-Core 518 antibody titers were well correlated with the presence of HCV RNA in serum (p=0.002). The anti-Core 518 antibody titers decreased significantly in nine of ten responders to IFN-alpha treatment. Monitoring anti-Core 518 titers may be helpful not only for differentiating the status of HCV infection among patients with various type C viral liver diseases, but also for predicting responses to IFN-alpha treatment.
Adult ; Aged ; Female ; Genotype ; Hepatitis C/immunology* ; Hepatitis C/drug therapy* ; Hepatitis C/diagnosis ; Hepatitis C/blood ; Hepatitis C Antibodies/immunology* ; Hepatitis C Antibodies/blood ; Hepatitis C Antigens/immunology* ; Hepatitis C-Like Viruses/immunology* ; Hepatitis C-Like Viruses/genetics ; Human ; Immunoblotting ; Interferon Alfa-2a/therapeutic use* ; Male ; Middle Age ; RNA, Viral/blood ; Recombinant Fusion Proteins/immunology ; Viral Core Proteins/immunology* ; Viral Nonstructural Proteins/immunology*

Hepacivirus

The incidence and clinical significance of GB virus C/hepatitis G virus(GBV-C/HGV) infection were evaluated in 68 patients on maintenance hemodialysis. GBV-C/HGV RNA was identified in serum by a reverse transcription-polymerase chain reaction assay with nested primers deduced from a nonstructural region. Hepatitis B surface antigen(by RIA) and anti-hepatitis C(by ELISA) were checked simultaneously. Out of 68 patients, GBV-C/HGV RNA was detected in 5(7.4%), HBsAg in 4 patients(5.9%) and anti-HCV in 15 patients(22%). All 5 patients with positive GBV-C/HGV RNA had a history of blood transfusion. Out of 5 patients with positive GBV-C/HGV RNA, 2 patients were coinfected with hepatitis C virus, who showed chronic hepatitis clinically. Three patients with isolated GBV-C/HGV infection showed normal liver function during last 18 months' period. Between the patients with positive GBV-C/HGV RNA and those with negative GBV- C/HGV RNA, there was no difference in age, sex, duration of hemodialysis and amount of transfusion. Our data suggest that GBV-C/HGV infection may be present, with or without hepatitis C virus infection, in maintenance hemodialysis patients. Although the liver function of patients with isolated GBV-C/ HGV infection was normal, the clinical significance of this new virus remains to be determined.
Blood Transfusion ; GB virus C ; Hepacivirus ; Hepatitis ; Hepatitis B ; Hepatitis B Surface Antigens ; Hepatitis, Chronic ; Humans ; Incidence* ; Liver ; Renal Dialysis* ; RNA

