Detección de anticuerpos anti virus hepatitis E IgG e IgM utilizando un método automatizado: experiencia de un hospital universitario / Detection of anti-virus hepatitis e IgG and IgM antibodies using an automated method: experience of a university hospital

Resumen La determinación de anticuerpos anti virus hepatitis E (anti-VHE) tiene gran variabilidad dependiendo del ensayo utilizado. En 2015, con un método ELISA manual, publicamos una seroprevalencia de anti-VHE IgG de 32,6% en pacientes con estudio de hepatitis. Existen escasas publicaciones de anti-VHE IgM. Recientemente, se desarrolló el primer método automatizado y en el presente estudio comunicamos la experiencia obtenida. Se analizaron los resultados de 272 pacientes con estudio de anti-VHE IgG y/o IgM mediante técnica automatizada ELFA (VIDAS®), entre mayo de 2018 y agosto de 2020. Se encontró 25,8% (68/264) de positividad para anti-VHE IgG y 3,5% (9/259) para anti-VHE IgM. Cuatro muestras tuvieron ambos anticuerpos positivos. La seropositividad de anti-VHE IgG aumentó con la edad. En conclusión, la seroprevalencia de anti-VHE IgG obtenida fue similar a la publicada previamente. Considerando las ventajas de los ensayos IgM e IgG anti-VHE en el sistema VIDAS®, parecen ser nuevas herramientas valiosas en el estudio serológico de VHE.

Abstract The determination of anti-hepatitis E virus antibodies (anti-HEV) has a high variability depending on the assay used. In 2015, with a manual ELISA method, we reported anti-HEV IgG seroprevalence of 32.6% in patients under hepatitis study. There are few reports of anti-HEV IgM. Recently, it was developed the first automated method and in the present study, we report the experience using this new method. Between May 2018 and August 2020, the results of 272 patients with an anti-HEV IgG and/or IgM study were analyzed using the automated ELFA technique (VIDAS®). Seroprevalence was 25.8% (68/264) for anti-HEV IgG and 3.5% (9/259) for anti-HEV IgM. Four samples were positive for both antibodies. Anti-HEV IgG seropositivity increased with age. In conclusion, the seroprevalence of anti-HEV IgG obtained was similar to previously reported. Taking into account the advantages of these assays, anti-HEV IgM and IgG assays on VIDAS® system, seem to be valuable new tools in serological study of HEV.

Hepatitis A and hepatitis E virus co-infection with right pleural effusion, ascites and acute acalculous cholecystitis. A case report / Coinfección por el virus de la hepatitis A y la hepatitis E con derrame pleural derecho, ascitis y colecistitis acalculosa aguda. Reporte de caso

ABSTRACT Hepatitis A and hepatitis E are the leading causes of acute viral hepatitis in developing countries due to our poor sanitary conditions, both spread by fecal-oral route or through contaminated water and food. Being both self-limiting diseases, they are usually benign but may present with atypical clinical findings. A 32 year-old female with right pleural effusion, ascites and acalculous cholecystitis during the course of HAV and HEV co-infection is reported. Clinical improvement was observed with conservative management. As far as we know, this is the first case described of a patient with these three complications in the background of a hepatitis A virus and hepatitis E virus co-infection.

RESUMEN Hepatits A y hepatitis E son las principales causas de hepatitis viral en países en desarrollo debido a las limitadas condiciones sanitarias. Son condiciones usualmente benignas y autolimitadas, pero pueden presentarse de forma atípica. Se reporta una paciente de 32 años con efusión pleural derecha, colecistitis acalculosa y ascitis en el curso de una co-infección por el virus de Hepatitis A y hepatitis E. Hasta donde tenemos conocimiento, este es el primer caso de una paciente con estas tres complicaciones como resultado de una infección por el virus de hepatitis A y hepatitis E.

Serological and molecular retrospective analysis of hepatitis E suspected cases from the Eastern Brazilian Amazon 1993-2014

Abstract INTRODUCTION We evaluated the anti-hepatitis E virus (HEV) antibody prevalence and HEV-RNA in archived serum samples of non-A-C hepatitis, or suspected cases of HEV infection from the Eastern Brazilian Amazon from 1993 to 2014. METHODS Serum samples (n = 318) were tested using ELISA and immunoblotting, and screened for HEV-RNA by RT-qPCR. RESULTS Anti-HEV IgM and IgG were detected in 3.4% (11/318) and 5.9% (19/318) of the samples, respectively. All samples were HEV-RNA negative. CONCLUSIONS HEV was detected at a low prevalence. Broader serological and molecular evaluation of HEV infection in the Amazon region should be carried out.

Hepatitis E virus infection among rural Afro-descendant communities from the eastern Brazilian Amazon

Abstract INTRODUCTION The prevalence, public health impact, and epidemiological characteristics of hepatitis E virus (HEV) are poorly understood in Brazil. METHODS Serum samples from 535 individuals from three rural Afro-descendant communities located in eastern Brazilian Amazon were collected in October 2015 and tested for presence of anti-HEV IgM and IgG antibodies. Serologically positive samples were also tested for HEV-RNA. RESULTS Two cases were confirmed for anti-HEV IgM (0.3 %) and two cases for anti-HEV IgG (0.3 %). No sample was positive for HEV-RNA. CONCLUSIONS Results indicated low prevalence of HEV infection in Afro-descendant rural communities from the eastern Brazilian Amazon.

Seroprevalencia de virus hepatitis E en donantes de sangre en un hospital universitario en Chile / Hepatitis E virus seroprevalence in blood donors in a university hospital in Chile

Resumen En Chile, existen escasos estudios de seroprevalencia de anticuerpos IgG anti virus hepatitis E (VHE) en bancos de sangre, entre 4 y 8%. El desarrollo de nuevas técnicas con mayor sensibilidad y especificidad, dan cuenta de un aumento de la seroprevalencia de VHE en diversos países, siendo desconocido el estado actual en Chile. En el presente estudio, determinamos la seroprevalencia de IgG anti VHE en donantes de sangre del Hospital Clínico Universidad de Chile, con técnicas de ELISA de última generación. De un total de 186 muestras, recolectadas el año 2014, 56 (30,1%) resultaron positivas, sin diferencias de género, pero con un incremento significativo con la edad (p < 0,001). Estos resultados muestran un aumento en la seroprevalencia de VHE en donantes de sangre realizados con inmunoensayos de mayor sensibilidad.

In Chile, there are few studies about seroprevalence of IgG antibodies against hepatitis E virus (HEV) in blood banks, between 4 and 8%. The development of new techniques with greater sensitivity and specificity, account for an increase in the seroprevalence of HEV in various countries, the current status in Chile being unknown. In the present study, we determined the seroprevalence of anti-HEV IgG in blood donors of the Clinical Hospital University of Chile, with last generation ELISA techniques. Out of a total of 186 samples, collected in 2014, 56 (30.1%) were positive, without gender differences, but with a significant increase with age (p < 0.001). These results show an increase in the seroprevalence of HEV in blood donors performed with immunoassays of greater sensitivity.

High prevalence of hepatitis E virus antibodies in Sao Paulo, Southeastern Brazil: analysis of a group of blood donors representative of the general population

Abstract Brazil is a non-endemic country for hepatitis E virus (HEV) infection with seroprevalence from 1% to 4% in blood donors and the general population. However, data on seroprevalence of HEV in the country are still limited. This study evaluated the prevalence of past or present HEV infection in a group of blood donors representative of the general population of the city of Sao Paulo, Southeastern Brazil. Serum samples from 500 blood donors were tested from July to September 2014 by serological and molecular methods. Anti-HEV IgG antibodies were detected in 49 (9.8%) subjects and categorized age groups revealed an age-dependent increase of HEV seroprevalence. Among the anti-HEV IgG positive subjects, only 1 had anti-HEV IgM while none tested positive for HEV-RNA. The present data demonstrate a higher seroprevalence of anti-HEV IgG than previously reported in the region.

Acute Hepatitis E Mimicking a Flare of Disease in a Patient with Chronic Autoimmune Hepatitis

Abstract: Acute hepatitis E is becoming increasingly recognised in Europe with up to 40% of the population in Southern France being exposed to the virus, which is harboured in pigs. Patients with known liver disease may present with acute hepatitis E and present a diagnostic challenge. For example patients with autoimmune hepatitis (AIH) who are immunosuppressed and contract hepatitis E may be at increased risk of developing chronicity due to concurrent immunosuppression. Importantly, the diagnosis may be missed with the infection misdiagnosed as an autoimmune flare, and immunosuppression increased by the attending physician, thus enhancing the risk of chronicity of infection leading to progressive liver injury in immunocompromised patients. We report a case of acute hepatitis E in a patient with AIH and discuss the features that helped us differentiating it from an autoimmune flare.

High Clinical Manifestation Rate in an Imported Outbreak of Hepatitis E Genotype 1 Infection in a German Group of Travellers Returning from India

Abstract: Background. There are only few reports about travel-associated, imported tropical hepatitis E virus (HEV) genotype 1 infections within Western travellers. We describe the clinical course of a single outbreak of hepatitis E in a German travellers group returning from India and compare the results of two commercial HEV-seroassays. Material and methods. After identifying hepatitis E in an index patient returning from a journey to India all 24 members of this journey were tested for anti-HEV-IgG and IgM using two commercial seroassays (Wantai and Mikrogen), for HEV-RNA by PCR and HEV-Ag by an antigen-assay (Wantai). Results. 5/24 (21%) individuals were viraemic with viral loads between 580-4,800,000 IU/mL. Bilirubin and ALT levels in these patients ranged from 1.3-14.9 mg/dL (mean 7.3 mg/dL, SD 5.6 mg/dL) and 151-4,820 U/L (mean 1,832U/L, SD 1842U/L), respectively and showed significant correlations with viral loads (r = 0.863, p < 0.001; r = 0.890, p < 0.001). No risk factor for food-borne HEV-transmission was identified. All viraemic patients (5/5) tested positive for anti-HEV-IgG and IgM in the Wantai-assay but only 4/5 in the Mikrogen-assay. Wantai-HEV-antigen-assay was negative in all patients. Six months later all previously viraemic patients tested positive for anti-HEV-IgG and negative for IgM in both assays. However, two non-viremic individuals who initially tested Wantai-IgM-positive stayed positive indicating false positive results. Conclusions. Despite the exact number of exposed individuals could not be determined HEV genotype 1 infections have a high manifestation rate of more than 20%.The Wantai-antigen-test failed, the Wantai-IgMrapid-test and the Mikrogen-IgM-recomblot showed a better performance but still they cannot replace real-time PCR for diagnosing ongoing HEV-infections.

Hepatitis E virus seroprevalence among schistosomiasis patients in Northeastern Brazil

Abstract Background Hepatitis E virus (HEV) can cause chronic infection with rapid progression to liver cirrhosis in immunocompromised patients. HEV seroprevalence in patients with Schistosoma mansoni in Brazil is unknown. We evaluated the prevalence of past or present HEV infection in schistosomiasis patients in Recife, Pernambuco, Brazil. A total of 80 patients with Schistosoma mansoni were consecutively enrolled in a cross-sectional study. Serum samples were tested for the presence of anti-HEV IgG antibodies by enzyme immunoassay (Wantai anti-HEV IgG, Beijing, China) and for the presence of HEV RNA using real time reverse transcriptase-polymerase chain reaction with primers targeting the HEV ORF2 and ORF3. Clinical and laboratory tests as well as abdominal ultrasound were performed at the same day of blood collection. Results Anti-HEV IgG was positive in 18.8% (15/80) of patients with SM. None of the samples tested positive for anti-HEV IgM or HEV-RNA. Patients with anti-HEV IgG positive presented higher levels of alanine aminotranferase (p = 0.048) and gama-glutamil transferase (p = 0.022) when compared to patients without anti-HEV IgG antibodies. Conclusion This study demonstrates that the seroprevalence of HEV is high in patients with Schistosoma mansoni in Northeastern of Brazil. Past HEV infection is associated with higher frequency of liver enzymes abnormalities. HEV infection and its role on the severity of liver disease should be further investigated among patients with Schistosoma mansoni.

A Korean patient with Guillain-Barré syndrome following acute hepatitis E whose cholestasis resolved with steroid therapy

Autochthonous hepatitis E virus (HEV) is an emerging pathogen in developed countries, and several cases of acute HEV infection have been reported in South Korea. However, there have been no reports on HEV-associated Guillain-Barré syndrome (GBS) in Korea. We recently experienced the case of a 58-year-old Korean male with acute HEV infection after ingesting raw deer meat. Persistent cholestasis was resolved by the administration of prednisolone. At 2.5 months after the clinical presentation of HEV infection, the patient developed weakness of the lower limbs, and was diagnosed with GBS associated with acute hepatitis E. To our knowledge, this is the second report on supportive steroid therapy for persistent cholestasis due to hepatitis E, and the first report of GBS in a Korean patient with acute HEV infection.

Hepatitis E virus infection in Brazil: results of laboratory-based surveillance from 1998 to 2013

Data on hepatitis E virus (HEV) in Brazil are limited. We analyzed 15 years of HEV surveillance data in a major clinical laboratory in São Paulo, Brazil.

Molecular detection of hepatitis E virus in feces and slurry from swine farms, Rio Grande do Sul, Southern Brazil / Detecção molecular do vírus da hepatite E em fezes e chorume de propriedades suinícolas, Rio Grande do Sul, Sul do Brasil

Hepatitis E virus (HEV) is highly disseminated among swine herds worldwide. HEV is also a threat to public health, since particularly genotypes 3 and 4 may cause acute hepatitis in human beings. No previous studies were done on the occurrence of HEV in environmental samples in Rio Grande do Sul, Brazil. In the present study, reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to detect the presence of HEV in swine feces and in effluents from slurry lagoons in farms located in the municipality of Teutônia, inside the area of swine husbandry in the state. Pooled fecal samples from the floor of pig barns from 9 wean-to-finish farms and liquid manure samples were collected from the slurry lagoons from 8 of these farms. From the pooled fecal samples, 8/9 were positive for the HEV ORF1 gene by RT-PCR; all the slurry lagoon samples were positive for HEV RNA (100%). The identity of the HEV ORF1 amplicons was confirmed by sequencing belonging to HEV genotype 3, which was previously shown to be circulating in South America.

O vírus da hepatite E (HEV) é altamente disseminado entre rebanhos suínos no mundo todo. O HEV é também uma ameaça à saúde pública, já que os genótipos 3 e 4 podem causar hepatite aguda em seres humanos. Não há estudos anteriores sobre a ocorrência de HEV em amostras ambientais no Rio Grande do Sul. No presente estudo, empregou-se transcrição reversa e reação em cadeia da polimerase (RT-PCR) para detectar a presença de HEV em fezes de suínos e efluentes de lagoas de chorume em fazendas localizadas no município de Teutônia, representativo da região de maior produção de suínos no estado. Pools de amostras fecais foram coletadas a partir do chão de galpões de suínos provenientes de 9 propriedades de terminação; outra amostra de esterco líquido foi coletada das lagoas de chorume de 8 dessas fazendas. A partir das amostras fecais reunidas, 8/9 foram positivas para o gene ORF1 de HEV por PCR convencional; todas as amostras de lagoas de chorume foram positivas para RNA de HEV (100%). A identificação dos produtos de amplificação de HEV ORF1 foi confirmada por sequenciamento pertencente ao HEV genótipo 3, o qual foi previamente detectado na América do Sul.

A Survey on the Status of Hepatitis E Virus Infection Among Slaughterhouse Workers in South Korea / 예방의학회지

OBJECTIVES: The seroprevalence of hepatitis E virus (HEV) among high-risk groups overseas is high, but studies in these groups are rare in South Korea. We conducted the present study from April to November 2012 to obtain data on the seroprevalence and associated risk factors for HEV among slaughterhouse workers in South Korea. METHODS: Slaughterhouse workers from 80 workplaces nationwide were surveyed in South Korea in 2012. The subjects comprised 1848 cases: 1434 slaughter workers and 414 residual products handlers. By visiting 80 slaughterhouses, which were mixed with 75 of which also performed residual products handling, we conducted a questionnaire survey for risk factors and obtained blood samples in order to determine the seropositivity and seroprevalence of HEV. Anti-HEV IgG and IgM were measured using HEV IgG and IgM enzyme-linked immunospecific assay kits and HEV antigen was measured by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The seropositivity of anti-HEV IgG was 33.5% (slaughter workers 32.8% and residual products handlers 36.2%), and among the seropositive individuals the seroprevalence of anti-HEV IgM was 0.5% (slaughter workers 0.5%, residual products handlers 0.7%). The response rate of HEV-antigen as measured by RT-PCR was 0.2%. Risk factors significantly related to anti-HEV IgG seropositivity were age, sex , and working duration (slaughter workers only). CONCLUSIONS: There were significant risk factors (sex, age, and working duration) for HEV identified in our study. All three positive cases for HEV-antigen by RT-PCR were related to pig slaughter but without statistical significance. To prevent HEV, an educational program and working guidelines may be needed for high risk groups.

Infección por el virus de la hepatitis E en pacientes con diagnóstico clínico de hepatitis viral en Colombia / Hepatitis E virus infection in patients with clinical diagnosis of viral hepatitis in Colombia

Introducción. El virus de la hepatitis E (HEV), agente etiológico de casos esporádicos y epidemias de hepatitis, es un virus emergente de importancia global. En Colombia se desconoce la epidemiología de la infección causada por este virus. Objetivo. Determinar la seropositividad para el virus de la hepatitis E en muestras de suero de pacientes con diagnóstico clínico de hepatitis viral en Colombia. Materiales y métodos. Se estudiaron muestras de pacientes remitidas al Instituto Nacional de Salud en el periodo 2005-2010 provenientes de 15 departamentos de Colombia (grupo 1) y muestras remitidas al Laboratorio Departamental de Salud Pública de Antioquia en el periodo 2008-2009 (grupo 2). Las muestras de suero se analizaron por inmunoensayo con estuches comerciales. Resultados. La frecuencia de seropositividad para el virus de la hepatitis E en las 344 muestras analizadas fue de 8,7 % (30/344); de estas, 1,74 % (6/344) presentó IgM anti-HEV y 7,5 % (26/344), IgG anti-HEV. Se observó una diferencia en el resultado positivo entre el grupo 1 (6,3 %) y el grupo 2 (15,3 %). Los casos provenían de nueve departamentos del país. Conclusiones. Este es el primer estudio de infección por el virus de la hepatitis E en muestras de pacientes con diagnóstico de hepatitis en Colombia. La seropositividad descrita en esta población de pacientes es similar a la descrita en otros países de América Latina, como Brasil, Perú y Uruguay. Teniendo en cuenta estos resultados, se debe considerar la inclusión de los marcadores de la infección por el virus de la hepatitis E en el diagnóstico diferencial de la hepatitis viral en Colombia.

Introduction: Hepatitis E virus (HEV) is an emergent virus of global importance; it is the etiological agent of sporadic cases and outbreaks of hepatitis. The epidemiology of this infection in Colombia is unknown. Objective: To determine the seropositivity for hepatitis E virus in Colombia in cases with clinical diagnosis of viral hepatitis. Materials and methods: Serum samples from patients that were sent to the Instituto Nacional de Salud during the period 2005-2010 (group 1) and samples sent to the Laboratorio Departamental de Salud Pública de Antioquia during the 2008-2009 period were included in this study (group 2). Serum samples were analyzed by immunoassay with commercial kits. Results: From the 344 analyzed samples, 8.7% were positive for anti-HEV; the frequency of anti-HEV IgM was 1.74% (6/344) and the frequency of anti-HEV IgG was 7.5% (26/344). A difference in frequency of anti-HEV between group 1 (6.3%) and group 2 (1.3%) was observed. The cases were identified in nine departments of Colombia. Conclusions: This is the first study of hepatitis E virus infection in patients with diagnosis of hepatitis in Colombia. The frequency of anti-HEV described in this population of patients in Colombia is similar to that described in other Latin American countries like Brazil, Perú and Uruguay. Considering the results of this study, it could be necessary to include hepatitis E virus infection serological markers in the differential diagnosis of viral hepatitis in Colombia.

Hepatitis E: un virus infrecuente o un agente infrecuentemente pensado? / [Hepatitis E: an infrequent virus or an unfrecuently investigated agent?].

Two cases of hepatitis E that were assisted in our Unit of Hepatology and Liver Transplantation are described in this article. The first patient had prior hepatic involvement and therefore a more severe course, whereas the second case, with normal liver function, only developed a self-limited acute hepatitis.

Development of a novel enzyme-linked immunosorbent assay to detect anti-IgG against swine hepatitis E virus

Swine hepatitis E virus (HEV) is widespread throughout pigs in both developing and industrialized countries. This virus is an important zoonotic agent and a public concern worldwide. Infected pigs are asymptomatic, so diagnosing swine HEV relies on detection of the virus or antibodies against the virus. However, several obstacles need to be overcome for effective and practical serological diagnosis. In this study, we developed an enzyme-linked immunosorbent assay (ELISA) that used a purified recombinant capsid protein of swine HEV. The potential clinical use of this assay was evaluated by comparing it with a commercial kit (Genelabs Technologies, Diagnostics, Singapore). Results of the ELISA were highly correlated with those of the commercial kit with a sensitivity of 97% and specificity of 95%. ROC (receiving operator characteristic) analysis of the ELISA data produced a value of 0.987 (95% CI, 0.977~0.998, p < 0.01). The cut-off value for the ELISA was also determined using negative pig sera. In summary, the HEV-specific ELISA developed in the present study appears to be both practical and economical.

Genotype-4 hepatitis E in a human after ingesting roe deer meat in South Korea

The recent increase in the number of cases of indigenous hepatitis E virus (HEV) infection highlights the importance of identifying the transmission routes for the prevention of such infections. Presented herein is the first case of acute HEV infection after ingesting wild roe deer meat in South Korea. A 43-year-old male presented with abdominal discomfort and jaundice. He had not recently traveled abroad, but had eaten raw roe-deer meat 6-8 weeks before the presentation. On the 7th day of hospitalization the patient was diagnosed with acute viral hepatitis E. Phylogenetic analysis of his serum revealed genotype-4 HEV. This case supports the possibility of zoonotic transmission of HEV because the patient appears to have been infected with genotype-4 HEV after ingesting raw deer meat.

Prevalence of hepatitis E virus antibodies in individuals exposed to swine in Mato Grosso, Brazil

This study was conducted to estimate the prevalence of hepatitis E antibodies (anti-HEV) among individuals exposed to swine in the rural areas of the state of Mato Grosso (MT) in Brazil. The study included 310 participants who had an average age of 39 years. Fifty-one per cent of the participants were female and 26 (8.4%) were anti-HEV-positive. Concomitantly, we studied 101 blood donors from the urban area of the state capital who had never lived in a rural area or handled swine. Four per cent (4%) of these individuals were anti-HEV-positive (p = 0.206). When we compared the anti-HEV-positive participants who had been exposed to swine with the anti-HEV-negative participants, we noticed associations between the presence of anti-HEV and increased age, a history of blood transfusions and contact with other farm animals. However, after a multivariate analysis was performed, this association was not confirmed. Finally, the ratio of anti-HEV-positive individuals who had been exposed to swine in rural MT was similar to that found in previous studies in Brazil. This prevalence did not characterise this type of exposure as a risk factor for HEV infection in this region.

Prolonged jaundice attributed to super infection of hepatitis E virus in a case of resolving leptospirosis.

India is endemic for both Leptospira and hepatitis E virus (HEV). The clinical presentations of these diseases have overlapping features. We report a case of superinfection of HEV in a patient with resolving leptospirosis with underlying Hodgkin lymphoma. The diagnosis of HEV in our case was established by HEV-RNA PCR as our patient was immunosuppressed. The present study highlights the need for molecular diagnosis in the case of HEV infection with strong clinical suspicion and negative serological results.