Detection Of Hepatitis E Virus Research Articles

Evaluation of Food Homogenates on Cell Survival In Vitro.

A critical review on the approaches to assess the infectivity of the Hepatitis E virus (HEV) in food recommended that a cell culture-based method should be developed. Due to the observations that viral loads in food may be low, it is important to maximise the potential for detection of HEV in a food source in order to fully assess infectivity. To do so, would require minimal processing of any target material. In order to proceed with the development of an infectivity culture method that is simple, robust and reproducible, there are a number of points to address; one being to assess if food homogenates are cytotoxic to HEV susceptible target cells. Food matrices previously shown to have detectable HEV nucleic acid were selected for analysis and assessed for their effect on the percentage survival of three cell lines commonly used for infectivity assays. Target cells used were A549, PLC/PRF/5 and HepG2 cells. The results showed that, as expected, various food homogenates have differing effects on cells in vitro. In this study, the most robust cell line over a time period was the A549 cell line in comparison to HepG2, with PLC/PRF/5 cells being the most sensitive. Overall, this data would suggest that FH can be left in contact with A549 cells for a period of up to 72h to maximise the potential for testing infection. Using food homogenates directly would negate any concerns over losing virus as a result of any additional processing steps.

High Prevalence of Hepatitis E IgM Antibody among Pregnant Women in Their Second and Third Trimester in Southwest Nigeria

Background: Background: Pregnant women with Hepatitis E virus (HEV) infection, particularly those in the second or third trimester, are at increased risk of acute liver failure, fetal loss, and mortality; but unfortunately, routine HEV screening is yet to be introduced for pregnant women in Nigeria and many developing nations. This study was conducted to determine HEV prevalence and the associated risk factors among pregnant women unaware of their HEV status in three states of the southwestern geopolitical zone in Nigeria. Methods: Two hundred and thirty-two consenting pregnant women aged 18 -55 years from Lagos, Oyo, and Osun state were recruited for the study and screened for HEV IgM antibodies using DIAPRO HEV IgM ELISA KIT (Milanno-Italy). Sociodemographic data and risk factors were obtained using a structured questionnaire, and data were analyzed using SPSS version 23. Results: An overall HEV prevalence rate of 9.9% was observed among pregnant women in Southwest Nigeria. State-specific prevalence of 14.7%, 8.5%, and 6.7% was observed in Oyo, Lagos, and Osun, respectively. HEV prevalence increased with increase in trimester of pregnancy, with more than 80% of the HEV prevalence found among women in their second and third trimester. All the HEV prevalence occurring in women of active childbearing age (≤ 40 years) and among low-income earners. None of the risk factors considered was found to have a statistical correlation with HEV prevalence. Conclusion: High HEV prevalence in the second and third trimesters was reported in southwest Nigeria. We advocate routine screening of pregnant women for HEV IgM antibody for prompt HEV detection and proper HEV management to forestall HEV complications in pregnancy.

Assessing the efficacy of different bead-based assays in capturing hepatitis E virus

Hepatitis E virus (HEV) generally causes acute liver infection in humans and its transmission could be waterborne, foodborne, bloodborne, or zoonotic. To date, there is no standard method for the detection of HEV from food and environmental samples. Herein, we explored the possibility of using magnetic beads for the capture and detection of HEV. For this purpose, we employed Dynabeads M-270 Epoxy magnetic beads, coated with different monoclonal antibodies (mAbs) against HEV capsid protein, and the Nanotrap Microbiome A Particle magnetic beads, which are coated with chemical affinity baits, to capture HEV-3 particles in suspension. Viral RNA was extracted by heat-shock or QIAamp viral RNA kit and subjected to quantification using digital-droplet RT-PCR (ddRT-PCR). We demonstrated that the mAb-coupled Dynabeads and the Nanotrap particles, both were able to successfully capture HEV-3. The latter, however had lower limit of detection (<140gc compared with <1400 gc) and significantly higher extraction efficiency in comparison to the mAb-coupled Dynabeads (41.1% vs 8.8%). We have also observed that viral RNA extraction by heat-shock is less efficient compared to using highly denaturing reagents in QIAmp viral RNA extraction kit. As such, magnetic beads have the potential to be used to capture HEV virions for research and surveillance purposes.