Genetic changes between codons 2209 and 2248 of NS5A of genotype 1b hepatitis C virus (HCV-1b) have been reported to be associated with the sensitivity to interferon-alpha (IFN-alpha). The present study was performed to analyze such relationship in Korean patients with chronic hepatitis C and HCV-1b (n=19), including 12 chronic hepatitis C patients treated with IFN-alpha, 3 chronic hepatitis C patients without treatment as controls, and 4 patients with hepatocellular carcinoma (HCC). Two serum samples, before and after the treatment, were analyzed for the mutations by reverse transcription-polymerase chain reaction, cloning and sequencing. The mutations were identified in 32% (6/19), including five intermediate type (1-3 mutations) and one mutant type (4 or more). In 12 patients treated with IFN-alpha, the number of amino acid substitutions in NS5A2209-2248 was not associated with outcome of the treatment.
Adult ; Aged ; Amino Acid Sequence ; Antiviral Agents/therapeutic use* ; Base Sequence ; Carcinoma, Hepatocellular/virology* ; Carcinoma, Hepatocellular/blood ; Codon ; Female ; Genotype ; Hepatitis C, Chronic/virology* ; Hepatitis C, Chronic/drug therapy* ; Hepatitis C, Chronic/blood ; Hepatitis C-Like Viruses/isolation & purification ; Hepatitis C-Like Viruses/genetics* ; Hepatitis C-Like Viruses/classification ; Human ; Interferon-alpha/therapeutic use* ; Liver Neoplasms/virology* ; Liver Neoplasms/blood ; Male ; Middle Age ; Molecular Sequence Data ; Mutation* ; Prognosis ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND: Although the polyproteins of hepatitis C virus(HCV) are processed and formed in nearly equimolar amounts, individual functional proteins have a discrepancy in their time of appearance following HCV infection and eliciting immune response. This study was conducted to compare the reactivity toward regional specific HCV protein in relation to virological characteristics, including HCV genotype and HCV replication. METHODS: Sera from forty-five patients with chronic HCV infection were analyzed through the experiments of the recombinant immunoblot assay(RIBA-2), HCV genotyping and HCV RNA quantitation. RESULTS: The frequencies of seropositivity to C22-3, C33C, C100-3 and 5-1-1 proteins were 91.1+ACU-, 91.1+ACU-, 64.4+ACU- and 53.3+ACU-, respectively, of all the patients, and thus the antibodies to C22-3 and C33C proteins were found more frequently (p +ADw- 0.05). The antibody responses between core or NS3 proteins and NS4 proteins showed more discrepancy in the HCC group than that in the CH group, implying a possibility of oncogenic potential of core or NS3 gene in hepatocarcinogenesis. The detection rate of antibodies to C22-3 and C33C, in accordance with serum HCV RNA levels, was significantly higher in highly viremic patients than that in low viremic patients (p +ADw- 0.05). Antibodies to C22-3, C33C, C100-3 and 5-1-1 were also found more frequently in patients with HCV genotype 1b, compared to those with HCV genotype 2a (p +ADw- 0.05). CONCLUSION: These results suggest that antibody detection of HCV may depend on the virological characteristics of HCV, the levels of HCV replication and HCV genotype and, therefore, HCV RNA detection using RT-PCR technique is essential for confirmatory diagnosis for HCV infection. Furthermore, the HCV core or NS3 Protein may play important role in hepatocarcinogenesis.
Adult ; Aged ; Female ; Genotype ; Hepatitis C Antibodies/blood+ACo- ; Hepatitis C, Chronic/virology ; Hepatitis C, Chronic/immunology+ACo- ; Hepatitis C-Like Viruses/physiology ; Hepatitis C-Like Viruses/genetics ; Human ; Male ; Middle Age ; RNA, Viral/blood ; Viral Core Proteins/immunology+ACo- ; Viral Nonstructural Proteins/immunology+ACo- ; Virus Replication

Infectious agents, including viruses, bacteria and parasites, can be transmitted via human blood and blood products. Of greatest importance are viruses such as human immunodeficiency virus types 1 and 2 (HIV-1/2), hepatitis B virus (HBV), and hepatitis C virus (HCV), followed by other viruses such as cytomegalovirus (CMV) and human parvovirus B19. Viruses such as hepatitis G virus and TT virus can also be transmitted via blood products, but their pathogenicity is still unclear. Bacteria, including Treponema pallidum and Yersinia enterocolitica and parasites such as Plasmodium species can also be transmitted from donors to recipients. Furthermore, the threat of newly emerging pathogens that can affect the blood safety, such as the variant Creutzfeld-Jakob Disease, is always present. The measures to reduce the risks of transfusiontransmitted infection within the last 20 years, such as donor selection and testing donated blood for various infectious agents, have had a remarkable impact on the safety of blood supply. Nevertheless, the public expectation of absolute blood safety continues to exert pressure to eliminate the remaining risks. The recent introduction of molecular biology techniques combined with viral inactivation methods is directed to get this goal.
Bacteria ; Blood Safety ; Cytomegalovirus ; Donor Selection ; GB virus C ; Hepacivirus ; Hepatitis B virus ; HIV ; Humans ; Molecular Biology ; Parasites ; Parvovirus B19, Human ; Plasmodium ; Tissue Donors ; Torque teno virus ; Treponema pallidum ; Virulence ; Virus Inactivation ; Yersinia enterocolitica

