Background: Human Pegivirus (HPgV), previously called Hepatitis G virus or GB virus C, is an RNA virus. It can be transmitted vertically (mother to infant), parenterally and sexually. HPgV share common routes of transmission to other viruses such as Hepatitis B virus, Hepatitis C virus and Human Immunodeficiency virus (HIV) thus co-infection is usually observed. Risk groups of HPgV include injection drug users, HIV-positive individuals, multi-transfused patients, hemodialysis patients, hemophiliacs, chronic liver disease patients and organ transplant recipients. The clinical significance of HPgV is not yet established and warrants further studies. Research on HPgV in the Philippines is scarce and has not been updated for over 10 years. There is no published data on HPgV prevalence in Filipino pediatric population specifically among risk groups like multi-transfused children with decompensated liver disease secondary to biliary cirrhosis and liver transplant pediatric patients. The lack of local data warrants conduct of this study. Objective: To determine the presence of HPgV RNA, HPgV E2 antibody (anti-E2) and HBsAg among Filipino children with decompensated liver disease secondary to biliary cirrhosis (DBC) and liver transplant pediatric patients (LTP). Methods: Included were 15 children with DBC and 15 LTP recruited from the Section of Pediatric Gastroenterology, Hepatology and Nutrition of the UP PGH. All patients’ sera were tested for HPgV RNA by Real Time RT-PCR, HPgV anti-E2 by Enzyme-linked Immunosorbent Assay (ELISA) and hepatitis B surface antigen (HBsAg) by immunochromatographic test. Twenty age and sex matched children with no history of liver disease and blood transfusion served as controls. Results: All patient and control samples were negative for HPgV RNA. HPgV anti-E2 was detected in 6 of 15 LTP, 5 of 15 DBC and 1 of 20 controls. HBsAg was detected in 2 of 15 LTP, 5 of 15 DBC and 0 of 20 controls. Four patients (two LTP, two DBC) were positive for both HPgV anti-E2 and HBsAg. Conclusion This study showed that a proportion of liver transplant patients and those with decompensated biliary cirrhosis are positive for HPgV anti-E2, which indicates that these individuals previously had HPgV infection but is now resolved. Possible source of infection is infected blood from the blood transfusions, infected transplant organ or infected mother. Since routine HPgV screening is not yet recommended for the general population, blood donors and organ donors, the confirmation of exact source of infection may be difficult. Co-infection with HBsAg was also observed in both risk groups which suggests that at some point in time, these children were infected by both HPgV and HBV and also the possibility of simultaneous infection by the two viruses. This study provides preliminary data on the proportion of HPgV infection in Filipino children belonging to two of the HPgV risk groups. Studies with a larger and more significant sample size to determine HPgV prevalence as well as studies regarding the pathogenicity of HPgV are warranted. As this may provide basis for routine HPgV screening among risk groups and blood donations in the future.
GB virus C

Adult ; China ; epidemiology ; GB virus C ; classification ; Hepatitis, Viral, Human ; epidemiology ; virology ; Homosexuality, Male ; Humans ; Male

Several research groups have recently reported that persistent GB virus C (GBV-C) co-infected with human immunodeficiency virus (HIV) leads to slower AIDSs disease progression than HIV-1 infection alone. However, these findings were not confirmed by several other studies. To investigate the association between GBV-C replication and plasma HIV loads and CD4+ T cell counts, 203 HIV-1 positive former blood/plasma donors(FBDs) were enrolled from Fuyang city of Anhui Province in China. Plasma specimens were collected from them and were tested for GBV-C using RT-PCR and ELISA. Out of 203 specimens, 52 (25.6%) cases were positive for GBV-C, including 35 male (67.3%) and 17 female (32.7%) cases. No significant association was identified between GBV-C infection and CD4+ T-cell counts or between GBV-C infection and HIV viral loads. Since all the subjects studied were naive to ART, the influence of therapy on AIDS disease progression was ruled out in this study. Overall, our data indicated that HIV-1 positive male FBDs were prone to be infected, GBV-C coinfection with HIV-1 does not significantly influence HIV/AIDS disease progression during the late stage of chronic HIV-1 infection.
Acquired Immunodeficiency Syndrome ; complications ; immunology ; virology ; Adult ; Aged ; CD4 Lymphocyte Count ; Disease Progression ; Female ; Flaviviridae Infections ; immunology ; virology ; GB virus C ; HIV-1 ; physiology ; Hepatitis, Viral, Human ; immunology ; virology ; Humans ; Male ; Middle Aged ; RNA, Viral ; blood ; 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

OBJECTIVETo study the simple infection and super/co-infection of HAV-HEV, HGV in patients with viral hepatitis.

METHODSUsing EIA method to detect anti-HAV IgM, HBV serum markers, anti-HCV IgM, anti-HDV IgM, anti-HEV IgM, anti-HGV IgM in viral hepatitis patients with different clinical types.

