HEV RNA Research Articles

The prevalence of HEV among non-A-C hepatitis in Qatar and efficiency of serological markers for the diagnosis of hepatitis E

BackgroundThe rapid growth of Qatar in the last two decades has attracted a large influx of immigrant workers who mostly come from HEV-hyperendemic countries. Thus, we aim to investigate the prevalence of HEV among acute non-A-C hepatitis patients in Qatar; and to evaluate the performance of four dominant commercial serological assays for HEV diagnosis.Methods259 patients with non-A-C hepatitis were tested using the Wantai HEV-IgM, HEV-IgG, HEV-Ag ELISA kits, and the MP Biomedical HEV-Total Ab ELISA kit. ALT levels were tested and HEV RNA (viral loads) was performed using Taqman AmpliCube HEV RT-PCR kit (Mikrogen, Neuried, Germany). The performance of each kit was assessed according to the RT-PCR results.ResultsHEV-RNA was detected in 23.1% of the samples. Most of these HEV-RNA-positive cases belonged to non-Qatari residents from the Indian subcontinent; India, Pakistan, etc. HEV-Ag, HEV-IgM, HEV-IgG, HEV-Total Ab were detected in 5.56%, 8.65%, 32.1%, and 34.2% of all tested samples, respectively. Elevated ALT levels were highly correlated with the HEV-Ag, HEV-IgM, HEV-RNA but not with the HEV-IgG and HEV-Total Ab. Although HEV-Ag was very specific (100%), yet its sensitivity was poor (36.7%). HEV-IgM demonstrated the best second marker for diagnosis of acute HEV after RT-PCR as jugged by the overall performance parameters: specificity (96.2%), sensitivity (71.4%), PPV (83.3%), NPP (92.7%), agreement with RT-PCR (91.0%), and Kappa-value (0.71).ConclusionOur study demonstrated a high prevalence of HEV virus in Qatar, mostly among immigrants from the Indian subcontinent. The HEV-IgM represents the best marker for detecting the acute HEV infection, where RT-PCR cannot be performed.

Hepatitis E Virus RNA Presence in Wild Boar Carcasses at Slaughterhouses in Italy.

Simple SummaryHepatitis E virus (HEV) is a worldwide diffused pathogen responsible for acute hepatitis of humans. Transmission of the pathogen is mostly related to the consumption of contaminated food and water. Although initially the disease was contained in developing countries, in recent years autochthonous infections have been reported in several industrialised countries. A different epidemiological pattern of transmission has been highlighted; while in Africa and Asia transmission is mainly due to waterborne outbreaks caused by low sanitation standards, in Europe and other industrialised countries, the disease has mainly spread due to consumption of raw or undercooked meat and seafood. Although HEV has been identified in several domestic and wild animal species, pigs and wild boar, appear to play a distinct role mainly acting as a reservoir of the pathogen. In this study, we monitored the presence of HEV in carcasses and livers of wild boar sampled in Tuscany at the slaughterhouse following hunting activities. Our data indicate the presence of the pathogen in the liver and the carcasses, suggesting cross-contamination. This evidence highlights the importance of maintaining safety control measures to avoid the spreading of HEV infection.Hepatitis E virus (HEV) is a waterborne and foodborne pathogen largely spread around the world. HEV is responsible for acute hepatitis in humans and it is also diffused in domestic and wild animals. In particular, domestic pigs represent the main reservoir of the infection and particular attention should be paid to the consumption of raw and undercooked meat as a possible zoonotic vehicle of the pathogen. Several studies have reported the presence of HEV in wild boar circulating in European countries with similar prevalence rates. In this study, we evaluated the occurrence of HEV in wild boar hunted in specific areas of Tuscany. Sampling was performed by collecting liver samples and also by swabbing the carcasses at the slaughterhouses following hunting activities. Our data indicated that 8/67 (12%) of liver samples and 4/67 (6%) of swabs were positive for HEV RNA. The presence of HEV genome on swabs indicates the possible cross-contamination of carcass surfaces during slaughtering procedures. Altogether, our data indicated that it is essential to promote health education programmes for hunters and consumers to limit the diffusion of the pathogen to humans.

