Case report: Role of hepatitis E virus in the etiology of community-acquired non-A,non-B hepatitis in greece

The aim of this study was to determine the frequency of hepatitis E virus (HEV) infection in a population of Greek adults with community-acquired (sporadic) non-A, non-B hepatitis found to be seronegative for antibodies to hepatitis C virus (anti-HCV). All patients admitted to the Liver Unit of Western Attica General Hospital and diagnosed as having acute community-acquired non-A, non-B hepatitis between February, 1986, and May, 1990, were enrolled in follow up studies (n = 66). Nineteen patients with HCV infection and 11 patients with acute non-A, non-B, non-C hepatitis that progressed to chronicity were excluded. Convalescent sera were tested for antibody to HEV (anti-HEV) by a fluorescent antibody blocking assay in 33 of 36 eligible patients. One of the 33 (3%) patients was found to be positive for anti-HEV. Anti-HEV testing of all 20 available serum specimens from this patient showed evidence of anti-HEV seroconversion at the fourth week after the onset of hepatitis. The patient had not travelled abroad or within Greece or had not had apparent contact with people from foreign countries for the previous 3 months. These data show that HEV infection is not a major cause of community-acquired non-A, non-B hepatitis in Greece. However, the reported case of HEV hepatitis suggests that HEV may retain a low endemicity in Greece. More extensive seroprevalence studies are needed for an accurate estimation of the extent of HEV infection in the southeastern European countries. © 1994 Wiiey-Liss, Inc.     

Detection of IgA class antibody to hepatitis E virus in serum samples from patients with hepatitis E virus infection

A newly developed assay for IgA class antibody to hepatitis E virus (IgA anti-HEV) was used to study 145 serum samples collected during an outbreak of an enterically transmitted hepatitis that occurred in 3 villages in the lower Shebeli region of Southern Somalia between January, 1988 and November, 1989. A total of 52.4% of the afflicted patients were found positive for IgA anti-HEV, and 73.1% of these were also positive for IgM. Both antibodies disappeared during the convalescence period. Similar results were also seen in serum obtained from sporadic cases of acute waterborne hepatitis in Pakistan. © 1993 Wiley-Liss, Inc.     

Serological studies of an enterically transmitted non-A,non-B hepatitis in Somalia

An outbreak of an acute sporadic viral hepatitis occurring at three villages in the lower Shebli region of southern Somalia was studied. Sera were examined for antibodies to hepatitis E virus (HEV) antigen by a recently developed enzymelinked immunosorbent assay. The prevalence of anti-HEV ranged from 77.8% to 94.0% among the three villages. Anti-HEV prevalence did not differ significantly with age or sex. HEV infections were widely distributed among all age groups, without a preponderance in any specific age category. Overall, 97.6% of afflicted patients who were examined 6–10 days after onset of jaundice developed either IgM or IgG anti-HEV. © 1993 Wiley-Liss, Inc.     

Prevalence of antibodies to hepatitis E virus in residents of a district in Havana, Cuba

A seroepidemiological study of hepatitis E virus (HEV) infection was conducted in a district of Havana, where hepatitis A virus (HAV) is considered endemic. The levels of anti-HEV antibodies were evaluated by enzyme-linked immunosorbent assay (ELISA) based on the recombinant protein GST-ORF2.1. Anti-HEV antibodies were detected in 11 of 209 (5.3%) of serum samples, compared to 71.3% for anti-HAV antibodies. No risk factors reported previously for HEV infection showed a significant association with the presence of anti-HEV antibodies, whereas anti-HAV antibodies were strongly associated with increasing age. HEV may be considered endemic in this area and is likely to have a significant clinical impact.     

