Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 14 de 14
Filtrar
1.
Appl Microbiol Biotechnol ; 105(12): 4957-4973, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34129082

RESUMO

To generate a hepatitis E virus (HEV) genotype 3 (HEV-3)-specific monoclonal antibody (mAb), the Escherichia coli-expressed carboxy-terminal part of its capsid protein was used to immunise BALB/c mice. The immunisation resulted in the induction of HEV-specific antibodies of high titre. The mAb G117-AA4 of IgG1 isotype was obtained showing a strong reactivity with the homologous E. coli, but also yeast-expressed capsid protein of HEV-3. The mAb strongly cross-reacted with ratHEV capsid protein derivatives produced in both expression systems and weaker with an E. coli-expressed batHEV capsid protein fragment. In addition, the mAb reacted with capsid protein derivatives of genotypes HEV-2 and HEV-4 and common vole hepatitis E virus (cvHEV), produced by the cell-free synthesis in Chinese hamster ovary (CHO) and Spodoptera frugiperda (Sf21) cell lysates. Western blot and line blot reactivity of the mAb with capsid protein derivatives of HEV-1 to HEV-4, cvHEV, ratHEV and batHEV suggested a linear epitope. Use of truncated derivatives of ratHEV capsid protein in ELISA, Western blot, and a Pepscan analysis allowed to map the epitope within a partially surface-exposed region with the amino acid sequence LYTSV. The mAb was also shown to bind to human patient-derived HEV-3 from infected cell culture and to hare HEV-3 and camel HEV-7 capsid proteins from transfected cells by immunofluorescence assay. The novel mAb may serve as a useful tool for further investigations on the pathogenesis of HEV infections and might be used for diagnostic purposes. KEY POINTS: • The antibody showed cross-reactivity with capsid proteins of different hepeviruses. • The linear epitope of the antibody was mapped in a partially surface-exposed region. • The antibody detected native HEV-3 antigen in infected mammalian cells.


Assuntos
Vírus da Hepatite E , Animais , Anticorpos Monoclonais , Células CHO , Capsídeo , Proteínas do Capsídeo , Cricetinae , Cricetulus , Escherichia coli , Humanos , Camundongos , Camundongos Endogâmicos BALB C
2.
Med Microbiol Immunol ; 203(4): 273-82, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24744181

RESUMO

An increase in acute autochthonous hepatitis E virus (HEV) infections has been recorded in Germany. These are suspected to be zoonotically transmitted from wild boar, deer and domestic pig. The latter may represent a major reservoir for HEV. In this study, 537 sera from humans living in Westphalia and Lower Saxony, representing areas of high pig density in Germany, were tested for the presence of HEV-specific antibodies. Among them were 302 individuals with occupational, direct contact to pigs and 235 individuals without direct contact to pigs. Two commercial tests and one in-house assay were applied for the detection of HEV-specific immunoglobulin G (IgG) antibodies. Sera were also tested in an assay that detects all classes of HEV-specific antibodies. Depending on the test used, the seroprevalence ranged from 4.1 to 27.9 %. Exposition to pigs was found to be associated with a significantly higher seroprevalence in subjects with contact to pigs (13.2-32.8 %) compared with that in non-exposed humans (7.7-21.7 %). In particular, individuals younger than 40 years with occupational exposure exhibited a markedly higher HEV seroprevalence compared with non-exposed individuals of that age group. In general, HEV seroprevalence increased with age resulting in a similar prevalence level in the age group of ≥ 50 years for exposed and non-exposed individuals. Analysis of all sera by a commercial anti-HEV IgM ELISA revealed 35 positive and 25 borderline samples. However, only one positive serum could be confirmed by an IgM line assay. Selected samples from IgM and/or IgG as well as total HEV antibody-positive individuals were also tested for the presence of HEV RNA. In one of the 78 samples, the only IgM ELISA positive and IgM line assay confirmed sample, RNA of HEV genotype 3 was detected. This sequence has high similarity to HEV sequences obtained from wild boars and domestic pigs from Germany and The Netherlands. This study demonstrates that in addition to the consumption of raw or undercooked meat, direct contact to pigs has to be considered as an additional risk factor for HEV infection.