RESULTSSeventy-three percent patients (154/210) had HBV infection markers, twenty-nine percent patients (61/210) had HAV infection marker, eight percent patients (17/210) had HCV, HDV infection markers, ten percent patients (21/210) had HEV infection and seven percent patients (15/210) had HGV infection. Only nine percent patients (20/210) had viral hepatitis serum markers negative. In all clinical types, sixty-one percent patients had only one type hepatitis virus infection, thirty-two percent patients had two types of hepatitis virus super/co-infection, six percent patients had three types of hepatitis virus super/co-infection. Super/co-infection often occurred in patients who had cirrhosis or hepatic failure.

CONCLUSIONHBV and HAV infection is very common in viral hepatitis patients, whereas HCV, HDV, HEV and HGV infection is relatively low; double super/co-infection of HAV-HEV, HGV frequently occurs in severe patients with viral hepatitis.

Antibodies, Viral ; blood ; China ; epidemiology ; Female ; GB virus C ; isolation & purification ; Hepatitis A ; epidemiology ; virology ; Hepatitis A virus ; isolation & purification ; Hepatitis E ; epidemiology ; virology ; Hepatitis E virus ; isolation & purification ; Hepatitis Viruses ; isolation & purification ; Hepatitis, Viral, Human ; epidemiology ; virology ; Humans ; Male ; Superinfection

In order to study the feasibility of E2 gene fragment of hepatitis virus G(HGV) as a component of DNA vaccine against the hepatitis virus G infection, a 559bp DNA fragment encoding HGV E2 was cloned into plasmid pCMV-S from pThioHis-E2 in the same reading frame with HBsAg gene to form a recombinant plasmid named pCMV-S-E2. BALB/c mice of Kunming strain were immunized with purified plasmid DNA of pCMV-S-E2 by intra-muscularly inoculation. The immunizations were boosted twice at an interval of 14 days. The whole blood was collected from mice orbit on the day-8 after the last boost. Mice sera were screened by ELISA to determine the humoral immune response using E2-GST fusion protein as the immobilized antigen and the sera from mice immunized with pCMV-S as control. The result indicated that the immunization with plasmid DNA of pCMV-S-E2 could induce quite strong humoral immune response.
Animals ; Female ; GB virus C ; immunology ; Hepatitis Antibodies ; blood ; Mice ; Mice, Inbred BALB C ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; immunology ; Vaccines, DNA ; immunology ; Viral Envelope Proteins ; genetics ; immunology ; Viral Hepatitis Vaccines ; immunology

OBJECTIVETo realize human immunodeficiency virus(HIV) and hepatitis C virus(HCV) super-infected with hepatitis G virus(HGV or GBV/C) and to probe into the mechanism of these virus infection in the body.

METHODSHIV and HCV load were tested by the quantitated RT-PCR in the HIV or HCV infected plasma samples respectively and the HGV RNA was detected in all of the samples. Then some of the HGV positive were sequenced.

RESULTS123 of 317 HIV patients were positive for HGV, the positive rate was 38.8%. Among the 91 HCV patients, 19 were positive for HGV. The positive rate is 20.9% which was less than that of HIV patients. HIV load of the patients super-infected with HGV was less than that of those without HGV[(1.8+/-0.6)x10 copies/ml compared with (1.9+/-1.1)x10(2)copies/ml]; while HGV and HCV super-infection did not influence the HCV RNA load significantly [(1.5+/-0.6)x10(4) copies/ml compared with (5.4+/-1.8)x10(4)copies/ml]. The HGV sequences from HIV or HCV patients were compared and showed no difference markedly.

CONCLUSIONThe rate of the HIV and HGV super-infection is higher than that of HCV. HGV may inhibit HIV reproduction in the body while superinfection.

GB virus C ; HIV ; physiology ; HIV Infections ; virology ; Hepacivirus ; physiology ; Hepatitis C ; virology ; Hepatitis, Viral, Human ; virology ; Humans ; RNA, Viral ; blood ; Virus Replication

OBJECTIVETo observe the serological features of hepatitis G virus (HGV or GBV) in selected population of Shaanxi, Qinghai and Xinjiang Provinces (region).

METHODSEnzyme-linked immunosorbent assay (ELISA) was used to determine IgG antibody against GBV (anti-GBV IgG) in serum specimens of 1469 individuals from the 3 provinces (region).

RESULTSThe positivity rate of serum GBV-IgG antibody in ethnic minorities (4.11% in Tibetan, 5.36% in Mongolian, 4.55% in Uigur, 4.00% in Hui population) was slightly higher than that in Han population (1.36%-1.73%), but the differences were not significant (P>0.05). The positivity rate of serum GBV-IgG antibody in drug abusers (11.30%, 34/301) was remarkably higher than that of the normal population (2.44%,18/736) (P<0.01). The positivity rate of serum GBV-IgG antibody in blood donors was 1.02%-7.68%.