Evaluation of hepatitis E antigen kinetics and its diagnostic utility for prediction of the outcomes of hepatitis E virus genotype 1 infection

ABSTRACT HEV-Ag ELISA assay is a reliable diagnostic test in resource-limited areas. HEV genotype 1 (HEV-1) infections are either self-limited or progress to fulminant hepatic failure (FHF) and death if anti-HEV therapy is delayed. Limited data is available about the diagnostic utility of HEV Ag on HEV-1 infections. Herein we aimed to study the kinetics of HEV Ag during HEV-1 infections at different stages, i.e., acute HEV infection, recovery, and progression to FHF. Also, we evaluated the diagnostic utility of this marker to predict the outcomes of HEV-1 infections. Plasma of acute hepatitis E (AHE) patients were assessed for HEV RNA by RT-qPCR, HEV Ag, and anti-HEV IgM by ELISA. The kinetics of HEV Ag was monitored at different time points; acute phase of infection, recovery, FHF stage, and post-recovery. Our results showed that the level of HEV Ag was elevated in AHE patients with a significantly higher level in FHF patients than recovered patients. We identified a plasma HEV Ag threshold that can differentiate between self-limiting infection and FHF progression with 100% sensitivity and 88.89% specificity. HEV Ag and HEV RNA have similar kinetics during the acute phase and self-limiting infection. In the FHF stage, HEV Ag and anti-HEV IgM have similar patterns of kinetics which could be the cause of liver damage. In conclusion, the HEV Ag assay can be used as a biomarker for predicting the consequences of HEV-1 infections which could be diagnostically useful for taking the appropriate measures to reduce the complications, especially for high-risk groups.

Hepatitis E in solid organ transplant recipients: A systematic review and meta-analysis.

BACKGROUNDHepatitis E virus (HEV) infection is underdiagnosed due to the use of serological assays with low sensitivity. Although most patients with HEV recover completely, HEV infection among patients with pre-existing chronic liver disease and organ-transplant recipients on immunosuppressive therapy can result in decompensated liver disease and death.AIMTo demonstrate the prevalence of HEV infection in solid organ transplant (SOT) recipients. METHODSWe searched Ovid MEDLINE, EMBASE, and the Cochrane Library for eligible articles through October 2020. The inclusion criteria consisted of adult patients with history of SOT. HEV infection is confirmed by either HEV-immunoglobulin G, HEV-immunoglobulin M, or HEV RNA assay.RESULTSOf 563 citations, a total of 22 studies (n = 4557) were included in this meta-analysis. The pooled estimated prevalence of HEV infection in SOT patients was 20.2% [95% confidence interval (CI): 14.9-26.8]. The pooled estimated prevalence of HEV infection for each organ transplant was as follows: liver (27.2%; 95%CI: 20.0-35.8), kidney (12.8%; 95%CI: 9.3-17.3), heart (12.8%; 95%CI: 9.3-17.3), and lung (5.6%; 95%CI: 1.6-17.9). Comparison across organ transplants demonstrated statistical significance (Q = 16.721, P = 0.002). The subgroup analyses showed that the prevalence of HEV infection among SOT recipients was significantly higher in middle-income countries compared to high-income countries. The pooled estimated prevalence of de novo HEV infection was 5.1% (95%CI: 2.6-9.6) and the pooled estimated prevalence of acute HEV infection was 4.3% (95%CI: 1.9-9.4).CONCLUSIONHEV infection is common in SOT recipients, particularly in middle-income countries. The prevalence of HEV infection in lung transplant recipients is considerably less common than other organ transplants. More studies examining the clinical impacts of HEV infection in SOT recipients, such as graft failure, rejection, and mortality are warranted.

Seroprevalence of hepatitis E virus in risk populations and blood donors in a referral hospital in the south of Brazil

The prevalence of anti-hepatitis E virus (HEV) antibodies has a high heterogeneity worldwide. South American data are still scarce. The aim of this study was to evaluate the prevalence of HEV in populations at risk in comparison to blood donors (BD). A cross-sectional study was carried out in adults of different risk populations including crack users (CK), residents in a low income area (LIA), cirrhotic (CIR) and liver transplant patients (LT) compared with BD. The WANTAI HEV ELISA test was used and real-time PCR (in-house for screening and ALTONA as confirmatory test) for HEV RNA screening. A total of 400 participants were included. Anti-HEV IgG was positive in 19.5% of the total sample, reaching the highest rate in the CIR group, 22.5%, followed by CK, LT, and LIA (20%, 18.7%, and 17.5%, respectively). The prevalence found in BD individuals was of 18.7% (p = NS). Anti-HEV IgM was positive in only 1.5% of the sample (6/400). No blood or stools samples were positive for HEV RNA. The seroprevalence reported is among the highest rates ever found in Brazil. Considering the intense diagnostic investigation, data show that HEV circulation is more common that might be expected in our country.