Assuntos
Reservatórios de Doenças , Anticorpos Anti-Hepatite/sangue , Vírus da Hepatite E/imunologia , Hepatite E/epidemiologia , Exposição Ocupacional , Sus scrofa , Zoonoses/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Feminino , Alemanha/epidemiologia , Humanos , Imunoglobulina G/sangue , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Estudos Soroepidemiológicos , Adulto Jovem
3.
Circ Res ; 110(8): e50-63, 2012 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-22427340

RESUMO

RATIONALE: Innate and adaptive immune responses alter numerous homeostatic processes that are controlled by nuclear hormone receptors. NR4A1 is a nuclear receptor that is induced in vascular pathologies, where it mediates protection. OBJECTIVE: The underlying mechanisms that regulate the activity of NR4A1 during vascular injury are not clear. We therefore searched for modulators of NR4A1 function that are present during vascular inflammation. METHODS AND RESULTS: We report that the protein encoded by interferon stimulated gene 12 (ISG12), is a novel interaction partner of NR4A1 that inhibits the transcriptional activities of NR4A1 by mediating its Crm1-dependent nuclear export. Using 2 models of vascular injury, we show that ISG12-deficient mice are protected from neointima formation. This effect is dependent on the presence of NR4A1, as mice deficient for both ISG12 and NR4A1 exhibit neointima formation similar to wild-type mice. CONCLUSIONS: These findings identify a previously unrecognized feedback loop activated by interferons that inhibits the vasculoprotective functions of NR4A nuclear receptors, providing a potential new therapeutic target for interferon-driven pathologies.


Assuntos
Lesões das Artérias Carótidas/prevenção & controle , Artéria Femoral/metabolismo , Inflamação/prevenção & controle , Proteínas de Membrana/metabolismo , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Proteínas/metabolismo , Lesões do Sistema Vascular/prevenção & controle , Transporte Ativo do Núcleo Celular , Animais , Lesões das Artérias Carótidas/genética , Lesões das Artérias Carótidas/imunologia , Lesões das Artérias Carótidas/metabolismo , Lesões das Artérias Carótidas/patologia , Células Cultivadas , Modelos Animais de Doenças , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Retroalimentação Fisiológica , Artéria Femoral/lesões , Artéria Femoral/patologia , Regulação da Expressão Gênica , Inflamação/genética , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/patologia , Interferons/metabolismo , Carioferinas/metabolismo , Proteínas de Membrana/genética , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Músculo Liso Vascular/lesões , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Domínios e Motivos de Interação entre Proteínas , Proteínas/genética , Interferência de RNA , Receptores Citoplasmáticos e Nucleares/metabolismo , Fatores de Tempo , Transcrição Gênica , Transfecção , Lesões do Sistema Vascular/genética , Lesões do Sistema Vascular/imunologia , Lesões do Sistema Vascular/metabolismo , Lesões do Sistema Vascular/patologia , Proteína Exportina 1
4.
Vet Res ; 45: 121, 2014 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-25421429

RESUMO

Hepatitis E virus (HEV) is the causative agent of acute hepatitis E in humans in developing countries, but sporadic and autochthonous cases do also occur in industrialised countries. In Europe, food-borne zoonotic transmission of genotype 3 (gt3) has been associated with domestic pig and wild boar. However, little is known about the course of HEV infection in European wild boar and their role in HEV transmission to domestic pigs. To investigate the transmissibility and pathogenesis of wild boar-derived HEVgt3, we inoculated four wild boar and four miniature pigs intravenously. Using quantitative real-time RT-PCR viral RNA was detected in serum, faeces and in liver, spleen and lymph nodes. The antibody response evolved after fourteen days post inoculation. Histopathological findings included mild to moderate lymphoplasmacytic hepatitis which was more prominent in wild boar than in miniature pigs. By immunohistochemical methods, viral antigens were detected mainly in Kupffer cells and liver sinusoidal endothelial cells, partially associated with hepatic lesions, but also in spleen and lymph nodes. While clinical symptoms were subtle and gross pathology was inconspicuous, increased liver enzyme levels in serum indicated hepatocellular injury. As the faecal-oral route is supposed to be the most likely transmission route, we included four contact animals to prove horizontal transmission. Interestingly, HEVgt3-infection was also detected in wild boar and miniature pigs kept in contact to intravenously inoculated wild boar. Given the high virus loads and long duration of viral shedding, wild boar has to be considered as an important HEV reservoir and transmission host in Europe.