Development and clinical validation of a one-step pentaplex real-time reverse transcription PCR assay for detection of hepatitis virus B, C, E, Treponema pallidum, and a human housekeeping gene

BackgroundWith the safety of blood transfusion being a major public health concern, the development of a rapid, sensitive, specific, and cost-effective multiplex PCR assay for simultaneous detection of hepatitis B virus(HBV), hepatitis C virus (HCV), hepatitis E virus (HEV), and Treponema pallidum(T. pallidum) in blood is crucial.MethodsFive primer pairs and probes were designed towards conserved regions of target genes and used to establish a one-step pentaplex real-time reverse transcription PCR(qRT-PCR) assay for simultaneous detection of HBV, HCV, HEV, T. pallidum, and RNase P(housekeeping gene), providing sample quality check. The clinical performance of the assay was further determined with 2400 blood samples from blood donors and patients in Zhejiang province, and compared the results with commercial singleplex qPCR and serological assays.ResultsThe 95% limit of detection(LOD) of HBV, HCV, HEV, and T. pallidum were 7.11 copies/µL, 7.65 copies/µL, 8.45 copies/µL, and 9.06 copies/µL, respectively. Moreover, the assay has good specificity and precision. Compared to the singleplex qPCR assay, the novel assay for detecting HBV, HCV, HEV, and T. pallidum presented 100% clinical sensitivity, specificity, and consistency. Several discrepant results between serological and pentaplex qRT-PCR assays were found. Of 2400 blood samples, there were 2(0.08%) HBsAg positive samples, 3(0.13%) anti-HCV positive samples, 29(1.21%) IgM anti-HEV positive samples and 6(0.25%) anti-T. pallidum positive samples proven negative in nucleic acid detection. 1(0.04%) HBV DNA positive sample and 1(0.04%) HEV RNA positive sample were detected negative by serological testing.ConclusionsThe developed pentaplex qRT-PCR is the first assay on simultaneous, sensitive, specific, and reproducible detection of HBV, HCV, HEV, T. pallidum, and RNase P in a single tube. It could detect pathogens in blood during the window period of infection and is a good tool for effectively screening blood donors and early clinical diagnosis.

Preliminary investigation of hepatitis E virus detection by a recombinase polymerase amplification assay combined with a lateral flow strip.

Hepatitis E virus (HEV) is a zoonotic pathogen that is a significant public health problem. Detecting HEV relies mainly on conventional PCR, which is time-consuming and requires sophisticated instruments and trained staff. We aimed to establish a reverse-transcription (RT)-recombinase polymerase amplification (RPA) assay (RT-RPA) combined with a lateral flow strip (LFS; RT-RPA-LFS) to rapidly detect HEV RNA in human and rabbit samples. With the optimal reaction conditions (37°C for 30 min), our assay detected as few as 1.0 × 102 copies/mL of HEV and showed no cross-reactivity with other hepatitis viruses. We tested 28 human samples (4 fecal and 24 serum samples) and 360 rabbit samples (180 fecal and 180 serum samples) with our RT-RPA-LFS assay and compared our assay to an RT-qPCR method. There was no significant difference (p > 0.05) in the test results between the 2 assays. Our RT-RPA-LFS assay detected both HEV3 and HEV4 genotypes. Our rapid, sensitive, and specific RT-RPA-LFS assay for the detection of HEV may provide a useful detection tool for limited-resource areas.

