Animals ; China ; epidemiology ; Disease Reservoirs ; Hepatitis E ; epidemiology ; transmission ; Hepatitis E virus ; genetics ; isolation & purification ; Humans ; Rodentia ; virology ; Swine ; virology

Hepatitis E, caused by hepatitis E virus (HEV) infection, usually leads to an acute clinical course, and is the most common diagnosis among cases of acute viral hepatitis. From 2008, there have been increasing reports of chronic HEV infection in immunocompromised patients such as organ transplant recipients. Without intervention with antiviral treatment, approximately 60% of HEV infections in organ transplant recipients evolve into chronic HEV infections. Of these chronic hepatitis E patients, 10% may develop liver fibrosis and progress to liver cirrhosis. This article reviews chronic HEV infection and treatment in organ transplant recipients.
Animals ; Antiviral Agents ; therapeutic use ; Hepatitis E ; drug therapy ; virology ; Hepatitis E virus ; genetics ; isolation & purification ; physiology ; Hepatitis, Chronic ; drug therapy ; virology ; Humans ; Transplant Recipients ; Transplants ; virology

OBJECTIVETo investigate the presentation of a neutralization epitope-containing peptide antigen of hepatitis E virus (HEV) on chimeric virus-like particles (VLPs) of hepatitis B surface antigen (HBsAg).

METHODSThe gene fragment corresponding to amino acids (aa) 551-607 (HEnAg) of HEV capsid protein, which contains the only neutralization epitope identified to date, was fused via a synthetic glycine linker in frame with the gene of HBsAg. The resulted fusion gene was then integrated through transformation into the genome of Pichia pastoris under the control of a methanol-induced alcohol oxidase 1 (AOX1) promoter and expressed intracellularly. The expression products in the soluble cell extracts were characterized by Western blot, ELISA, CsCl density gradient analysis, and electron microscopic visualization.

RESULTSThe novel fusion protein incorporating HBsAg and the neutralization epitope-containing HEnAg was expressed successfully in Pichia pastoris with an expected molecular weight of approximately 32 kD. It was found to possess the ability to assemble into chimeric HBV/HEV VLPs with immunological physical and morphological characteristics akin to HBsAg particles. Not only did the chimeric VLPs show high activity levels in a HBsAg particle-specific ELISA but they were also strongly immunoreactive with hepatitis E (HE) positive human serum in a HEV specific ELISA, indicating that HEnAg peptide fragments were exposed on VLP surfaces and would be expected to be readily accessible by cells and molecules of the immune system. Similarity between chimeric VLPs to highly immunogenic HBsAg particles may confer good immunogenicity on surface-displayed HEnAg.

CONCLUSIONThe chimeric HBV/HEV VLPs produced in this study may have potential to be a recombinant HBV/HEV bivalent vaccine candidate.

Epitopes ; Genetic Engineering ; Hepatitis Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis E virus ; genetics ; immunology ; Pichia ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Vaccines, Synthetic

BACKGROUNDTo establish a new assay for detecting the quantity of HBV DNA with PCR and enzyme-linked immunosorbent assay(ELISA).

METHODSThe products of PCR using primers pre-labeled with biotin were hybridized with the capture probes that were immobilized on the microtiter strips and then bond with Sav-Ap. The quantity of DNA was detected by measuring the yellow color at 405 nm wave length.

RESULTSTotally 125 sera from patients with hepatitis B were tested for HBV DNA by this method,the sera were also tested for HBV immunological markers by solid phase radio immuno-assay (SPRIA). The HBV positive rate with PCR-ELISA was 64.9% (24/37) in samples which were positive for HBsAg, HBeAg and HBcAb; and 34.2% (13/38) in sera which were positive for HBsAg, HBeAb and HBcAb; in sera positive for HBsAg and HBcAb or only HBcAb, the positive rate was 6.7% (1/15) and 5.9% (2/34) respectively.

CONCLUSIONSThe PCR ELISA assay is simple and suitable for clinical laboratory in quantitative determination of HBV DNA.

DNA Probes ; DNA, Viral ; analysis ; biosynthesis ; genetics ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Polymerase Chain Reaction ; Sensitivity and Specificity

OBJECTIVETo explore the relationship between HBV core promoter mutations and liver damage or HBeAg status.

METHODSNested polymerase chain reaction (nPCR) was used for amplification of HBV DNA core promoter in 59 sera from patients with chronic hepatitis B in Guangxi, then the HBV DNA positive products were sequenced by direct sequencing.