Assuntos
Vírus da Hepatite E/fisiologia , Hepatite E/veterinária , Doenças dos Suínos/transmissão , Animais , Anticorpos Antivirais/sangue , Formação de Anticorpos , Genótipo , Hepatite E/transmissão , Hepatite E/virologia , Vírus da Hepatite E/genética , RNA Viral/sangue , RNA Viral/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Sus scrofa , Suínos , Doenças dos Suínos/virologia , Porco Miniatura
5.
Med Microbiol Immunol ; 201(2): 189-200, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22179131

RESUMO

Hepatitis E virus (HEV) is the causative agent of an acute self-limiting hepatitis in humans. In industrialized countries, autochthonous cases are linked to zoonotic transmission from domestic pigs, wild boar and red deer. The main route of human infection presumably is consumption of contaminated meat. Farmers, slaughterers and veterinarians are expected to be risk groups as they work close to potentially infected animals. In this study, we tested four Escherichia coli-expressed segments of the capsid protein (CP) of a German wild boar-derived HEV genotype 3 strain for their diagnostic value in an indirect immunoglobulin G (IgG) ELISA. In an initial validation experiment, a carboxy-terminal CP segment spanning amino acid (aa) residues 326-608 outperformed the other segments harbouring aa residues 112-608, 326-660 and 112-335. Based on this segment, an indirect ELISA for detection of anti-HEV IgG antibodies in human sera was established and validated using a commercial line immunoassay as reference assay. A total of 563 sera from forestry workers of all forestry offices of Brandenburg, eastern Germany and 301 sera of blood donors from eastern Germany were surveyed using these assays. The commercial test revealed seroprevalence rates of 11% for blood donors and 18% for forestry workers. These rates are in line with data obtained by the in-house test (12 and 21%). Hence, the in-house test performed strikingly similar to the commercial test (sensitivity 0.9318, specificity 0.9542). An initial screening of forestry worker and blood donor sera with a corresponding CP segment of the recently discovered Norway rat-associated HEV revealed several strong positive sera exclusively in the forestry worker panel. Future investigations have to prove the performance of this novel IgG ELISA in large-scale seroepidemiological studies. In addition, the observed elevated seroprevalence in a forestry worker group has to be confirmed by studies on groups of forestry workers from other regions. The epidemiological role of ratHEV in human disease should be assessed in a large-scale study of risk and non-risk groups.


Assuntos
Agricultura , Agricultura Florestal , Anticorpos Anti-Hepatite/sangue , Vírus da Hepatite E/imunologia , Hepatite E/epidemiologia , Exposição Ocupacional , Animais , Técnicas de Laboratório Clínico/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Genótipo , Alemanha/epidemiologia , Vírus da Hepatite E/genética , Humanos , Imunoglobulina G/sangue , Masculino , Ratos/virologia , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Sus scrofa/virologia
6.
Genes (Basel) ; 12(12)2021 12 08.
Artigo em Inglês | MEDLINE | ID: mdl-34946907

RESUMO

In recent years, optical genome mapping (OGM) has developed into a highly promising method of detecting large-scale structural variants in human genomes. It is capable of detecting structural variants considered difficult to detect by other current methods. Hence, it promises to be feasible as a first-line diagnostic tool, permitting insight into a new realm of previously unknown variants. However, due to its novelty, little experience with OGM is available to infer best practices for its application or to clarify which features cannot be detected. In this study, we used the Saphyr system (Bionano Genomics, San Diego, CA, USA), to explore its capabilities in human genetic diagnostics. To this end, we tested 14 DNA samples to confirm a total of 14 different structural or numerical chromosomal variants originally detected by other means, namely, deletions, duplications, inversions, trisomies, and a translocation. Overall, 12 variants could be confirmed; one deletion and one inversion could not. The prerequisites for detection of similar variants were explored by reviewing the OGM data of 54 samples analyzed in our laboratory. Limitations, some owing to the novelty of the method and some inherent to it, were described. Finally, we tested the successful application of OGM in routine diagnostics and described some of the challenges that merit consideration when utilizing OGM as a diagnostic tool.