血清学标志阴性的非甲～戊型肝炎的病原学研究

目的对血清学标志阴性的非甲～戊型肝炎进行病原学研究。方法用ＨＢＶＰＣＲ、ＨＣＶＲＴ－ＰＣＲ和ＨＥＶＲＴ－ＰＣＲ分别检测血清学标志阴性的非甲～戊型肝炎患者血清，并对其部分阳性产物进行克隆测序。结果８７例非甲～戊型肝炎血清ＨＢＶＤＮＡ均为阴性，９例（１０．３％）为ＨＣＶＲＮＡ阳性，部分经测序证实为ＨＣＶ１ｂ亚型；余７８例为ＨＢＶＤＮＡ和ＨＣＶＲＮＡ均阴性。该７８例中，１４例因无血清未作ＨＥＶＲＮＡ检测，余６４例中４９例（７６．６％）为ＨＥＶＲＮＡ阴性，１５例（２３．４％）为ＨＥＶＲＮＡ阳性。经序列分析显示，其中９例为典型的中国ＨＥＶ株基因序列，６例变异较大，与典型的中国株基因序列的同源性仅为８０％左右。４９例ＨＢＶＤＮＡ、ＨＣＶＲＮＡ和ＨＥＶＲＮＡ均阴性的血清中１６例（３２．６％）ＨＧＶＲＮＡ阳性。由此可见，该８７例中至少有９例为ＨＣＶ感染，１５例为ＨＥＶ感染，１６例为ＨＧＶ感染。结论对血清学标志阴性的非甲～戊型肝炎的病人应该用ＰＣＲ法进行病原学分型，以明确其诊断     

Seroprevalence of hepatitis E virus antibodies in different human populations of Cameroon

Hepatitis E virus (HEV) is a major cause of acute hepatitis worldwide. HEV infection is self-limiting, but fulminant hepatitis may occur with higher mortality rates. The aim of this study was to determine the seroprevalence of the HEV in three different populations in Cameroon and to assess alimentary risk factors of infection. A total of 903 individuals including 450 elderly people, 183 pregnant women, and 270 HIV-infected patients were enrolled during 2009-2015. All sera obtained were tested for the presence of anti-HEV antibodies with commercial enzyme-linked immunosorbent assay kits. The association between initial anti-HEV status and potential risk factors was assessed. Out of the 903 samples, 22.0% (199/903) were positives for anti-HEV immunoglobulin M (IgM), 5.8% (52/903) for anti-HEV IgG, and 3.5% (32/903) for both. The seropositivity for IgM was 7.0%, 13.1%, and 34.7%; meanwhile, the seropositivity for immunoglobulin G was 8.5%, 3.3%, and 5.1%, in HIV-infected patients, pregnant women, and the elderly population, respectively. Both antibodies were detected simultaneously in 2.2%, 1.6%, and 5.1% in HIV-infected patients, pregnant women, and the elderly population, respectively. No risk factors were significantly associated with HEV infection in these populations. This study showed a high prevalence of anti-HEV antibodies in three different populations in Cameroon.     

Full‐length sequence of genotype 3 hepatitis E virus derived from a pig in Thailand

Hepatitis E virus (HEV) infection in pigs was investigated in two principal swine farming areas in Thailand. Anti‐HEV antibodies and HEV RNA in sera were examined in 258 pigs reared on five commercial farms from age 1 to 6.5 months and sows. Overall, 167 of 258 (64.7%) pigs were positive for anti‐HEV IgG, while 20 of 258 (7.75%) had detectable HEV RNA. Sequence analysis of 20 HEV isolates obtained from viremic pigs revealed that they were 92.3–100% identical to each other and had 82.2–88.2% nucleotide similarity to other reported genotype 3 isolates in 415 nucleotide sequences within ORF2 region. Further characterization by sequencing the complete genome of the Thai swine HEV isolate (named Thai‐swHEV07) and phylogenetic analysis showed that Thai‐swHEV07 segregated into a cluster consisting of swine isolates from Japan, Mongolia, and Kyrgyzstan within the HEV genotype 3. The Thai‐swHEV07 had a genomic length of 7,229 nt excluding the polyadenylated region at 3′ terminus of the genome. Comparison of Thai‐swHEV07 and 27 reported strains of genotype 3 revealed 80.4–85.9% nucleotide identity, with the highest identity of 85.9% to the novel swHEV strain from Mongolia. These findings suggest that genotype 3 HEV isolates are markedly heterogeneous. J. Med. Virol. 81:657–664, 2009 © 2009 Wiley‐Liss, Inc.     