BACKGROUND: A recently identified Flaviviridae-like agent, termed hepatitis G virus (HGV), has been recognized as a non A-E hepatitis agent, but its relation to liver disease and transmission mode are not well understood. We investigated HGV infection rate in Korea and tried to clarify its relation to the liver disease. METHODS: 145 blood donors, 39 hemodialysis patients and 22 hepatitis C virus (HCV) infected persons were investigated for the presence of HGV by nested reverse transcriptase polymerase chain reaction (nested RT-PCR) with primers from the 5' UTR of HGV and some liver function tests. In each PCR assay, one positive and two negative controls were included. RESULTS: HGV-RNA was detected in 11 (7.6%) of 145 young voluntary blood donors and in 5 (12.8%) of 39 hemodialysis patients and in 8 (36.4%) of 22 HCV infected patients. All HGV RNA positive hemodialysis patients have a past history of transfusion, but they had a remarkably shorter duration of hemodialysis than those of HGV-negative patients. HCV infected patients with HGV-RNA tended to be younger than those without HGV-RNA. In all 15 HGV-RNA infected individuals without hepatitis B and C infection, alanine amino transferase was not increased except in 2 cases. Liver function tests did not show a significant difference between HGV-RNA positive patients and negative patients. CONCLUSIONS: Hepatitis G virus infection rate was much higher in Korea than other countries, so we suggested that group life could be another transmission mode other than blood transfusion. But even in infected cases, HGV did not seem to cause hepatitis and a high proportion cleared the virus after a relatively short time.
5' Untranslated Regions ; Alanine ; Blood Donors* ; Blood Transfusion ; GB virus C* ; Hepacivirus ; Hepatitis B ; Hepatitis B virus ; Hepatitis C* ; Hepatitis* ; Humans ; Korea ; Liver Diseases ; Liver Function Tests ; Polymerase Chain Reaction ; Renal Dialysis* ; Reverse Transcriptase Polymerase Chain Reaction ; RNA ; Transferases

BACKGROUND: Transforming growth factor beta-1 (TGF beta 1) has been suggested to play a role in the development, growth or progression of hepatocellular carcinoma (HCC). Genotype and serum titer of HCV also affect the occurrence of HCC in chronic hepatitis C. In this study, we were to evaluate the effects of genotype or serum titer of HCV on the expression of TGF beta 1. We also intended to examine the correlation between the up-regulation of TGF beta 1 and the association with HCC in patients with chronic hepatitis C. METHODS: We studied 19 patients with chronic hepatitis C and 18 with HCC associated with HCV infection. HCV genotype was determined by line probe reverse hybridization assay and the amount of HCV-RNA was quantitated by branched DNA signal amplification assay. Serum TGF beta 1 level was measured by enzyme linked immunosorbent assay. RESULTS: HCV genotypes of patients with HCC were similar to those without it. Serum HCV-RNA titer was higher in genotype 1b than in non-1b (p < 0.05). Serum TGF beta 1 levels were higher in HCC than in chronic hepatitis (p < 0.05). However, there was no significant difference in the serum TGF beta 1 level between genotype 1b and non-1b. Also, it was not correlated with the serum HCV-RNA titer or alanine aminotransferase levels. CONCLUSION: TGF beta 1 seems to be overexpressed in HCC compared to that of chronic hepatitis C: it was not affected by serum ALT levels, genotype or serum HCV titer. It is suggested that TGF beta 1 may be associated with the malignant transformation of hepatocyte or the progression of HCV-associated HCC.
Adult ; Aged ; Alanine Transaminase/blood ; Carcinoma, Hepatocellular/virology ; Carcinoma, Hepatocellular/metabolism* ; Female ; Genotype ; Hepatitis C, Chronic/virology ; Hepatitis C, Chronic/metabolism* ; Hepatitis C-Like Viruses/genetics ; Hepatitis C-Like Viruses/classification ; Human ; Liver Neoplasms/virology ; Liver Neoplasms/metabolism* ; Male ; Middle Age ; RNA, Viral/blood ; Transforming Growth Factor beta/blood*