Hepatitis E Virus (HEV) Spreads from Pigs and Sheep in Mongolia

Hepatitis E is a viral infectious disease in pigs, wild boars, cows, deer, rabbits, camels, and humans as hosts caused by Paslahepevirus. Recently, it has been detected in a wide variety of animals including domestic small ruminants. Mongolia is a land of nomadic people living with livestock such as sheep, goats, and cattle. Due to how Mongolian lifestyles have changed, pork has become popular and swine diseases have emerged. Among them, Hepatitis E disease has become a zoonotic infectious disease that needs to be addressed. The HEV problem in pigs is that infected pigs excrete the virus without showing clinical symptoms and it spreads into the environment. We attempted to detect HEV RNA in sheep which had been raised in Mongolia for a long time, and those animals living together with pigs in the same region currently. We also conducted a longitudinal analysis of HEV infection in pigs in the same area and found that they were infected with HEV of the same genotype and cluster. In this study, we examined 400 feces and 120 livers (pigs and sheep) by RT-PCR in Töv Province, Mongolia. HEV detection in fecal samples was 2% (4/200) in sheep and 15% (30/200) in pigs. The results of ORF2 sequence analysis of the HEV RT-PCR-positive pigs and sheep confirmed genotype 4 in both animals. The results suggest that HEV infection is widespread in both pigs and sheep and that urgent measures to prevent infection are needed. This case study points to the changing nature of infectious diseases associated with livestock farming. It will be necessary to reconsider livestock husbandry and public health issues based on these cases.

Label-free ultra-sensitive colorimetric detection of hepatitis E virus based on oxidase-like activity of MnO2 nanosheets.

Hepatitis E virus (HEV) is an evolving infectious entity that causes viral hepatitis infections worldwide. Current routine methods of identifying and diagnosing HEV are someway laborious and costly. Based on the biomimicking oxidase-like activity of MnO2 nanosheets, we designed a label-free, highly sensitive colorimetric sensing technique for HEV detection. The prepared MnO2 catalyst displays intrinsic biomimicking oxidase-like catalytic activity and efficiently oxidizes the 3,3',5,5'-tetramethylbenzidine (TMB) substrate from colorless to blue colored oxidized TMB (oxTMB) product which can be measured at 652nm by UV-visible spectrum. When the HEV-DNA was added, DNA adsorbed easily on MnO2 surface through physical adsorption and electrostatic interaction which hinders the oxidase-like catalytic activity of MnO2. Upon the introduction of target, the HEV target DNA binds with its complementary ssDNA on the surface of MnO2, the hybridized DNA releases from the surface of MnO2, which leads to recovery of oxidase-like catalytic activity of MnO2. This strategy was applied to construct a colorimetric technique for HEV detection. The approach works in the linear range of 1 fM-100nM DNA concentration with the limit of detection (LOD) of 3.26 fM (S/N = 3) and quantitative limit (LOQ) of 36.08 fM. The TMB-MnO2 platform was highly selective for HEV target DNA detection when compared with potential interferences. Result of serum sample analysis demonstrates that this sensing system can be used for clinical diagnostic applications.

Rapid and direct detection of hepatitis E virus in raw pork livers by recombinase polymerase amplification assays.

Hepatitis E virus (HEV) is a zoonotic pathogen that causes global hepatitis E. Outbreaks of hepatitis E are directly linked to the consumption of pork liver products. Herein reverse transcription recombinase polymerase amplification assays targeting the ORF2 gene were developed for the rapid detection of HEV by integrating the fluorescence detection platform (qRT-RPA) and the visible lateral flow biosensor by naked eyes (LFB RT-RPA). The qRT-RPA assay effectively detected HEV RNA with a limit of detection (LOD) of 154 copies/μl (95%CI: 126–333 copies/µl) in Genie III at 41°C for 20 min. Besides this, the LFB RT-RPA detected the HEV RNA with a LOD of 24 copies/μl (95%CI: 20–57 copies/µl) in an incubator block at 41°C for 20 min. The developed RT-RPA assays also showed good specificity for HEV, with no cross-reactions with any of the other important swine pathogens examined in this work. The performance of the developed RT-RPA assays was validated on 14 HEV RNA-positive and 66 HEV RNA-negative raw pork liver samples identified by a previously described qRT-PCR. Consequently, 11 and 12 samples were HEV RNA-positive as detected by the qRT-RPA and the LFB RT-RPA, respectively. Compared to qRT-PCR, the qRT-RPA and LFB RT- RPA assays revealed a coincidence rate of 96.3 and 97.5% as well as a Kappa value of 0.858 and 0.908, respectively. These results ascertain that the developed RT-RPA assays are effective diagnostic tools for the point-of-care detection of HEV in resource-limited settings.

Detection of hepatitis E virus in milk: Current evidence for viral excretion in a wide range of mammalian hosts.