Anti-HEV IgG Avidity Testing: Utility for Diagnosing Acute and Resolved Genotype 3 Infections.

European Association of the Study of the Liver (EASL) guidelines specify HEV RNA, as well as anti-HEV IgG and IgM as positive markers for acute HEV infection. HEV RNA assay sensitivity limitations may lead to false negative test results in patients with low levels of viremia. Moreover, anti-HEV IgM positivity is not a reliable indicator for distinguishing between acute and resolved infections given the ability of this antibody to persist several months after a resolved infection. Our study aims were to assess HEV IgG avidity for diagnosing acute and resolved infections, regardless of the anti-HEV IgM serostatus, and examine assay reliability when evaluating different genotype 3 (GT3) HEV subtypes. Patient serum samples (n = 104) were tested for HEV IgG avidity by utilizing the DIA.PRO kit on a DSX automated instrument. Among patients identified with acute HEV infections, 32 were infected with GT3: GT3c (n = 5), GT3e (n = 8), 3f (n = 17) and GT3-unsubtyped (n = 2). Avidity sensitivity was 91.2% and specificity was 100%. For patients with long-lasting anti-HEV IgM persistence, an Avidity Index >70% was observed. Thus, the DIA.PRO avidity assay may be utilized to distinguish between recently acquired and resolved HEV GT3 infections. However, for equivocal results (Avidity Index > 40–70%), HEV RNA molecular testing will be required to confirm a recent infection.

Laboratory-based Surveillance and Clinical Profile of Sporadic HEV Infection in Shanghai, China.

The study aimed to describe the epidemiological, virological and clinical features of sporadic HEV infection in eastern China. A total of 6112 patient sera were tested for anti-HEV IgG or anti-HEV IgM during one consecutive year (between August 2018 and July 2019). HEV RNA presence was evaluated by RT-PCR and HEV sequences were phylogenetically analyzed. Clinical features of confirmed HEV-infected patients were delineated. The sero-positivity rate of anti-HEV IgG maintained stable around 40%, while an obvious winter spike of anti-HEV IgM prevalence was observed. A total of 111 patients were confirmed of HEV viremia by molecular diagnosis. Subtype 4d was predominant. Phylogenetic analyses suggest that certain strains circulate across species and around the country. Subjects with confirmed current HEV infection had a high median age (58years) and males were predominant (62.2%). Most patients presented with jaundice (75.7%) and anorexia (68.0%). Significantly elevated levels of liver enzymes and bilirubin were observed. Remarkably, the baseline bilirubin level was positively correlated with illness severity. Pre-existing HBV carriage may deteriorate illness. The clinical burden caused by locally acquired HEV infection is increasing. Surveillance should be enforced especially during the transition period from winter to spring. Patients with higher level of bilirubin at disease onset had slower recovery from HEV infection.

Role of NLRP3 inflammasome in the immunopathogenesis of HEV infected AVH and ALF patients