Antibodies to hepatitis E virus among chinese patients with acute hepatitis in Taiwan

The prevalence of antibodies to hepatitis E virus (anti-HEV) was investigated in patients with acute hepatitis, and correlated with the clinical features. Sera from 110 patients with acute hepatitis and 60 healthy controls were tested for anti-HEV, antibody to hepatitis C virus (anti-HCV), and hepatitis B surface antigen (HBsAg). There were significant differences in the prevalence of anti-HEV, anti-HCV, and HBsAg between patients and controls (21.8% vs. 0%, 16.3% vs. 1.6% and 58.1% vs. 18.0%, respectively). Anti-HEV was detected in 6 (25.0%) of 24 patients with anti-HCV, 6 (9.3%) of 64 patients with HBsAg, and another 6 (22.2%) of 27 patients with acute hepatitis non-A, non-B, non-C. Anti-HEV was found in 15 men and three women, whose ages ranged from 34 to 75 (median, 57) years old. The median age of patients with anti-HEV was older than that in patients without this antibody (57 vs. 38 years; P = 0.001). The prevalence of anti-HEV in patients with anti-HCV alone (35.2%) was higher than that (11.1%) in patients with HBsAg alone (P = 0.03). Compared to patients without anti-HEV, HEV-infected patients had a higher frequency of travel to a foreign country (P = 0.0001), had a lower HBsAg rate (P = 0.019), and had higher serum alkaline phosphatase levels (P = 0.04) and gamma-glutamyl transpeptidase levels (P = 0.01). In conclusion, HEV infection occurs in 22.2% of patients with acute hepatitis non-A, non-B, non-C. HEV superinfection may occur in patients with chronic hepatitis B or C virus infection. © 1994 Wiley-Liss, Inc.     

Interference between non-A, non-B and hepatitis B virus infection in chimpanzees

Pre-existing chronic or concurrent acute non-A, non-B (NANB) virus infection has been found to interfere with hepatitis B virus (HBV) replication and to delay and moderate markedly the appearance of disease related to HBV infection in chimpanzees. The mechanism for this phenomenon remains unclear. These findings are of practical significance for vaccine safety testing and evaluation of methods for virus inactivation in chimpanzees. The possible occurrence of dual NANB and HBV infection requires that prolonged follow-up be carried out. Attempts to carry out inactivation studies on materials containing both types of virus may also provide misleading observations.     

TT virus infection in healthy children,children after blood transfusion,and children with non-A to E hepatitis or other liver diseases in Taiwan

Serum samples from healthy and diseased children were studied for the presence of TTV DNA by nested PCR using primer sets generated from N-22 region and from the untranslated region (UTR) of the viral genome. N-22 positive TTV DNA was detectable in 33 (27%) of 122 healthy children, 47 (73.4%) of 64 polytransfused thalassemic children, 37 (46.3%) of 80 children who received transfusion during cardiac surgery, 8 (42.1%) of 19 non-A to E hepatitis, 10 (33.3%) of 30 HBV carrier children, and 5 (15.6%) of 32 infants with biliary atresia. A much higher prevalence of TTV DNA with rates varying from 78-100% in the above study groups was observed using the UTR primers. For children with N-22 positive TTV DNA, biochemical assessment of isolated TTV viremia in thalassemic children or children transfused during surgery showed no convincing association between raised ALT levels and TTV viremia. Coinfection with TTV in chronic HCV-infected or HBV-infected children did not result in higher peak ALT levels during follow-up, suggesting that TTV has no synergistic pathogenic effect. The phylogenetic analysis of the N-22 positive TTV DNA isolates revealed that most isolates from healthy children, children transfused during surgery, and non-A to E fulminant hepatitis children were type 1 TTV. These results indicate that TTV infection in children was significantly associated with transfusion. TTV infection is highly prevalent in early childhood in Taiwan but plays a minimal role in the induction of hepatitis in children.     