Infection with hepatitis E virus (HEV) is common in both developing and industrialized nations. Genotypes 3 and 4 are increasingly being reported, particularly in high-income countries where the precise extent of HEV transmission via food is currently unclear. Recently, HEV has been found to be excreted in milk; however, data on the potential milk-borne transmission is still lacking or conflicting and warrants further research on the topic. As such, the aim of the present study was to review the current scientific knowledge and to summarize the existing studies in which HEV has been detected in milk. Exhaustive searches were carried out in Mendeley, PubMed, Scopus and Web of Science. A total of 157 papers were retrieved from the four electronic databases. After removing duplicate articles from the databases (n = 30), exclusion criteria identified unrelated research (n = 115). This allowed the identification of 12 eligible papers. To date, studies on HEV detection in milk were mostly from China (n = 5), followed by Egypt (n = 2), Germany (n = 1), Belgium and Holland (n = 1), Turkey (n = 1), Czech Republic (n = 1) and Spain (n = 1) and were focused on a variety of animals (cow, goat, donkey, buffalo, sheep and camel) and humans. Four out of the 12 eligible studies did not find any evidence of HEV in milk. Moreover, 3 out of the 12 studies detected low rates of HEV (0.2-1.8%) and two were based on a low sample size (n = 1 and n = 4). Interestingly, one study showed very high detection rates and also detected HEV genotype 1 in an animal milk sample, an unusual finding since it only occurs in humans, deserving further studies for confirmation and characterization. Two studies detected high prevalence of HEV genotype 4 in bovine samples from China, with one showing indication of the presence of infectious HEV in milk. To date, there is still a small amount of available data on the HEV presence in milk, posing important questions regarding both animal and human health. Thus, further efforts on this potentially underestimated zoonotic route for HEV should be given, warranting further studies on the topic.

Real-Time PCR-Based Methods for Detection of Hepatitis E Virus in Pork Products: A Critical Review.

Standard methods for detection of hepatitis A virus and norovirus in at-risk foodstuffs are available, but currently there is no standard method for detection of hepatitis E virus (HEV) in pork products or other foods that can be contaminated with the virus. Detection assays for HEV are mainly based on nucleic acid amplification, particularly the reverse transcription polymerase chain reaction (RTPCR) in real-time format. RTPCR-based methods can be sensitive and specific, but they require a suite of controls to verify that they have performed correctly. There have been several RTPCR methods developed to detect HEV in pork products, varying in details of sample preparation and RTPCR target sequences. This review critically discusses published HEV detection methods, with emphasis on those that have been successfully used in subsequent studies and surveys. RTPCR assays have been used both qualitatively and quantitatively, although in the latter case the data acquired are only reliable if appropriate assay calibration has been performed. One particular RTPCR assay appears to be ideal for incorporation in a standard method, as it has been demonstrated to be highly specific and sensitive, and an appropriate control and calibration standard is available. The review focuses on the detection of HEV in pork products and similar foodstuffs (e.g., boar). The information may be useful to inform standardisation activities.

Nationwide survey of hepatitis E virus infection among wildlife in Japan

In Japan, hepatitis E virus (HEV) causes hepatitis in humans through the consumption of raw or undercooked meat, including game meat. In the present study, nationwide surveillance of HEV infection among a total of 5,557 wild animals, including 15 species, was conducted in Japan. The prevalence of anti-HEV antibodies in wild boar was 12.4%, with higher positive rates in big boars (over 50 kg, 18.4%) than in small individuals (less than 30 kg, 5.3%). Furthermore, HEV RNA was more frequently detected in piglets than in older boars. Interestingly, the detection of HEV among wildlife by ELISA and RT-PCR suggested that HEV infection in Sika deer was a very rare event, and that there was no HEV infection among wild animals except for wild boar, Sika deer and Japanese monkeys. In conclusion, wild boar, especially piglets, are at high risk of HEV infection, while other wild animals showed less risk or no risk of HEV transmission.

Detection of hepatitis E virus RNA from pig bile collected at a slaughterhouse in Japan.