Assuntos
Aberrações Cromossômicas , Transtornos Cromossômicos/diagnóstico , Mapeamento Cromossômico/métodos , Mapeamento Cromossômico/normas , Variações do Número de Cópias de DNA , Genoma Humano , Cariotipagem/métodos , Transtornos Cromossômicos/genética , Feminino , Humanos , Masculino
7.
Genes (Basel) ; 12(11)2021 11 21.
Artigo em Inglês | MEDLINE | ID: mdl-34828442

RESUMO

Marfan syndrome (MFS) is a hereditary connective tissue disease caused by heterozygous mutations in the fibrillin-1 gene (FBN1) located on chromosome 15q21.1. A complex chromosomal rearrangement leading to MFS has only been reported in one case so far. We report on a mother and daughter with marfanoid habitus and no pathogenic variant in the FBN1 gene after next generation sequencing (NGS) analysis, both showing a cytogenetically reciprocal balanced translocation between chromosomes 2 and 15. By means of fluorescence in situ hybridization of Bacterial artificial chromosome (BAC) clones from the breakpoint area on chromosome 15 the breakpoint was narrowed down to a region of approximately 110 kb in FBN1. With the help of optical genome mapping (OGM), the translocation breakpoints were further refined on chromosomes 2 and 15. Sequencing of the regions affected by the translocation identified the breakpoint of chromosome 2 as well as the breakpoint of chromosome 15 in the FBN1 gene leading to its disruption. To our knowledge, this is the first report of patients with typical clinical features of MFS showing a cytogenetically reciprocal translocation involving the FBN1 gene. Our case highlights the importance of structural genome variants as an underlying cause of monogenic diseases and the useful clinical application of OGM in the elucidation of structural variants.


Assuntos
Fibrilina-1/genética , Síndrome de Marfan/genética , Translocação Genética , Adolescente , Adulto , Pontos de Quebra do Cromossomo , Humanos , Masculino , Síndrome de Marfan/patologia , Linhagem
8.
Viruses ; 11(1)2018 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-30577433

RESUMO

Hepatitis E virus (HEV) is the causative agent of acute hepatitis E in humans in developing countries, but autochthonous cases of zoonotic genotype 3 (HEV-3) infection also occur in industrialized countries. In contrast to swine, rats, and rabbits, natural HEV infections in mice have not yet been demonstrated. The pig represents a well-established large animal model for HEV-3 infection, but a suitable small animal model mimicking natural HEV-3 infection is currently missing. Therefore, we experimentally inoculated C57BL/6 mice (wild-type, IFNAR-/-, CD4-/-, CD8-/-) and BALB/c nude (nu/nu) mice, Wistar rats, and European rabbits with a wild boar-derived HEV-3 strain and monitored virus replication and shedding, as well as humoral immune responses. HEV RNA and anti-HEV antibodies were detected in one and two out of eight of the rats and all rabbits inoculated, respectively, but not in any of the mouse strains tested. Remarkably, immunosuppressive dexamethasone treatment of rats did not enhance their susceptibility to HEV infection. In rabbits, immunization with recombinant HEV-3 and ratHEV capsid proteins induced protection against HEV-3 challenge. In conclusion, the rabbit model for HEV-3 infection may serve as a suitable alternative to the non-human primate and swine models, and as an appropriate basis for vaccine evaluation studies.


Assuntos
Modelos Animais de Doenças , Hepatite E/imunologia , Imunidade Humoral , Replicação Viral , Eliminação de Partículas Virais , Animais , Dexametasona/administração & dosagem , Fezes/virologia , Feminino , Genótipo , Anticorpos Anti-Hepatite/sangue , Hepatite E/prevenção & controle , Vírus da Hepatite E/genética , Vírus da Hepatite E/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , RNA Viral , Coelhos , Ratos , Ratos Wistar , Vacinas Sintéticas/imunologia , Vacinas contra Hepatite Viral/imunologia
9.
Vet Microbiol ; 208: 58-68, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28888650