RESULTSThe HBV DNA positive rate of was 59.3%(35/59). All the patients were infected by mutants. The commonest mutation was the double mutation (A --> T at nt1762 and G --> A at nt1764), counting for 57.1% (20/35). The next was C --> G at nt1799, counting for 54.4% (19/35), but this was no function. A --> G at nt1752 (resulting in isoleucine to valine) was seen in 37.1% (13/35) of the HBV DNA positive patients, and T --> C at nt1753 was seen in 20% (7/35). The significant difference in the frequency of T1762A1764 mutant was found between HBeAg positive patients (31.3%) and negative patients (79.0%).

CONCLUSIONSHBV core promoter mutations are common among patients with chronic hepatitis B in Guangxi. T1762A1764 mutant is associated with HBeAg status and chronic hepatitis.

Adolescent ; Adult ; Female ; Hepatitis B Core Antigens ; genetics ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis B, Chronic ; virology ; Humans ; Liver ; pathology ; Male ; Middle Aged ; Point Mutation ; Promoter Regions, Genetic ; genetics

The primers and probes for the Real-time RT-PCR were designed based on the multiple sequence (swine and humans HEV strains) alignments of the ORF3 region of genotype 4 HEV. A rapid, sensitive and stable TaqMan Real-time RT-PCR assay was established, and its specificity and sensitivity were assessed, and comparison of the Real-time RT-PCR with conventional and nested RT-PCR was performed. The results found that the crossing points showed linearly proportional to the logarithm of the input copy number. The correlation coefficient (R2) and the slope value of the standard curves with plasmid DNA were 0.994 and -3.312, respectively. The efficiency (E) of the PCR was 100%. Coefficients of variation values of the different diluted plasmid DNA were low in the same or different repeated experimental group. In addition, the assay was able to correctly detect genotype 4 HEV RNA from swine fecal samples. The sensitivity of established assay was 100-fold higher than that of conventional RT-PCR and 10-fold higher than nested RT-PCR.
Animals ; DNA Primers ; genetics ; Disease Reservoirs ; virology ; Feces ; virology ; Fluorescence ; Genotype ; Hepatitis E ; virology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Swine ; virology

OBJECTIVETo determine the seasonal prevalence of genotype-IV hepatitis E virus (HEV) in swine herds in Eastern China and explore the phylogenetic relationship between swine HEV and human HEV in the situation that zoonotic features of HEV had been confirmed.

METHODSFrom September 2007 to June 2008, a total of 1200 swine bile specimens were collected from three slaughter houses located in Zhejiang, Anhui and Jiangsu, the Eastern China, and detected for HEV RNA by using nested RT-PCR. The positive PCR products were sequenced. Then the swine HEV were phylogenetically determined with human HEV isolated in Eastern China.

RESULTSThe positive rate for HEV RNA in swine herds was 4.5% totally. Significant differences of HEV detection were not observed among seasonal pattern (Sep - Oct: 6%, Dec - Jan: 4.33%, Mar - Apr: 4.33%, May - Jun: 3.33%) but in geographic distribution (Jiangsu: 6%, Anhui: 5%, Zhejiang: 2.5%). Regardless of isolation from different areas,swine and human genotype-IV HEV shared a high similarity. Phylogenetically, there were 80% - 100% and 96% - 100% identities within swine genotype-lV HEV at the nucleotide and amino acid levels respectively. Between swine HEV and human HEV, there were also similarities of 76% -99% and 95% - 100%. It was noted that some human and swine isolates were clustered with bootstrap values of > 90%.

CONCLUSIONGenotype-IV HEV is widely prevalent in swine herds in Eastern China and original common ancestor of evolution and transmission was implied. The sustaining prevalence within swine herds should have a probable influence on the epidemic situation of hepatitis E in human beings.

Animals ; China ; epidemiology ; Genotype ; Geography ; Hepatitis E ; epidemiology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Phylogeny ; Prevalence ; Seasons ; Sequence Homology, Nucleic Acid ; Swine ; Swine Diseases ; epidemiology ; genetics ; virology

OBJECTIVETo investigate hepatitis E virus (HEV) infection among pigs in Henan province.

METHODSA total of 623 swine sera, collected from 5 districts, were divided into two groups, under 3-month of age and over 3-month of age. They were tested for HEV antigen and antibody by using ELISAs, respectively. The sera positive for HEV antigen were tested for HEV RNA with RT-PCR. The positive products of RT-PCR were cloned and sequenced.