Hepatitis E virus (HEV) is a zoonotic pathogen that circulates mainly between pigs and humans. In Japan, the number of confirmed HEV cases has increased over the past decade, with the majority reported as domestic HEV infections. HEV-infected pork products may be associated with this increase, but there is limited information on HEV in pork in Japanese markets. From February to March 2020, gallbladders were collected from 200 slaughtered pigs shipped from 14 farms and were surveyed to detect HEV RNA in bile using reverse transcription quantitative polymerase chain reaction. The samples were then sequenced and genotyped. Twenty pigs were positive for HEV ribonucleic acid, and seven samples had Ct values of less than 30. Among these 20 pigs, virus strains from 14 pigs were determined as genotype 3. This report indicated that HEV-contaminated pork liver was shipped to consumer markets and demonstrated the importance of detection of HEV in meat ready for shipment.

Isolation, identification and genome analysis of an avian hepatitis E virus from white-feathered broilers in China

Avian hepatitis E virus (HEV) is the major causative pathogen of the big liver and spleen disease, hepatitis-splenomegaly syndrome, and hepatic rupture hemorrhage syndrome. Until now, there are 6 different avian HEV genotypes that infect chickens have been reported worldwide. Epidemiologic investigations of the avian HEV demonstrated that avian HEV has been widely spread in China in recent years. In this study, an avian HEV named YT-aHEV was obtained from white-feathered broilers using LMH cells by virus isolation assay in Shandong province, China. The complete genome consists of 6656-nt excluding the poly(A) tail. The isolate was highly similar to the CaHEV strain and segregated into the same branch belonging to avian HEV genotype 3. Indirect immunofluorescence using capsid protein-specific polyclonal antibodies confirmed that YT-aHEV could establish productive infection and replicate stably in LMH cells. Furthermore, an in vivo avian HEV infection model was established successfully in specific pathogen-free chicken embryos by intravenous experiments. In the present study, we demonstrate an avian HEV infection associated with liver lesions of hemorrhage and swelling by LMH cells for the first time in a white-feather broiler flock in China. This research also provides a new diagnosis method for detection of avian HEV, which laid a foundation for the understanding of pathogenicity and molecular biology of this virus for further study.

First molecular detection of hepatitis E virus genome in camel and pig faecal samples in Ethiopia

BackgroundHepatitis E is an enteric and zoonotic disease caused by hepatitis E virus (HEV) that is mainly transmitted via the faecal-oral route through contaminated food or the environment. The virus is an emerging infectious agent causing acute human infection worldwide. A high seroprevalence of the disease was reported in pregnant women in Addis Ababa, Ethiopia, raising significant public health concern. The presence of HEV specific antibodies were also reported in dromedary camels in the country; however, the infectious virus and/or the viral genome have not been demonstrated to date in animal samples.MethodsTo address this gap, a total of 95 faecal samples collected from both apparently healthy pigs of uncharacterised types (50 samples) in Burayu and Addis Ababa areas and camels (Camelus dromedarius, 45 samples) in west Hararghe were screened for the presence of HEV genome using universal primers in a fully nested reverse transcription polymerase chain reaction (nRT-PCR). The protocol is capable of detecting HEV in faecal samples from both pigs and camels.ResultsThe nRT-PCR detected HEV genes in six (12%) pig faecal samples and one camel sample (2.2%). Therefore, the results indicate that HEV is circulating in both pigs and camels in Ethiopia and these animals and their products could serve as a potential source of infection for humans.ConclusionThe detection of HEV in both animals could raise another concern regarding its public health importance as both animals’ meat and camel milk are consumed in the country. Further studies to determine the prevalence and distribution of the virus in different animals and their products, water bodies, food chain, and vegetables are warranted, along with viral gene sequencing for detailed genetic characterisation of the isolates circulating in the country. This information is critically important to design and institute appropriate control and/or preventive measures.

Significant compartment-specific impact of different RNA extraction methods and PCR assays on the sensitivity of hepatitis E virus detection.

RNA detection in plasma/stool is the gold-standard for diagnosis of hepatitis E virus (HEV) infection. The impact of viral extraction methods on HEV RNA detection is poorly investigated. We determined the limit of detection of the RealStar HEV RT-PCR V2.0 Kit (altona Diagnostics, RS) utilizing 3 RNA extraction methods (COBAS® AmpliPrep Total Nucleic Acid Isolation Kit, TNAi Roche; MagNA Pure 96 DNA, Viral NA SV Kit, MgP; QIAamp Viral RNA mini Kit Qiagen; VRK) in plasma and stool. The most sensitive method was evaluated in a total of 307 longitudinal samples of patients with HEV infection (acute=18/chronic=36) and compared to results with the former diagnostic standard of our centre (TNAi/FastTrack Diagnostic; FTD). The plasma-LOD was 49, 94 and 329IU/mL for extraction with MgP, VRK and TNAi respectively. In stool, the LOD was 21IU/mL, 528IU/mL and indefinable for extraction with TNAi, VRK and MgP respectively. Utilizing longitudinal patient plasma samples, MgP/RS revealed 56 HEV RNA-positive samples in 158 negative samples as determined by TNAi/FTD. In stool, from 37 HEV negative samples (TNAi/FTD), 15 were positive with TNAi/RS. At end of treatment, 8 out of 27 chronically infected patients were RNA positive with MgP/RS, while classified negative with TNAi/FTD. A relapse occurred in 3 of these patients. Different methods for RNA extraction and quantification have a significant, compartment-specific impact on the sensitivity of HEV detection. Knowledge about the favourable combinations of extraction and quantification has important implications for diagnosis and patients receiving antiviral therapy.

Optimization and Implementation of the Virus Extraction Method for Hepatitis E Virus Detection from Raw Pork Liver.

Hepatitis E virus (HEV) has been frequently detected from pork liver and liver products, which can usually cause self-limiting diseases in healthy adults, yet may result in fatality in immunosuppressed groups. Nevertheless, there is so far no standardized method for HEV detection available from pork liver and/or liver products. The present study aimed to optimize the virus extraction method of HEV from raw pork liver, which is often consumed in Asia undercooked to avoid a grainy texture. By comparing different sample preparation protocols and by applying the selected protocol to 60 samples collected from Singapore retail markets, we demonstrated that homogenization of 0.25g raw pork liver with FastPrep™ Lysing Matrix Y containing yttria-stabilized zircondium oxide beads in 2ml tubes and with harsh mechanical force at 6ms-1, 40s/cycle, for 5 cycles with 300s pause time after each cycle is promising in both releasing the potentially intracellular viruses and resulting in satisfactory virus recovery rates (> 1%). A high prevalence (52%) of HEV genome was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) from the 60 samples collected from Singapore retail markets imported from Indonesia, Australia and Malaysia. However, RNase treatment decreased the HEV prevalence to 33.3%, and all of the 20 positive samples were with high RT-qPCR Ct values above 35, suggesting that the positive RT-qPCR signals maybe largely due to the inactive viruses and/or exposed HEV RNA traces in raw pork liver products. Therefore, conscious care should be taken when interpreting molecular detection results of viruses from food samples to be correlated with public health risks.

Prevalence of HAV and HEV in Patients Presenting with Acute Viral Hepatitis in Silchar Medical College & Hospital

Background: Hepatitis A virus (HAV) and Hepatitis E virus (HEV) are both enterically transmitted, resulting in acute viral hepatitis (AVH) in developing countries. Both are self-limiting viral infection, transmitted by feco-oral route. Exposure rate of HAV and HEV over a period are different in different parts of country. Aim: This study was done to determine prevalence of HAV and HEV in patients presenting with AVH and the co-infection of HAV and HEV in these patients. Materials and Methods: A cross sectional study of 2 years duration was conducted in the Department of Microbiology, SMCH, Silchar. A non random sampling of 200 patients presenting with AVH was considered in the study. On the basis of history, serum sample were analyzed for IgM anti HAV and IgM anti HEV for the detection of HAV and HEV, respectively using commercially available ELISA kits. Results: The prevalence of HAV and HEV co-infection in patients with acute viral hepatitis was 32.5% (65 cases). HAV and HEV were most prevalent between 20 to 35years age group. Hepatitis A samples only 60 were positive and Hepatitis E samples only 5 were positive. Conclusion: Almost all HAV and HEV infectious are spread by feco-oral route. The best means of reducing the spread of infection is by promoting simple measures of personal and community hygiene such as hand washing before eating and after defecation, the sanitary disposal of excreta which will prevent contamination of water, food and milk, during epidemics, boiled water should be advocated for drinking purposes.

First Report of Hepatitis E Virus in Shellfish in Southeast Italy

Hepatitis E virus (HEV) represents one of the principal causative agents of hepatitis globally. Among the five HEV genotypes affecting humans, genotypes 3 and 4 are zoonotic and are the main source of hepatitis E in developed countries. HEV has been detected in several foods. The present work investigated the presence of this virus in shellfish sold at retail in the Apulia region of Italy. The presence of HEV RNA was assessed by real-time RT-PCR in 225 shellfish samples collected during 2018. Overall, two (0.89%) of these samples tested positive for HEV RNA. To our knowledge, this is the first notification of the detection of HEV in mussels sold at retail in the Apulia region. These data highlight the potential role of shellfish as a vehicle for the transmission of viral pathogens.

Presence of hepatitis E virus in commercially available pork products

An increasing number of hepatitis E virus (HEV) infections in industrialized countries have been foodborne and linked to the consumption of undercooked pork products. To date, data on the prevalence of HEV in pork products sold in the United States is limited and no standard processing method exists for the detection of HEV in foods. In order to develop a processing method for the detection of HEV in pork products, ground pork and pork liver were selected for method development. Murine norovirus (MNV) was used as a process control. A filtration step prior to RNA detection was shown to reduce the level of PCR inhibitors in ground pork and an additional ultracentrifugation process was successful in removing PCR inhibitors in pork liver. MNV RNA was detected in ground pork and liver samples inoculated with 4.7 log10 PFU/g and 3.0 log10 PFU/g, respectively. Using the developed method for viral RNA detection in ground pork and pork liver, 20 packages of ground pork (six 1 g sub-samples per package) and 14 pork livers (four 1 g sub-samples per liver) were screened for the presence of HEV RNA. Fifteen out of 119 (12.6%) ground pork samples tested positive for HEV RNA and 13 out of 20 packages (65%) contained at least one positive sample. Twenty-five of 56 (45%) of pork liver samples were positive for HEV RNA and 6 of 14 livers (43%) had all sub-samples test positive for HEV RNA. Overall, the results indicate ground pork and pig liver as a potential source of HEV.

Infection for the Hepatitis E virus in pregnant women of Córdoba, Argentina

Background: The hepatitis E virus (HEV) is an important and poorly studied pathogen of worldwide distribution that causes enteric hepatitis. The HEV infection in humans is usually self-limited. However, cases of severe fulminant hepatitis have been reported in pregnant women and immunosuppressed patients. In Argentina, serological evidence of human HEV infection has been described in different geographical regions. In Córdoba, HEV prevalence of 4.4% in general population, 7.3% in HIV + individuals, 5.8% in solid organ transplant patients and 10.2% in hemodialysis patients have been documented. Besides, RNA-HEV detections have been performed and genotype 3 was detected. The aim of this research was to describe the circulation of HEV in pregnant women from Córdoba, know if there were symptomatic cases, analyse risk factors related to the infection, and compare the findings with the non-pregnant control group Methods and materials: A total of 202 serum samples from pregnant women (52 prospective and 150 retrospective samples), and 155 samples from non-pregnant women with the same ages (control group) from Córdoba were analysed for IgG anti-HEV detection. Positive samples were tested for IgM anti-HEV and RNA-HEV. Two samples from pregnant women with acute hepatitis were also analysed for HEV detection. Results: The seropositivity of IgG anti-HEV obtained for pregnant women was 8.4%, higher than the 2.6% obtained in the control group, and association between pregnancy and HEV infection was found (p = 0.023). There were no differences between seropositivity rates according to the trimester of pregnancy, although differences were found among age groups, with the higher prevalence in pregnant women younger than 25 years (16.7%, p = 0.032). From the IgG anti-HEV + patients, 2 presented IgM anti-HEV, although both were asymptomatic. None of these samples were positive for RNA-HEV detection. Conclusion: The present study shows, for the first time, the circulation of HEV in pregnant women from Córdoba, generating data that contributes to improve the surveillance and diagnosis of a virus little known in our environment, and under-diagnosed. These association between pregnancy and HEV infection should alert the local health team to consider this microorganism as an agent of liver disease in pregnant women.