RESUMO

Rat hepatitis E virus (HEV) is genetically only distantly related to hepeviruses found in other mammalian reservoirs and in humans. It was initially detected in Norway rats (Rattus norvegicus) from Germany, and subsequently in rats from Vietnam, the USA, Indonesia, China, Denmark and France. Here, we report on a molecular survey of Norway rats and Black rats (Rattus rattus) from 12 European countries for ratHEV and human pathogenic hepeviruses. RatHEV-specific real-time and conventional RT-PCR investigations revealed the presence of ratHEV in 63 of 508 (12.4%) rats at the majority of sites in 11 of 12 countries. In contrast, a real-time RT-PCR specific for human pathogenic HEV genotypes 1-4 and a nested broad-spectrum (NBS) RT-PCR with subsequent sequence determination did not detect any infections with these genotypes. Only in a single Norway rat from Belgium a rabbit HEV-like genotype 3 sequence was detected. Phylogenetic analysis indicated a clustering of all other novel Norway and Black rat-derived sequences with ratHEV sequences from Europe, the USA and a Black rat-derived sequence from Indonesia within the proposed ratHEV genotype 1. No difference in infection status was detected related to age, sex, rat species or density of human settlements and zoological gardens. In conclusion, our investigation shows a broad geographical distribution of ratHEV in Norway and Black rats from Europe and its presence in all settlement types investigated.


Assuntos
Vírus da Hepatite E/classificação , Vírus da Hepatite E/isolamento & purificação , Hepatite E/veterinária , Distribuição Animal , Animais , Animais Selvagens , Europa (Continente)/epidemiologia , Feminino , Hepatite E/epidemiologia , Hepatite E/virologia , Vírus da Hepatite E/genética , Humanos , Masculino , Filogenia , Densidade Demográfica , Ratos , Especificidade da Espécie
10.
J Wildl Dis ; 52(1): 106-13, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26528571

RESUMO

Hepatitis E virus (HEV) is a human pathogen that is primarily transmitted by the fecal-oral route and causes a usually self-limiting acute viral hepatitis. The virus is endemic in developing countries of Africa, Asia, and Latin America and is responsible for sporadic cases in industrialized countries. In western Europe, an increasing number of autochthonous cases have been associated with zoonotic transmissions of HEV from domestic and wild animals. In Germany, animal reservoirs for HEV have been mainly assigned to domestic pigs and wild boars. To investigate the potential role of deer as a reservoir of HEV, we surveyed HEV-specific antibodies and RNA in deer samples from geographic regions in Germany. We sampled red deer (Cervus elaphus) and roe deer (Capreolus capreolus) during active surveillance in three forest districts in northern Hesse and southern Lower Saxony during 2011-12 and 2012-13, respectively. Additionally, archived samples of red, roe, and fallow deer (Dama dama), collected in 2000-01 in German national parks, were included in the study. Antibody prevalence ranged from 2-3.3% in red deer to 5.4-6.8% in roe deer. Viral RNA was detected in red deer and fallow deer at prevalences of 2.0-6.6% and 4.3%, respectively. The investigation confirmed the presence of HEV infections in three deer species in Germany. Red, roe, and fallow deer should be further monitored to assess their role as hosts and potential reservoirs of HEV in Germany.


Assuntos
Cervos , Reservatórios de Doenças/veterinária , Vírus da Hepatite E/isolamento & purificação , Hepatite E/veterinária , Animais , Anticorpos Antivirais/sangue , Cervos/classificação , Reservatórios de Doenças/virologia , Feminino , Alemanha/epidemiologia , Hepatite E/epidemiologia , Hepatite E/transmissão , Vírus da Hepatite E/genética , Vírus da Hepatite E/imunologia , Imunoglobulina G/sangue , Fígado/virologia , Masculino , Prevalência , RNA Viral/análise , Reação em Cadeia da Polimerase em Tempo Real/veterinária
11.
Infect Genet Evol ; 27: 212-29, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25050488