CONCLUSIONSThe positivity rates of serum GBV-IgG antibody among ethnic groups in the 3 provinces (region) had no significant differences; blood-borne transmission seemed to be an important transmission route of GBV, therefore supervision of blood donors and drug abusers should be intensified.

Antibodies, Viral ; blood ; Blood Donors ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Flaviviridae Infections ; epidemiology ; transmission ; GB virus C ; immunology ; Hepatitis, Viral, Human ; epidemiology ; transmission ; Humans ; Immunoglobulin G ; blood ; Seroepidemiologic Studies ; Substance-Related Disorders ; virology

OBJECTIVETo observe the relationship between the amount of HBV DNA in serum/liver tissue and HGV infection in patients with chronic hepatitis B (CH-B) for exploring the effect of HGV infection on hepatitis B virus (HBV) replication of CH-B.

METHODSHGV RNA in serum, HGV nonstructural region 5 (NS5) antigen (HGV Ag) in liver tissue and the amount of HBV DNA in serum, liver tissue were detected for 56 patients with CH-B by reverse transcription-polymerase chain reaction (RT-PCR) assay, peroxidase antiperoxidase (PAP) immunohistochemical method and fluorescence quantitative PCR assay, respectively. Then the relationship between HGV Ag expression in liver tissue and HGV RNA expression in serum was analysed and the amount of HBV DNA in serum and liver tissues from the serum HGV RNA or liver tissue HGV Ag positive patients were compared with those of the serum HGV-RNA or liver tissue HGV Ag negative patients, respectively.

RESULTSTen (17.9%) and eight (14.3%) patients were positive for serum and liver tissues,respectively.HGV RNA expression in serum was closely related to HGV Ag expression in liver tissues, but there was HGV RNA in serum from some of the liver tissues HGV Ag negative patients ?cases of HGV RNA and HGV Ag positive or negative,HGV RNA positive but HGV Ag negative, HGV RNA negative but HGV Ag positive, respectively: 5,43,5,3,(P<0.01). There was no significant difference in the amount of HBV DNA in serum and liver tissues between HGV RNA or HGV Ag positive and negative patients (P>0.05).

CONCLUSIONSHGV infection may not affect HBV replication. Liver is the site of HGV replication, but HGV probably also replicates in extrahepatic tissues. HGV hepatic pathogenicity is probably mild and further studies are still needed.

Adult ; DNA, Viral ; analysis ; blood ; Female ; Flaviviridae Infections ; complications ; virology ; GB virus C ; genetics ; immunology ; pathogenicity ; Hepatitis Antigens ; analysis ; Hepatitis B virus ; genetics ; physiology ; Hepatitis B, Chronic ; complications ; virology ; Hepatitis, Viral, Human ; virology ; Humans ; Liver ; virology ; Male ; RNA, Viral ; blood ; Virus Replication

A cDNA fragment locating at the putative envelop protein 2(E2) region of GBV-C/HGV fused with Schistosoma japonicum, glutathione S-transferase(GST) was amplified with PCR from plasmid pGEX-E2. The amplified DNA fragment was inserted into plasmid pGEX-5X-1, at the downstream of the coding sequences of GST, in the same reading frame with the gene of GST. The fusion gene fragment of GST-E2 was amplified with PCR, using the recombinant plasmid pGEX-5X-1-E2 as the template. The amplified 1324 bp DNA fragment of GST-E2 was inserted into Pichia pastoris expression vector pPIC9K in reading frame with alpha-factor secreting signal peptide. The plasmid pPIC9K-GST-E2 was transformed into Pichia pastoris GS115 with electroporation. The transformants (His+ Muts) were selected and induced to express the 54kD GST-E2 fusion protein, which could be specially recognized by both the antisera directed against E2 and against GST. The GST-E2 fusion protein was purified with Sepharose 4B glutathione affinity chromatography to a purity of 95%. The expression was optimized to achieve the highest expression level of GST-E2 fusion protein which was accumulated up to 50% of total proteins in the culture supernatant. The GST-E2 protein derived from the recombinant Pichia pastoris was proved possessing antigenicity and high specificity by ELISA, probed with sera from the patients infected by GBV-C/HGV.
Animals ; Antigens, Viral ; genetics ; immunology ; isolation & purification ; GB virus C ; genetics ; immunology ; Gene Expression ; Genetic Engineering ; Glutathione Transferase ; genetics ; Hepatitis Antibodies ; blood ; immunology ; Humans ; Pichia ; Recombinant Fusion Proteins ; genetics ; immunology ; isolation & purification ; Schistosoma japonicum ; enzymology ; Viral Envelope Proteins ; genetics ; immunology ; isolation & purification