RESULTSThe positive rates of anti-HEV antibody of the groups under 3-month and over 3-month of age were 90.27% and 92.55%, respectively, without statistical difference, while those of HEV antigen were 15.93% and 5.69%, respectively, with significant difference. The positive rates of anti-HEV antibody and HEV antigen were significantly different among different districts. HEV RNA was detectable in 5 of 47 HEV antigen positive samples. The sequence analysis showed that in 4 of 5 specimens the sequence belonged to genotype 4 while in the remaining one the sequence was genotype 1.

CONCLUSIONThe prevalence rate of HEV infection in pigs was high in Henan province and the rate differed in different districts.

Animals ; Antibodies, Viral ; analysis ; immunology ; Antigens, Viral ; analysis ; immunology ; China ; Genotype ; Hepatitis E ; epidemiology ; immunology ; veterinary ; virology ; Hepatitis E virus ; genetics ; immunology ; isolation & purification ; Phylogeny ; RNA, Viral ; analysis ; genetics ; Sequence Analysis, DNA ; Swine ; virology ; Swine Diseases ; epidemiology ; immunology ; virology

OBJECTIVETo determine the prevalence and genotype of hepatitis E virus (HEV) in swine liver before on the market and to analyze the phylogenetic relationship between the isolates from swine and human.

METHODS35 swine liver specimens were collected from two slaughtering houses in the countryside of Shandong province, China. Nested reverse transcription-polymerase chain reaction (nested RT-PCR) and subsequent sequencing were used to determine the nucleotide sequences. A phylogenetic tree was constructed with Neighbor-joining method based on the Kimura-2-parameter model.

RESULTS3. (8.57%) of the 35 swine liver specimens being tested were positive for HEV RNA. The three swine HEV strains isolated in the present study from liver samples shared the highest identity to genotype-IV HEV.

CONCLUSIONResults from the study confirmed that HEV was detectable among swine before on the market and the genotype was the same as that representing human and swine isolates in China. It also suggested that much more attention should be paid to the safety on the digestion of swine liver.

Abattoirs ; Animals ; China ; epidemiology ; Genotype ; Hepatitis E ; epidemiology ; virology ; Hepatitis E virus ; genetics ; isolation & purification ; Liver ; virology ; Phylogeny ; Prevalence ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, RNA ; Swine ; virology

BACKGROUND/AIMS: Most cases of hepatitis B virus (HBV) are transmitted vertically in endemic areas of HBV. The positivity of serum HBeAg/HBV DNA in pregnant women is associated with vaccine failure. Recently, a national program for HBV vaccines free of charge in neonates born to HBsAg-positive pregnant women is being performed. The aim of this study was to investigate the positivity of serological markers of HBV in pregnant women in Jeju, which is an island separated from the Korean peninsula and a promising cohort to evaluate the effect of a prevention program of HBV infection. In addition, we investigated the geographic differences in the prevalence of HBV infection because it has been reported that the prevalence of HBV has been high in this area previously. METHODS: Between January 2001 and December 2002, all women who gave delivery were studied retrospectively. Women between the ages of thirty and forty, who received health screening at the Asan Medical Center health promotion center in Seoul, were analyzed as controls. RESULTS: During the study period, 1,030 pregnant women (30.8 +/- 4.3 years) and 7,270 controls (33.1 +/- 5.0 years) were enrolled. The positivity of HBsAg was high in Jeju compared with that of Seoul (6.4% vs. 4.9%) (P=0.036). The positivity of HBeAg/HBV DNA was 31.8% (21/66) in HBsAg-positive pregnant women. The positivity of anti-HBs was low in Jeju compared with that of Seoul (54.5% vs. 68.8%) (P<0.001). CONCLUSIONS: The positivity of HBsAg was found to be high in pregnant women in Jeju. Intensive supervision for HBV infection in pregnant women should be given in this area.
Adult ; DNA, Viral/blood ; English Abstract ; Female ; Hepatitis B/diagnosis/*epidemiology ; Hepatitis B Surface Antigens/blood ; Hepatitis B e Antigens/blood ; Hepatitis B virus/genetics/isolation & purification ; Humans ; Korea/epidemiology ; Pregnancy ; Pregnancy Complications, Infectious/diagnosis/*epidemiology ; Seroepidemiologic Studies ; Serologic Tests