RESUMO

The hepatitis E virus (HEV) was first identified in 1990, although hepatitis E-like diseases in humans have been recorded for a long time dating back to the 18th century. The HEV genotypes 1-4 have been subsequently detected in human hepatitis E cases with different geographical distribution and different modes of transmission. Genotypes 3 and 4 have been identified in parallel in pigs, wild boars and other animal species and their zoonotic potential has been confirmed. Until 2010, these genotypes along with avian HEV strains infecting chicken were the only known representatives of the family Hepeviridae. Thereafter, additional HEV-related viruses have been detected in wild boars, distinct HEV-like viruses were identified in rats, rabbit, ferret, mink, fox, bats and moose, and a distantly related agent was described from closely related salmonid fish. This review summarizes the characteristics of the so far known HEV-like viruses, their phylogenetic relationship, host association and proposed involvement in diseases. Based on the reviewed knowledge, a suggestion for a new taxonomic grouping scheme of the viruses within the family Hepeviridae is presented.


Assuntos
Vírus da Hepatite E/classificação , Vírus da Hepatite E/genética , Hepatite E/virologia , Zoonoses/virologia , Animais , Genoma Viral , Hepatite E/transmissão , Interações Hospedeiro-Patógeno , Humanos , Vertebrados , Zoonoses/transmissão
12.
Vet Microbiol ; 167(3-4): 394-402, 2013 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-24238666

RESUMO

An increasing number of acute autochthonous human hepatitis E virus (HEV)-infections was noticed in Germany and other developed countries, most likely the result of a zoonotic virus transmission from pig, wild boar and deer. Currently there is still a lack of profound data concerning the actual prevalence of HEV-specific antibodies in domestic pig herds in Germany, in particular for regions with high pig density, and its age-dependency. 2273 domestic pig sera were collected in 2011 mainly from Bavaria, North Rhine-Westphalia and Lower Saxony from areas having a high pig density. Initially, 420 randomly selected pig sera were tested in three commercially available and in two in-house HEV-antibody ELISAs. 43.6% (183/420) to 65.5% (275/420) of the sera were demonstrated to be reactive against human pathogenic HEV genotypes 1 and/or 3. The majority of sera reacted only weakly or not at all with the rat HEV antigen with very few sera showing a stronger reactivity to this antigen compared to the genotype 3 antigen. The results of all three HEV-IgG tests, i.e. the PrioCHECK(®) HEV Ab porcine ELISA kit, the ID Screen(®) Hepatitis E Indirect Multi-species ELISA kit and the genotype 3 in-house ELISA were in good accordance. Therefore, the remaining sera were tested using the PrioCHECK(®) HEV Ab porcine ELISA kit. Samples with a borderline result were finally determined by application of the conjugate-modified recomLine HEV IgG assay. A total of 1065 of the 2273 sera (46.9%) were found to be anti-HEV IgG-positive. While 38.4% (306/796) of fatteners (age between 3 and 9 months) exhibited HEV-specific antibodies, 51.4% (759/1477) of sows (age older than 9 months) exhibited anti-HEV antibodies (P<0.001). Fatteners kept in Southern Germany had a significantly higher HEV IgG prevalence compared to fatteners kept in the high pig density federal states North Rhine-Westphalia and Lower Saxony but also in German federal states with a low pig density. In conclusion, the present study clearly demonstrates that a high percentage of domestic pigs in Germany have had contact with HEV. Seroprevalence depends on the pig's age and herd origin with the most significant regional variations for fatteners. The presence of anti-HEV-free herds may indicate that it is feasible to establish and sustain HEV-free pig herds. HEV seroprevalence still depends on the assay used for testing. This demonstrates an urgent need for test validation.


Assuntos
Anticorpos Anti-Hepatite/sangue , Hepatite E/veterinária , Doenças dos Suínos/epidemiologia , Fatores Etários , Animais , Geografia , Alemanha/epidemiologia , Hepatite E/epidemiologia , Vírus da Hepatite E/fisiologia , Imunoglobulina G/sangue , Estudos Soroepidemiológicos , Testes Sorológicos/normas , Testes Sorológicos/veterinária , Sus scrofa/virologia , Suínos
13.
J Virol Methods ; 190(1-2): 11-6, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23523888