Background: Hepatitis E virus is one of the leading causes of acute viral hepatitis with worldwide distribution amounting to 3.3 million symptomatic cases and ∼60,000 deaths. 30–45% of acute liver failure (ALF) patients in India are positive for hepatitis E virus marker. High mortality (∼30%) in HEV infected pregnant patients has been reported. The pathogenesis of the disease is less understood, thus the present study focuses on the role of inflammasomes and pro-inflammatory cytokines in the immunopathogenesis of HEV. Methods and materials: 102 out of 1680 suspected cases were positive for HEV IgM and/or HEV RNA. 30 AVH and 15 ALF HEV positive sera were subjected for IL-1β, IL-18 and IL-1BPa cytokine and viral load estimation. PBMCs were isolated and processed for mRNA expressions of NLRs (NLRP1, LRP3, NLRC4); RLRs (RIG-1, MDA5, LGP2); cytokines (IL-1β, IL-18) and other necessary inflammasome pathway genes (CASP1, CASP3, PYCARD, NF-κβ). Mechanism of cell death in the PBMCs of HEV infected patients was studied by Annexin/PI apoptosis kit. Results: Lower limit of detection and quantification by HEV TaqMan Real-Time PCR was estimated as 9 and 30 viral copies/μl of sample respectively. High viral load was observed in ALF than AVH cases (1.55 × 105 ± 1.30 × 105 vs 8.18 × 103 viral copies/μl). mRNA expression of NLRP3 gene in AVH was positively correlated with IL-18 (r = 0.74, p ∼ 0.0001***) and IL-1β (r = 0.68, p ∼ 0.0001***). Significant levels of serum IL-1β and IL-18 were observed in AVH as compared to ALF patients (7.53 ± 3.92 pg/ml vs 3.39 ± 1.37 pg/ml; 1905 ± 751.9 pg/ml vs 1128 ± 705.2 pg/ml) respectively. Interleukin-18 binding protein a (IL-18BPa) levels were significantly high in AVH (24,960 ± 2351 pg/ml) in comparison to ALF (21,000 ± 2796 pg/ml) with the levels of 4960 ± 1639 pg/ml in healthy controls. Annexin/PI assay showed 36.82 ± 5.98% apoptotic cell death in AVH in comparison to healthy control. However no significant difference was observed between AVH and ALF patients. Conclusion: Significant upregulation of NLRP3 inflammasome leading to high production of IL-18 and IL-1β cytokines in sera of AVH patients indicated the role of TH1 response acting through NLRP3 pathway which might be helpful in recovery of AVH patients.

Persistent Hepatitis E virus infection across England and Wales 2009-2017: Demography, virology and outcomes.

The first clinical case of persistent HEV infection in England was reported in 2009. We describe the demography, virology and outcomes of patients identified with persistent HEV infection in England and Wales between 2009 and 2017. A series of 94 patients with persistent HEV infection, defined by HEV viraemia of more than 12weeks, was identified through routine reference laboratory testing. Virology, serology and clinical data were recorded through an approved PHE Enhanced Surveillance System. Sixty-six cases (70.2%) were transplant recipients, 16 (17.0%) had an underlying haematological malignancy without stem cell transplantation, six (6.4%) had advanced HIV infection, five (5.3%) were otherwise immunosuppressed, and one patient (1.1%) had no identified immunosuppression. Retrospective analysis of 46 patients demonstrated a median 38weeks of viraemia before diagnostic HEV testing. At initial diagnosis, 16 patients (17.0%) had no detectable anti-HEV serological response. Of 65 patients treated with ribavirin monotherapy, 11 (16.9%) suffered virological relapse despite undetectable RNA in plasma or stool at treatment cessation. Persistent HEV infection remains a rare diagnosis, but we demonstrate that a broad range of immunocompromised patients are susceptible. Both lack of awareness and the pauci-symptomatic nature of persistent HEV infection likely contribute to significant delays in diagnosis. Diagnosis should rely on molecular testing since anti-HEV serology is insufficient to exclude persistent HEV infection. Finally, despite treatment with ribavirin, relapses occur even after cessation of detectable faecal shedding of HEV RNA, further emphasising the requirement to demonstrate sustained virological responses to treatment.

An outbreak of hepatitis E in Yavatmal, India, 2019.