Valuable antibody detection method for classifying hepatitis E virus genotypes

Nucleotide‐based methods are conventionally used to classify the hepatitis E virus (HEV) genotypes. A serological enzyme immunoassay (EIA) using open reading frame 3 (ORF3) C‐terminal peptides was developed to conveniently and accurately classify and evaluate the genotypes of HEV. The sera of mice immunized with HEV genotype 1, 3, and 4 reacted highly specifically to the peptides of the corresponding genotypes. Most (84.2%) clinical sera infected with HEV genotype 4 were positive for anti‐HEV antibodies when tested with the ORF3 peptides of genotype 4, but were negative for genotypes 1 and 3. Monkey and clinical serial sera infected with HEV reacted strongly to the homologous genotype ORF3 peptides. The indirect EIAs were more sensitive, with stronger reactivity, than commercial anti‐HEV immunoglobulin G assays when serial sera from monkeys infected with HEV genotype 1 or 4 were tested. All our results indicate that the serological typing EIA assays described in this study are more effective and convenient for the classification of HEV genotypes than molecular approaches, and can be used to screen large numbers of serum samples and differentiate genotypes for the diagnosis of HEV infections.     

Assay of HBV DNA in the plasma of HBV-carrier chimpanzees superinfected with non-A, non-B hepatitis

A dot hybridisation technique was used to monitor the levels of hepatitis B virus (HBV) DNA in the plasma of two HBV-carrier chimpanzees which had been inocula ed with documented infectious non-A, non-B hepatitis agents. A marked decrease in the quantity of HBV DNA in the plasma during the acute phase of the non-A, non-B hepatitis was observed in both carriers. The possible role of interferon or a similar antiviral agent in modulation of the HBV-carrier state is discussed. Hybridisation may become, in due course, the method of choice for examining blood samples for infectious hepatitis B virus.     

Investigation of hepatitis E virus infection in swine from Hunan province, China

Nine hundred and four serum samples were collected from pigs from 16 pig farms in China's Hunan province and tested for anti-hepatitis E virus (HEV) antibody and the HEV capsid antigen using EIAs. Of the 904 samples, 617 (68.3%) and 57 (6.3%) were positive for anti-HEV antibody and antigen, respectively. The prevalence of HEV antibodies and detection of antigen varied significantly among pigs from different farms (P < 0.01). The positivity rate for anti-HEV antibody and occurrence of high titer antibody were significantly higher in pigs above, than in those below, 3 months of age, whilst the detection of antigen did not differ significantly between the two groups. Based on these data, 481 serum samples that were positive for HEV antigen or with an S/CO < or =10 for anti-HEV were tested for HEV RNA using real-time RT-PCR; 28 of 481 (5.8%) were positive. The positivity rate of HEV RNA was much higher for the HEV antigen positive sera (40.1%) than the anti-HEV antibody positive (1.4%) or negative (1.1%), antigen negative samples. Sixteen of the 28 samples were positive for HEV RNA using nested RT-PCR and their products were cloned and sequenced. These 16 isolates belonged to HEV genotype 4, including 12 that did not belong to any subtype of HEV genotype 4 reported previously. Thus, the infection rate of HEV in pigs is high and the HEV antigen has a close relationship with HEV RNA. The HEV genotype infecting the pigs was genotype 4 and a novel subtype may exist in Hunan province.     

Immune response to GOR,a marker for non-A,non-B hepatitis and its correlation with hepatitis C virus infection

Recently, identification and molecular cloning of a host cellular gene designated GOR from chimpanzees experimentally infected with non-A, non-B hepatitis (NANBH) agent was reported. It was further demonstrated that there is a close association between the immune response to an antigenic peptide of GOR (GOR2) and NANBH. In order to define the specificity of the immune response, in the present study we have identified an additional epitope in the GOR gene sequence, upstream from GOR2, and studied its correlation with the immune response to hepatitis C virus (HCV) in NANBH patients. An enzymelinked immunoassay (EIA) was developed which utilizes synthetic peptides designated spGOR346 and spGOR2 as the serological target for the detection of anti-GOR antibodies in patient serum samples from various hepatic and non-hepatic disease categories. GOR peptides identified 80-90% of the NANBH samples that were positive for HCV C100-3 and about 70% of the NANBH samples that were positive by Abbott prototype second-generation HCV antibody assay. Among a normal donor population(s), only 2-3% of the samples were positive for antibodies to GOR sequences, whereas from the patient categories unrelated to viral hepatitis as well as various nonhepatic diseases, the immune response to both GOR peptides was closely associated with the presence of antibodies to HCV. The data indicate that antibodies to GOR is a marker associated with NANBH.