The positive predictability of anti-HCV ELISA is low, especially, in blood donors and in healthy populations. False positive anti-HCV results pose some difficulties in medical practice and in blood screening. The aim of this study was to identify the factors associated with true hepatitis C virus (HCV) infection among anti-HCV ELISA-positives. A case-control analysis was conducted using 354 subjects who were positive for anti-HCV ELISA. All subjects were tested for true HCV infection using the reverse transcriptase polymerase chain reaction (RT-PCR). Tests for serum alanine aminotransferase (ALT), fasting glucose, HBsAg, anti-HBc antibody, alpha-fetoprotein, platelet count and ultrasound of liver were also performed. Epidemiological data were obtained by self-administered questionnaires. Out of 354 subjects, 202 (57.1%) were positive for HCV by RT-PCR and 152 were negative and used as the control group. In multivariate analysis, blood transfusion (odds ratio, OR 2.3, 95% confidence interval, CI 1.3-4.0), elevated ALT (OR 2.2, 95% CI 1.2-4.3) and higher anti-HCV ELISA ratios (more than 3; OR 1.7, 95% CI 1.3-2.1) were associated with true HCV infection. Thrombocytopenia was also associated with the presence of HCV in univariate analysis. These results suggest that a history of blood transfusion, elevated ALT and a high score on anti-HCV ELISA ratios are associated with true HCV infection among anti-HCV ELISA-positives.
Adult ; Blood Donors ; Enzyme-Linked Immunosorbent Assay* ; False Positive Reactions ; Female ; Hepatitis C/epidemiology ; Hepatitis C/diagnosis* ; Hepatitis C/blood ; Hepatitis C Antibodies/blood* ; Hepatitis C-Like Viruses ; Human ; Male ; Middle Age ; Multivariate Analysis ; Predictive Value of Tests ; Reverse Transcriptase Polymerase Chain Reaction ; Risk Factors

To determine the prevalence and clinical relevance of HGV infection in dialysis patients, we performed a cross-sectional study of 61 HD patients and 79 Continuous Ambulatory Peritoneal Dialysis (CAPD) patients. HGV-RNA was identified by reverse-transcription (RT) polymerase chain reaction (PCR) assay with primers from the 5'-untranslated region of the viral genome. The prevalence of HGV infection was similar in HD and CAPD patients (9.8% vs. 12.7%), while that of HCV infection was significantly higher in HD patients compared to CAPD patients (16.4% vs. 1.3%, p < 0.05). The mean age (49.2 +/- 13.4 vs. 46.7 +/- 13.0 years), male to female ratio (2.4:1 vs. 1.3:1), history of transfusion (62.3% vs. 49.4%), history of hepatitis (27.9% vs. 26.6%), mean ALT level during the previous 6 months (22.4 +/- 37.9 vs. 14.0 +/- 7.4 IU/L), and the prevalence of HBsAg (8.2% vs. 6.3%) showed no difference between HD and CAPD patients. In both HD and CAPD patients, the presence of HGV RNA was not related to age, sex, duration of dialysis, history of transfusion, history of hepatitis, or to the presence of HBV or HCV markers. There was no significant difference in the clinical and biochemical data between patients with isolated HGV infection (n = 12) and patients without viremia (n = 106). The clinical feature of patients coinfected with HGV and HBV (n = 2), or HGV and HCV (n = 2) seemed to be similar to those of patients with isolated HBV (n = 8) or HCV (n = 9) infection. In conclusion, the prevalence of HGV infection was not different between HD and CAPD patients, and HGV infections did not seem to be associated with clinically significant hepatitis. The routes of HGV transmission, other than transfusion or contamination during HD procedure, were suspected.
Female ; Hepatitis Agents, GB*/genetics ; Hepatitis C/genetics ; Hepatitis C-Like Viruses/genetics ; Hepatitis, Viral, Human/virology ; Hepatitis, Viral, Human/genetics ; Hepatitis, Viral, Human/etiology* ; Human ; Male ; Middle Age ; Peritoneal Dialysis, Continuous Ambulatory/adverse effects* ; Prevalence ; RNA, Viral/analysis ; Renal Dialysis/adverse effects* ; Viremia/genetics