Hepatitis E, a public health concern in developing countries, frequently presents in epidemic, as well as in sporadic forms. This study investigated an outbreak of viral hepatitis at Yavatmal, Maharashtra, India in March 2019. Blood samples from 10 patients were received at Indian Council of Medical Research-National Institute of Virology, Pune to test for the presence of enterically transmitted hepatitis viruses. Subsequently, 49 suspected cases were screened for anti-hepatitis E virus (HEV)/hepatitis A virus (HAV) immunoglobulin M and immunoglobulin G (IgG) antibodies, alanine amino-transferase levels and HEV RNA. Water samples were screened for HEV and HAV RNA followed by phylogenetic analysis. Overall 32 of 49 (65.3%) suspected cases had recent acute HEV infection, while dual infection with HAV was noted in one case (2.04%). Forty-eight of 49 suspected cases were positive for anti-HAV IgG antibodies indicative of previously acquired immunity against HAV. Water samples had evidence of HEV contamination as detected by reverse transcription-polymerase chain reaction. Sequencing of HEV RNA from both patients (n = 2) and water samples (n = 5) indicated HEV genotype 1 to be the etiological agent of this outbreak. Serological and molecular evidence confirmed HEV as the etiology. Mixing of contaminated drain water with the domestic water supply may have triggered this outbreak. Subsequent changing of the defaulted water pipelines and its segregation from drain pipelines by the health authorities resulted in progressive decline of this outbreak. Despite the limitations, periodic surveillance of HEV exposure pattern and reporting of small outbreaks would supplement to the global disease burden data of hepatitis E.

Identification of Hepatitis E Virus in the Feces of Red Foxes (Vulpes vulpes).

Orthohepeviruses (HEV) can infect a wide range of animals, showing a relatively strict host specificity; however, its zoonotic potential, natural transmission in the wildlife are less known. Several new HEV-like viruses have been identified in various animal species, including carnivores; however, the phylogenetic relationship among these viruses is poorly resolved, since some of them were known as rodent-related so far. The red fox, the most widespread carnivore worldwide, is a known reservoir of several viruses that transmit from wildlife to humans or domestic animals; they might have a defined role in the circulation of rodent-borne HEV. In this study, we performed a HEV survey by heminested RT-PCR (Reverse Transcription PCR) on red fox fecal samples to investigate the presence of HEV in red foxes living in natural conditions, and to explore the origin of the virus via phylogenetic analysis. Out of the 26 investigated samples, HEV RNA was identified in one sample. Following Sanger sequencing, the novel sequence displayed 91% identity on the nucleotide level with recently published European common vole-HEV derived from Microtus arvalis. In contrast, it shared 85% nucleotide similarity with HEV strains described previously in red foxes. Our results strongly support "the dietary-origin" of unclassified HEV-like strains described from predators that usually prey on rodents.

Phylogenetic analysis and epidemiological history of Hepatitis E virus 3f and 3c in swine and wild boar, Italy

BackgroundHepatitis E virus (HEV) genotype 3 has a worldwide distribution. The food-borne transmission of HEV associated with the consumption of products derived from domestic pig, wild boar has been reported in various countries. In this study the genetic diversity, evolutionary rates of HEV 3f, 3c among swine and wild boar in Italy were estimated. MethodsSampling was performed on a wild boar population living in an area located in Abruzzo region. The HEV RNA amplification was performed by real-time RT-PCR. Nested RT-PCR and sequencing of the ORF2 region were carried out by the Super Script III First-Strand Synthesis System. Sequencing of purified PCR products was carried out by the Genome Lab Dye Terminator Cycle Sequencing (DTCS) Quick Start Kit. The maximum likelihood trees were generated by using Phyml. The mean evolutionary rates and the dated trees were co-estimated by BEAST. ResultsThe phylogenetic analysis showed that the HEV ORF2 isolates from Abruzzo region belonged to 3f subtype. The prevalent subtypes in Italy were those belonging to 3f and 3c. The estimated mean values of the HEV ORF2 capsid gene evolutionary rates were 1.915 × 10−2 substitutions/site/year (95% HPD: 1.64 × 10−3 – 3.97 × 10−2) and 2.81 × 10−2 substitutions/site/year (1.83 × 10−2 - 3.8 × 10−2) for 3f and 3c subtype datasets, respectively.The HEV 3f dated back to 1985 (1960–2000), whereas the 3c subtype entered in Italy during the year 2006 (2005–2006). The majority of the HEV 3f sequences collected from swine didn't appear intermixed, except in two cases. The HEV 3c population circulating in Italy remained segregated without significant transfer to swine. ConclusionOur study provide insight into the evolution, circulation of HEV 3f and 3c in Italy.Continued genomic surveillance of HEV in animal reservoir, as well as improving sanitary control measures are required.

Characterization of hepatitis E virus natural infection in farmed rabbits.

Rabbit hepatitis E virus (HEV3-ra) is widely distributed in rabbits worldwide and several recent reports found that HEV3-ra can infect humans. Therefore, people exposed to rabbits are at high risk of HEV infection. This study was conducted to investigate the characteristics and outcomes of HEV3-ra natural infection in rabbits. Seventy farmed rabbits (3-month-old) were surveyed in a farm in Beijing, China. Rabbits tested positive for HEV RNA were followed weekly for testing of HEV RNA, antigen, antibody and alanine aminotransferase (ALT) level. Liver and kidney tissue was collected for histopathology. Complete genome sequencing of the isolated HEV3-ra strain was performed (CHN-BJ-r4, GenBank: MT364355). The infectivity of CHN-BJ-r4 was tested in ten naïve rabbits by intravenous injection or gavage. Anti-HEV antibody and HEV RNA were tested positive in 7.14% (5/70) and 11.4% (8/70) of rabbits, respectively. Eight naturally infected rabbits were followed, and 37.5% (3/8) of the observed rabbits were found to have fecal shedding of HEV ranging from 3-22weeks with high viral load (105 -107 copies/g). Two out of eight rabbits showed temporary viremia. Naturally infected rabbits presented elevated ALT level, seroconversion, and liver histopathology. Complete genome of HEV3-ra isolated in this study shared 84.61%-94.36% nucleotide identity with known HEV3-ra complete genomes. The isolated HEV3-ra strain was infectious and could infect other rabbits through intravenous and fecal-oral route. Naturally infected rabbits showed up to 22-week fecal virus shedding with high viral load. These features increased the risk of rabbit-to-rabbit and rabbit-to-human transmission.

Hepatitis E 3ra Genotype Infection in People Living With HIV in Spain.

BackgroundThe objective of our study was to assess the prevalence and incidence of HEV in people living with HIV (PLWH) in a Spanish national cohort.MethodsRetrospective longitudinal study including PLWH recruited in the cohort of adult HIV-infected patients of the AIDS Research Network in follow-up at 28 Spanish hospitals with available serum samples in 2014 and 2015. All samples were tested for HEV IgG, IgM, and RNA. Samples with detectable HEV viral loads were genotyped. Prevalence and incidence of HEV infection were calculated.ResultsThe study sample comprised 845 PLWH. At baseline, 101 patients were positive for HEV IgG antibodies (11.9%), none had HEV IgM antibodies, and 2 presented detectable HEV RNA (0.23%). Forty-two seroconverted for IgG, supposing a cumulative incidence of 5.7%. One subject was positive for IgM (0.13%), and 2 showed detectable HEV RNA (0.27%). One case was infected by the emergent HEV genotype 3ra.ConclusionOur study identifies one case of HEV 3ra genotype infection, the main host of which is rabbit, showing a potential zoonotic role of this emerging genotype in Spain.

First report of HEV-3 subtypes e and f in the wild boar population in Abruzzo region, Italy

Abstract Background Hepatitis E virus (HEV) is a zoonotic pathogen and wild boars are recognised as reservoirs. HEV is the most common cause of acute viral hepatitis worldwide and it is usually transmitted by faecal-oral route. EU reported more than 21.000 human cases over the last 10 years (1). At the present, HEV comprises 8 genotypes and several subtypes. HEV genotypes 3 and 4 (HEV-3 and HEV-4) are zoonotic. This is the first report of HEV-3 subtype e and f in the wild boar population in Abruzzo region. Methods During the hunting season 2018-2019, liver samples from wild boars were analysed using Real Time RT-PCR for the detection of HEV RNA. Among these, positive sample were subjected to Whole Genome Sequencing for genotyping. After the quality check, reads were mapped using the nearest identified ref. sequence (KU176129) and the resulting HEV RNA sequences have been analysed using HEV net (open source software). Results Overall, 40 samples were tested positive for the detection of HEV RNA. Among these, 19 samples, with a Ct value ≤ 38, were selected for phylogenetic analysis. All RNA viral samples belonged to HEV-3. In particular, 2 RNA viral samples belonged to subtype e and 1 to subtype f. The others, instead, belonged to HEV-3 subtype c. Conclusions Up to now, only HEV-3 subtype c has been reported among wild boars in Abruzzo, an Italian region officially recognized as an HEV hot spot because of the high number of human cases and for the strong sero-prevalence among blood donors. In this study, we report for the first time the presence of HEV-3 subtypes e and f in the wild boar population of this area. Since HEV-3 subtypes e and f are commonly associated to human disease from consumption of raw/undercooked pork meats, these findings suggest the potential risk also associated with the consumption of wild boar meat. Nevertheless, this speculation needs further investigations. 1.EFSA (2017). Public health risks associated with HEV as a food-borne pathogen. E. Jou., 15(7) Key messages First report of the presence of HEV-3 subtypes e and f among wild boar population in Abruzzo region. Potential risk for human health of HEV transmission associated with raw/undercooked wild boar meat consumption.

INCIDENCE AND CIRCULATION OF HEPATITIS E VIRUS AMONG FARMED RABBITS IN EGYPT

The rabbit hepatitis E (rHEV) as been found in farmed, wild and pet rabbits as well as in human patients suggesting zoonotic transmission. The aim of this study was to evaluate the prevalence of HEV-infection in farmed rabbits from hyperendemic regions. Serum anti-HEV was tested by ELISA. We cоllected blоod samples from 235 rabbits from different governоrate in Egypt. HEV RNA was tested using RT-PCR with degenerative primers to ORF2 in serum and feces samples from 235 and 323 farmed rabbits respectively at different governorates in Egypt. All animals were 2-12 months of age. Out оf the 235 rabbit serum samples, the total positive rate of anti‐HEV IgG was 28/235 (11.9%). Anti-HEV IgG prevalence in rabbits was demonstrated by EIA in serum samples in different governorates 13.20%, 13.6 %, 14.6%, 24.2%, 20.0 % in Luxor, Assiut, Fayoum, Menoufia, and Alexandria respectively, and not detected in Sohag and Qena governorates. HEV RNA was detected in serum and fecal samples, only 2 serum samples were positive for HEV RNA 2/235 (0.8 %) at Luxor and Menoufia governorates. Out of the 323 fecal samples, the total positive rate HEV RNA was 26/323 (8.0%). Prevalence of HEV RNA in fecal samples of rabbits varied in different studied governorates from 1.92% to 18.36%, in Qena and Menoufia governorates respectively. HEV prevalence peaked in different age groups at different farms, with majority of infections at age low 6 months. Rabbit HEV infection in Egypt was first documented in our study.

Clinical significance of post-liver transplant hepatitis E seropositivity in high prevalence area of hepatitis E genotype 3: a prospective study

High hepatitis E (HEV) seroprevalence has been reported in the general population and in post-liver transplant (LT) cases in several regions, including Thailand, with genotype 3 being a predominant genotype. We hypothesized that HEV might persist at a subclinical level and might pose clinical risks in the post-LT period. We performed a cross-sectional study with 108 post-LT patients and found an IgG seroprevalence of 55.6%. Subsequently, 91 cases without clinical evidence of HEV-related hepatitis were enrolled in 1 year of prospective follow-up to determine clinical status, serologies and serum/feces HEV RNA every 4 months. HEV RNA was detected, indicating subclinical infections in patients with or without seropositivity, with an annual incidence of 7.7%. Our results suggest that subclinical HEV infection exists among LT patients in this high-prevalence area. Thus, clinicians should be aware of the possibility of disease reemergence and HEV viral transmission in LT patients.

Loss of Anti-Hepatitis E Virus Antibodies in Organ Transplant Patients with Hepatitis E

Background: To date, no reports exist regarding changes in anti-hepatitis E virus (HEV) antibodies after HEV infection in organ transplant patients. We followed up participants in a previously reported “Japanese national survey of Hepatitis E in liver, heart, and kidney transplant patients” from 2012 to 2017, to clarify HEV infection trends. Methods: Data on 106 transplant patients that were included in the current study; 32 liver, seven heart, and 67 kidney transplant recipients in 16 institutes in Japan were examined for immunoglobulin G (IgG), IgM, IgA classes of anti-HEV antibodies, and HEV RNA in the serum. Two, one, and five liver, heart, and kidney transplant patients, respectively, in whose serum HEV RNA was identified were examined. Furthermore, 30, six, and 62 liver, heart, and kidney transplant patients, respectively, positive for anti-IgG-HEV antibody were also examined. In each patient, HEV RNA, anti-HEV IgA antibody, anti-HEV IgM antibody, and anti-HEV IgG antibody titres were measured. Findings: Of eight patients with hepatitis E who were positive for HEV RNA, one (14·0%) had anti-IgG-HEV antibody negative conversion. Contrariwise, of 98 patients that were negative for HEV RNA, 24 (24·5%) had anti-HEV IgG antibody negative conversion. Of 24 patients, eight, 12, and four were liver, kidney, and heart transplant patients, respectively. Interpretation: Anti-HEV IgG antibody loss may occur unexpectedly with high frequency in transplanted patients with past HEV infection. Therefore, the possibility of HEV re-infection in transplanted patients exists, and care should be taken to prevent this. Funding: This research was supported by AMED (Grant Number JP19fk0210043). Declaration of Interests: None declared. Ethical Approval Statement: The date of birth, sex, date of transplantation, and number of immunosuppressant were extracted from the medical records. This research was approved by the Ethics Committee at our centre and at each patient's institution (IB1811). All the recipients gave written informed consent to participate in this study.

Four-year long (2014-2017) clinical and laboratory surveillance of hepatitis E virus infections using combined antibody, molecular, antigen and avidity detection methods: Increasing incidence and chronic HEV case in Hungary

BackgroundHepatitis E virus (HEV) is a pathogen of viral hepatitis. Since 2006, the number of reported HEV cases has ten-fold increase in Hungary. ObjectivesThe aim of this clinical and laboratory surveillance study was to analyse and confirm HEV IgM-positive sera with different methods in four consecutive years (2014–2017) in Hungary. Study designBetween 2014 and 2017, a total of 1439 sera samples were tested for HEV from in/out-patients with unknown hepatitis from university and county hospitals and general practitioners from three counties in Southwest Hungary (covered population: Σ894.000 persons) using combined antibody (serology), various molecular (RT-PCR and RT-qPCR), novel antigen (Ag) and avidity detection methods. ResultsTotal of 162 (11.3%) of the 1439 sera were HEV IgM-positive including 13 (8%) HEV RT-PCR-positive (confirmed as HEV genotype 3 sub-genotypes 3a/c/e/f/i in genus Orthohepevirus A) with up to 1.1383 × 108 RNA copy/ml, 30 (18.5%) HEV Ag-positive and 16 with low avidity index for HEV, respectively. Total of 6 samples were positive simultaneously with the combined four methods and 31 with three methods. If the quotient of serum sample's OD/cut-off of anti-HEV ELISA IgM and IgG scores is higher than ≥1 it predisposes for acute HEV infection. No rat or ferret HEV RNA (genus Orthohepevirus C) were identified from these specimens by RT-PCR. During our surveillance period a 68-year-old professional (meat-packing) hunter with kidney transplantation and immunosuppressive therapy was confirmed and treated as the first documented case of chronic HEV infection in Hungary. ConclusionThis four-year-long clinical and laboratory surveillance highlights the increasing importance of acute and chronic HEV infections in Hungary and supports the use of confirmatory assays for laboratory diagnosis of HEV in human.

Hepatitis E Genotype 1 Outbreak in Jharkhand, India: A Descriptive Analysis.

A waterborne outbreak of hepatitis E virus occurred in the Jamshedpur city of Jharkhand from March 2018 to October 2018. The aim of the present study is to study the hepatitis E virus outbreak clinically, serologically and etiologically. Five hundred and eighty-four clinically and biochemically documented cases were screened for the hepatotropic viral markers (HepatitisA, B, C, and E) by the ELISA. Demographic data such as gender, age, clinical diagnosis, location, outcome and time of admission were extracted from the online hospital management system. Water samples from the affected area were tested for HEV RNA detection. Genotyping of HEV virus was carried out by sequencing and phylogenetic analysis. Hepatitis E genotype 1 was confirmed as the major etiological agent in this outbreak due to the faecal contamination of drinking water supply while establishing illegal water connections. Mixed infection of HEV-HAV (5.31%) or HEV-HBV (0.91%) was also detected in the present series of acute viral hepatitis. The study highlights the importance of screening for both enterically transmitted hepatotropic viral markers as well as the parenterally transmitted hepatotropic viral markers during the outbreaks of acute viral hepatitis.