RESUMO

Autochthonous hepatitis E virus (HEV) infections by zoonotic transmission of genotype 3 (GT3) have been reported increasingly from industrialized countries. In this paper the development and validation of an IgG ELISA for the detection of HEV-specific antibodies in domestic pigs is described. Comparison of the diagnostic value of Escherichia coli-expressed HEV-GT3 capsid protein (CP) derivatives revealed a carboxy-terminal derivative as most suitable. Validation of the in-house assay using a commercially available IgG ELISA revealed a high diagnostic specificity and sensitivity. The average HEV seroprevalence of domestic pigs from Germany and the federal state Baden-Wuerttemberg determined by the in-house test was 42.7% and 50.3%, respectively. The seroprevalence in different districts of Baden-Wuerttemberg ranged from 34.9% to 60%, but from 0% to 100% between different herds. These data were compared to those achieved by two commercially available ELISA kits and an in-house ratHEV-based ELISA. In conclusion, the CP-based in-house test proved sensitive and specific, indicating that the ORF3-encoded protein might be dispensable for diagnostics. The novel assay also allowed a parallel analysis by a homologous ratHEV-derived antigen. Thus, the novel IgG ELISA represents a useful tool for future standardized seroprevalence studies in domestic pigs from Germany and other regions of Europe.


Assuntos
Anticorpos Antivirais/sangue , Hepatite E/epidemiologia , Doenças dos Suínos/epidemiologia , Medicina Veterinária/métodos , Virologia/métodos , Animais , Antígenos Virais , Ensaio de Imunoadsorção Enzimática/métodos , Alemanha/epidemiologia , Imunoglobulina G/sangue , Proteínas Recombinantes , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Sus scrofa , Suínos , Doenças dos Suínos/virologia
14.
Infect Genet Evol ; 12(5): 947-56, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22554648

RESUMO

Zoonotic hepatitis E virus (HEV) infection in industrialised countries is thought to be caused by transmission from wild boar, domestic pig and deer as reservoir hosts. The detection of HEV-specific antibodies in rats and other rodents has suggested that these animals may represent an additional source for HEV transmission to human. Recently, a novel HEV (ratHEV) was detected in Norway rats from Hamburg, Germany, showing the typical genome organisation but a high nucleotide and amino acid sequence divergence to other mammalian and to avian HEV strains. Here we describe the multiple detection of ratHEV RNA and HEV-specific antibodies in Norway rats from additional cities in north-east and south-west Germany. The complete genome analysis of two novel strains from Berlin and Stuttgart confirmed the association of ratHEV to Norway rats. The present data indicated a continuing existence of this virus in the rat populations from Berlin and Hamburg. The phylogenetic analysis of a short segment of the open reading frame 1 confirmed a geographical clustering of the corresponding sequences. Serological investigations using recombinant ratHEV and genotype 3 capsid protein derivatives demonstrated antigenic differences which might be caused by the high amino acid sequence divergence in the immunodominant region. The high amount of animals showing exclusively ratHEV RNA or anti-ratHEV antibodies suggested a non-persistent infection in the Norway rat. Future studies have to prove the transmission routes of the virus in rat populations and its zoonotic potential. The recombinant ratHEV antigen generated here will allow future seroepidemiological studies to differentiate ratHEV and genotype 3 infections in humans and animals.


Assuntos
Vírus da Hepatite E/isolamento & purificação , Hepatite E/veterinária , Epidemiologia Molecular , Testes Sorológicos , Animais , Animais Selvagens/virologia , Proteínas do Capsídeo/imunologia , Análise por Conglomerados , Feminino , Genoma Viral/genética , Alemanha/epidemiologia , Anticorpos Anti-Hepatite/sangue , Anticorpos Anti-Hepatite/imunologia , Antígenos de Hepatite/genética , Antígenos de Hepatite/imunologia , Hepatite E/epidemiologia , Hepatite E/imunologia , Hepatite E/virologia , Vírus da Hepatite E/classificação , Vírus da Hepatite E/genética , Vírus da Hepatite E/imunologia , Fígado/imunologia , Fígado/virologia , Filogenia , Cavidade Pleural/imunologia , Cavidade Pleural/virologia , Reação em Cadeia da Polimerase , RNA Viral/análise , RNA Viral/sangue , Ratos , Ratos Wistar , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Virais/genética , Proteínas Virais/imunologia , Zoonoses
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA