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1.
Int J Food Microbiol ; 344: 109089, 2021 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-33662900

RESUMO

Contamination of bivalve molluscs with viruses is well recognized as a food safety risk. A microbiological criterion for norovirus (NoV) and hepatitis A virus (HAV) in shellfish, however, does not exist in the European Union currently. The aim of this study was to evaluate the contamination levels of these viruses for fluctuation over a long period (2013-2017) in oyster (n = 266) and mussel samples (n = 490) using a method based on ISO/TS 15216-1: 2013. Samples were taken at different points in the food chain, either directly post-harvest, at Dutch dispatch centers or in retail stores, from September until March of each year. Altogether, 53.1% of the mussel and 31.6% of the oyster samples tested positive for NoV RNA. Simultaneous presence of NoV GI and GII RNA was observed in 31.6% of mussel and 10.2% of oyster samples. Contamination levels in NoV positive mussel samples collected post-harvest from B-areas were significantly higher than in those collected post-harvest from A-areas, or at dispatch centers or retail stores. Levels in oysters from dispatch were significantly lower than those collected in retail stores. Ready for sale mussels and oysters contained 2.04 and 1.76 mean log10 transformed NoV genome copies/gram (gc/g), respectively. GII levels were at a constant level in ready for sale mussels throughout all sampling periods in the study. This seemed to be true for oysters as well. HAV RNA was detected in only one of the tested mussel samples (n = 392) (typed HAV 1A) and in none of the tested oyster samples (n = 228). Critical evaluation of NoV and HAV levels in shellfish can be of help for risk assessment and risk management actions.


Assuntos
Infecções por Caliciviridae/epidemiologia , Vírus da Hepatite A/isolamento & purificação , Hepatite A/epidemiologia , Norovirus/isolamento & purificação , Ostreidae/virologia , Animais , Infecções por Caliciviridae/veterinária , Cadeia Alimentar , Contaminação de Alimentos/análise , Inocuidade dos Alimentos , Hepatite A/veterinária , Vírus da Hepatite A/genética , Humanos , Países Baixos/epidemiologia , Norovirus/genética , Frutos do Mar/virologia
2.
Viruses ; 13(2)2021 01 29.
Artigo em Inglês | MEDLINE | ID: mdl-33573054

RESUMO

Recently, we experienced an outbreak of acute hepatitis A virus (HAV) infection between 2018 and 2020. Herein, we describe this male-dominant HAV infection outbreak observed among non-human immunodeficiency virus (HIV)-infected persons in the northern part of Tokyo, Japan. Clinical information was collected from patient interviews and from medical record descriptions. In the present study, 21 patients were retrospectively analyzed. A total of 90.4 and 33.3% of patients were males, and men who have sex with men (MSM), respectively. The total bilirubin levels and platelet counts tended to be lower in the MSM group than in the non-MSM group. C-reactive protein (CRP) levels tended to be higher in acute liver failure (ALF) patients than in non-ALF patients. Prolonged cholestasis was observed in one patient (4.8%). We also found that 18 HAV isolates belonged to HAV subgenotype IA/subgroup 13 (S13), which clustered with the HAV isolate (KX151459) that was derived from an outbreak of HAV infection among MSM in Taiwan in 2015. Our results suggest that the application of antivirals against HAV, as well as HAV vaccines, would be useful for the treatment and prevention of severe HAV infection.


Assuntos
Surtos de Doenças , Hepatite A/epidemiologia , Adulto , Feminino , Genótipo , Hepatite A/virologia , Vírus da Hepatite A/classificação , Vírus da Hepatite A/genética , Vírus da Hepatite A/isolamento & purificação , Homossexualidade Masculina , Humanos , Tempo de Internação , Falência Hepática Aguda/epidemiologia , Falência Hepática Aguda/virologia , Masculino , Pessoa de Meia-Idade , Filogenia , Estudos Retrospectivos , Fatores de Risco , Minorias Sexuais e de Gênero , Tóquio/epidemiologia
3.
Int J Food Microbiol ; 337: 108931, 2021 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-33188986

RESUMO

Among the enteric viruses implicated in foodborne outbreaks, the human norovirus and hepatitis viruses A and E (HAV and HEV) represent a serious public health concern. International standard ISO 15216 proposes methods for detecting HAV and norovirus (genogroups I and II) RNA from soft fruit, leaf, stem and bulb vegetables, bottled water or food surfaces. These methods had not previously been validated for detecting the targeted viruses in other foodstuffs such as multicomponent foods, nor for detecting other viruses in foodstuffs. The aim of this study was to characterise a method derived from the vegetable method described in ISO 15216 to detect HAV, HEV and norovirus in artificially-contaminated multicomponent foodstuffs according to the recent international standard ISO 16140-4. Results showed that the mean recovery rates for all settings did not differ according to the operator. The mean extraction yields ranged from 0.35% to 40.44% for HAV, 5.19% to 100% for HEV, 0.10% to 40.61% for norovirus GI and 0.88% to 69.16% for norovirus GII. The LOD95 was 102 genome copies/g for HAV, HEV and norovirus GII and 103 genome copies/g for norovirus GI. The LOQ was 2.90 × 104, 1.40 × 103, 1.60 × 104 and 1.30 × 104 genome copies/g for HAV, HEV, norovirus GI and norovirus GII respectively. The MNV-1 process control was detected in 120 out of 128 RNA extracts analysed and was recovered with an efficiency of between 3.83% and 50.22%. The mean inhibition rates of quantitative real-time RT-PCR reaction ranged from 3.25% to 28.70% and varied significantly with the type of food matrix. The described method could be used to detect viruses in composite food products for routine diagnosis needs.


Assuntos
Microbiologia de Alimentos/métodos , Vírus da Hepatite A/genética , Vírus da Hepatite E/genética , Norovirus/genética , Surtos de Doenças/prevenção & controle , Água Potável/virologia , Frutas/virologia , Vírus da Hepatite A/fisiologia , Limite de Detecção , RNA Viral/análise , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Verduras/virologia
4.
Viruses ; 12(12)2020 12 19.
Artigo em Inglês | MEDLINE | ID: mdl-33352751

RESUMO

Hepatitis A virus (HAV) is a waterborne pathogen of public health importance. In South Africa (SA), unique HAV subgenotype IB strains have been detected in surface and wastewater samples, as well as on fresh produce at the point of retail. However, due to the use of molecular-based assays, the infectivity of the detected strains was unknown. Considering the potential shift of HAV endemicity from high to intermediate, which could increase the risk of severe symptomatic disease, this study investigated the identity of HAV strains detected before and after viability treatment of selected wastewater discharge samples. For one year, 118 samples consisting of sewage, treated wastewater discharge and downstream dam water were collected from five wastewater treatment plants (WWTP 1, 2, 3, 4 and 5). Unique HAV IB strains were detected in samples from all five WWTPs, with 11 of these strains carrying amino acid mutations at the immunodominant and neutralisation epitopes. A quasispecies dynamic of HAV has also been detected in sewage samples. The subsequent application of viability PCR revealed that potentially infectious HAV strains were discharged from WWTP 1, 2, 4 and 5 into the dam. Therefore, there is a potential risk of HAV exposure to communities using water sources downstream the WWTPs.


Assuntos
Vírus da Hepatite A/isolamento & purificação , Esgotos/virologia , Águas Residuárias/virologia , Purificação da Água , Vírus da Hepatite A/genética , Vírus da Hepatite A/imunologia , Filogenia
5.
Viruses ; 13(1)2020 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-33374859

RESUMO

The genetic diversity of Hepatitis A Virus (HAV) circulating in the Campania Region in years 2015-2018 was investigated through the monitoring of sentinel bivalve shellfish and water matrices. Overall, 463 water samples (71 sewage samples, 353 coastal discharge waters, and 39 seawaters samples), and 746 bivalve shellfish samples were analyzed. Positivity for HAV was detected in 20/71 sewage samples, 14/353 coastal discharge waters, 5/39 seawaters, and 102/746 bivalve shellfish. Sixty-one of the positive samples were successfully sequenced and were characterized as genotype IA (n = 50) and IB (n = 11). The prevalent strain circulating in 2015 in both bivalves and waters was the IA strain responsible for the outbreak occurring around the same time in the Naples area. This variant was no longer identified in subsequent years (2017-2018) when, instead, appeared two of the IA variants of the multistate outbreak affecting men who have sex with men (MSM), VRD_521_2016, and RIVM-HAV16-090, with the former prevailing in both shellfish and water environments. HAV IB isolates were detected over the years in shellfish and in water matrices, but not in clinical samples, suggesting that this genotype had been circulating silently. An integrated surveillance system (environment/food/clinical cases) can be a useful tool to monitor changes in viral variants in the population, as well as an early warning system.


Assuntos
Microbiologia Ambiental , Vírus da Hepatite A/classificação , Hepatite A/epidemiologia , Hepatite A/virologia , Animais , Monitoramento Biológico , Bivalves , Monitoramento Ambiental , Genótipo , Geografia , Vírus da Hepatite A/genética , Humanos , Filogenia , Vigilância em Saúde Pública , RNA Viral , Água do Mar/virologia , Esgotos/virologia , Frutos do Mar/virologia
6.
Nat Microbiol ; 5(9): 1069-1078, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32451473

RESUMO

The Picornaviridae are a diverse family of positive-strand RNA viruses that includes numerous human and veterinary pathogens1. Among these, hepatitis A virus (HAV), a common cause of acute hepatitis in humans, is unique in that it is hepatotropic and is released from hepatocytes without lysis in small vesicles that resemble exosomes2,3. These quasi-enveloped virions are infectious and are the only form of virus that can be detected in the blood during acute infection2. By contrast, non-enveloped naked virions are shed in faeces and stripped of membranes by bile salts during passage through the bile ducts to the gut4. How these two distinct types of infectious hepatoviruses enter cells to initiate infection is unclear. Here, we describe a genome-wide forward screen that shows that glucosylceramide synthase and other components of the ganglioside synthetic pathway are crucial host factors that are required for cellular entry by hepatoviruses. We show that gangliosides-preferentially disialogangliosides-function as essential endolysosome receptors that are required for infection by both naked and quasi-enveloped virions. In the absence of gangliosides, both virion types are efficiently internalized through endocytosis, but capsids fail to uncoat and accumulate within LAMP1+ endolysosomes. Gangliosides relieve this block, binding to the capsid at low pH and facilitating a late step in entry involving uncoating and delivery of the RNA genome to the cytoplasm. These results reveal an atypical cellular entry pathway for hepatoviruses that is unique among picornaviruses.


Assuntos
Endossomos/metabolismo , Gangliosídeos/genética , Gangliosídeos/metabolismo , Vírus da Hepatite A/genética , Vírus da Hepatite A/metabolismo , Capsídeo/metabolismo , Proteínas do Capsídeo/metabolismo , Linhagem Celular , Endocitose , Exossomos , Técnicas de Inativação de Genes , Genoma Viral , Células HeLa , Hepatócitos/metabolismo , Humanos , Glicoproteínas de Membrana Associadas ao Lisossomo , Lisossomos/metabolismo , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Vírion/metabolismo , Internalização do Vírus
7.
PLoS One ; 15(5): e0234010, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32470049

RESUMO

In Europe HAV infection occurs mainly among specific risk groups, such as consumers of specific food. Sexual transmission of HAV has been demonstrated, particularly among Men-Who-Have-Sex-With-Men (MSM), causing MSM-specific outbreaksin Europe. Here we report a molecular epidemiologic and phylodynamic analysis on HAV sequences in Lazio (central Italy)to identify genetic background and the phylogenetic relations, and test the HAV infection dynamics during a large outbreak through phylodynamic model.Among all HAV sequences found during 2013-2018 in Lazio, low genetic diversity was observed in HAV population in 2016 and 2017, along with high frequenciesVRD_521_2016and RIVM-HAV16-090, suggesting a large expansion event of viral population. The initial expansion of both VRD_521_2016 and RIVM-HAV16-090 clusters dated back to 2012 (95% HPD:2006-2015). During the2016-2017outbreak in Lazio region, the Re peaked around mid-2016, with a value of 1.73 (95% HPD: 1.03-2.37), consistent with incidence trend of AHA cases in Lazio between 2016 and mid-2017. This study showed the magnitude of HAV outbreak in Lazio during 2016-2017, demonstrating the epidemic continuity to MSM-specific outbreak in Europe. The HAV dataset is available on interactive phylodynamic platform https://nextstrain.org to real-time update of future outbreaks.


Assuntos
Surtos de Doenças , Vírus da Hepatite A/classificação , Hepatite A/epidemiologia , Hepatite A/virologia , Homossexualidade Masculina , Filogenia , Sequência de Bases , Número Básico de Reprodução , Teorema de Bayes , Variação Genética , Genótipo , Vírus da Hepatite A/genética , Humanos , Itália/epidemiologia , Funções Verossimilhança , Masculino , Dinâmica Populacional
8.
Int J Food Microbiol ; 315: 108386, 2020 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-31707276

RESUMO

The NucliSENS MiniMAG (Minimag) system from bioMérieux is widely used for extraction of viral RNA from oysters and is included as informative material in the ISO method for quantification of hepatitis A virus (HAV) and norovirus genogroups I and II (GI and GII) in food (ISO 15216-1:2017). However, the system is no longer on sale within the EU and alternative methods are therefore needed. We optimised and evaluated an automated benchtop system for extraction of viral RNA from oysters artificially contaminated with HAV, norovirus GI, norovirus GII and mengovirus, using the same reagents and a similar protocol as with the Minimag method. Using the automated system instead of Minimag increased measured viral concentration by on average 1.3 times, suggesting that the automated system extracts viral RNA more efficiently than Minimag. A drawback with the automated system was that it displayed higher variability in measured concentration for mengovirus. The median viral recovery was 17%, 37%, 44% and 41% for samples extracted with the automated system and 15%, 27%, 34% and 23% for samples extracted with Minimag for HAV, norovirus GI, norovirus GII and mengovirus, respectively. All samples displayed <75% inhibition in RT-qPCR when extracted with the automated system or Minimag. Together, these results suggest that the automated system can be a suitable alternative to Minimag in analysis of HAV, norovirus GI and norovirus GII in oysters. However, verification using naturally contaminated oysters is needed before it can be used for food safety control purposes.


Assuntos
Vírus da Hepatite A/genética , Mengovirus/genética , Norovirus/genética , Ostreidae/virologia , RNA Viral/análise , Animais , Inocuidade dos Alimentos , RNA Viral/química , Reação em Cadeia da Polimerase em Tempo Real/métodos
9.
Jpn J Infect Dis ; 73(2): 89-95, 2020 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-31666497

RESUMO

Since 2017, hepatitis A virus (HAV) infection has been an epidemic among men who have sex with men (MSM) in Japan. We have come across 11 MSM patients with hepatitis A who were also infected with HIV. In 1999-2000, we came across 5 HIV-infected patients with hepatitis A. Since the conditions of current HIV-infected patients have changed owing to the recent progress in anti-HIV therapies, we compared clinical features of hepatitis A between patients in 2017-2018 and those in 1999-2000. By comparing the background characteristics of the patients, we found that the CD4/CD8 ratio was significantly higher in the 2017-2018 group. After the onset of hepatitis, peak levels of hepatic transaminases were found to be higher in the 2017-2018 group, suggesting severe hepatocellular damage. In contrast, neither the peak level of total bilirubin nor the nadir of prothrombin time was significantly different among the 2 groups. We also analyzed the HAV genome derived from some of the recently infected patients, and found that the HAV strains were almost the same among these patients; slight differences were observed from the previously identified strain. Thus, we concluded that the recovery of immunity by recent anti-HIV therapies may result in more severe hepatocellular damages and differences in clinical features.


Assuntos
Infecções por HIV/epidemiologia , Hepatite A/epidemiologia , Adulto , Antirretrovirais/efeitos adversos , Antirretrovirais/uso terapêutico , Contagem de Linfócito CD4 , Linfócitos T CD8-Positivos , Cidades/epidemiologia , Coinfecção/virologia , Genoma Viral , Infecções por HIV/virologia , Hepatite A/imunologia , Anticorpos Anti-Hepatite A/sangue , Vírus da Hepatite A/genética , Vírus da Hepatite A/imunologia , Homossexualidade Masculina , Humanos , Japão/epidemiologia , Fígado/efeitos dos fármacos , Fígado/patologia , Fígado/virologia , Masculino , Pessoa de Meia-Idade , Minorias Sexuais e de Gênero
10.
Int J Food Microbiol ; 311: 108349, 2019 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-31634688

RESUMO

Food-borne viral infections are caused mainly by noroviruses (NoV) and the hepatitis A virus (HAV), which respectively cause gastroenteritis and hepatitis. Various foods have been implicated in viral outbreaks, including vegetables that are consumed in a variety of forms, often with salad dressing. NF EN ISO procedures (15216-1:2017) propose standard methods for quantifying NoV and HAV in high-risk food categories, such as vegetables, based on viral elution and PEG concentration methods, but these methods are not suitable for composite meals like salads dressed with oily, fatty or emulsified food ingredients. The development of sensitive and reliable techniques for the detection of viruses in these products is therefore needed to ensure the safety of these products. The aim of this study was to develop an RT-qPCR based method for the detection and quantification of NoV and HAV in various vegetables with different dressings. Three methods for recovering NoV and HAV from artificially contaminated dressed vegetables were evaluated. The selected method was based on the use of Trizol reagent and, according to the type of dressing, the limit of detection ranged from 104 to 106 genome copies/g for NoV and from 102 to 103 PFU/g for HAV. The described method can be applied for detecting NoV and HAV in food containing salad dressing for routine diagnosis needs.


Assuntos
Contaminação de Alimentos/análise , Microbiologia de Alimentos/métodos , Vírus da Hepatite A/isolamento & purificação , Norovirus/isolamento & purificação , Verduras/virologia , Doenças Transmitidas por Alimentos/prevenção & controle , Doenças Transmitidas por Alimentos/virologia , Gastroenterite/prevenção & controle , Gastroenterite/virologia , Genoma Viral/genética , Vírus da Hepatite A/genética , Humanos , Norovirus/genética , RNA Viral/análise , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa
11.
J Vet Med Sci ; 81(10): 1533-1539, 2019 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-31406032

RESUMO

Duck hepatitis A virus (DHAV) infection is characterized by severe hepatitis. In recent years, DHAV-A has become widespread in Asia and has led to economic losses. Conventional methods of DHAV-A detection must often be performed in the laboratory with inconvenience equipment. We have developed a rapid reverse transcription insulated isothermal (RT-iiPCR) technique for the on-site detection of DHAV-A based on the POCKITTM system in a convenient minitype device. We optimized the PCR primers and probes for the amplification of the DHAV-A 3C/3D genes, and successfully amplified a specific fragment of DHAV-A, but no fragment from 18 other duck pathogens. The limit of detection for viral RNA was 49 copies per reaction, and the sensitivity and specificity were each 100% in the analysis of 60 liver samples. By comparison, the sensitivities of RT-iiPCR was comparable in sensitivity to existing rRT-PCR. Furthermore, the RT-iiPCR results were 98.3% in agreement with those of the rRT-PCR, with a kappa value of 0.938. In conclusion, this new method not only offers a higher sensitivity and specificity than existing techniques, but also time-saving and better suited to field diagnoses because device is portable.


Assuntos
Patos , Hepatite A/veterinária , Vírus da Hepatite do Pato/isolamento & purificação , Fígado/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Animais , China/epidemiologia , Hepatite A/diagnóstico , Vírus da Hepatite A/genética , Vírus da Hepatite do Pato/genética , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos
12.
Food Environ Virol ; 11(4): 393-399, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31420848

RESUMO

Enteroviruses (EVs) and Hepatitis A Viruses (HAVs) are human pathogens with a wide spectrum of clinical manifestations. The monitoring of sewage samples enables to monitor the EVs and HAVs in circulation among the general population and recognize possible outbreaks. This study focused on the molecular characterization and phylogenetic analysis of the EVs and HAVs identified in 33 sewage samples collected every 15 days at the influent of a wastewater treatment plant located in Northern Italy from March to October 2016. According to the results of the molecular characterization, the most frequently identified viruses were Echovirus 6 (E-6), E-11 and HAV-IA. The phylogenetic analyses indicated the rapid genetic evolution of E-6 and E-1; noteworthy, most E-11 strains clustered with a strain isolated from a clinical sample collected in the same geographical area over the same period by our laboratory. Most of the HAV strains detected clustered with epidemic HAV-IA strains identified during the European hepatitis A outbreak that occurred in 2016-2017 affecting men who have sex with men (MSM). The detection of environmental HAV strains before and at the beginning of its spread amongst humans demonstrated that this outbreak could have been predicted by monitoring sewage samples. Moreover, conducting a genetic comparison between the HAV and EV strains identified in sewage and clinical samples may improve knowledge of viral epidemiology. EV and HAV molecular environmental surveillance may prove useful for identifying viral circulation and for issuing early warning alerts on possible outbreaks among the human population.


Assuntos
Infecções por Enterovirus/virologia , Enterovirus/genética , Vírus da Hepatite A/genética , Hepatite A/virologia , Filogenia , Esgotos/virologia , Enterovirus/classificação , Enterovirus/isolamento & purificação , Infecções por Enterovirus/epidemiologia , Hepatite A/epidemiologia , Vírus da Hepatite A/classificação , Vírus da Hepatite A/isolamento & purificação , Humanos , Itália/epidemiologia
13.
Food Microbiol ; 84: 103257, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31421763

RESUMO

The viability of murine norovirus (MNV-1), bovine rotavirus (boRV), and hepatitis A virus (HAV) was evaluated at 21 °C, 4 °C, and -20 °C on stainless steel surfaces, in bottled water, and on blueberries for up to 21 days. After 14 days of incubation at 21 °C on stainless steel, a viability loss >4 log for MNV-1, >8 log for boRV, and >1 log for HAV was observed. Losses were observed for MNV-1 (>1 log) and HAV (>2 log) incubated in water at 21 °C for 21 days. No significant loss was detected for MNV-1 and HAV at 4 °C and -20 °C and for boRV at 21 °C, 4 °C, and -20 °C. On blueberries incubated at 4 °C and -20 °C, they all maintained their infectivity. After 7 days at 21 °C, a loss >2 log, a loss of 3 log, and no loss were observed for boRV, MNV-1, and HAV, respectively. After RNase pretreatment, the detection of extracted RNA from infectious and noninfectious samples suggested the protection of RNA inside the capsid. Even though they all are enteric viruses, their persistence varied with temperature and the nature of the commodity. It is therefore important to use more than one viral surrogate, during inactivation treatments or implementation of control measures.


Assuntos
Mirtilos Azuis (Planta)/virologia , Água Potável/microbiologia , Vírus da Hepatite A/isolamento & purificação , Norovirus/isolamento & purificação , Rotavirus/isolamento & purificação , Aço Inoxidável/análise , Inativação de Vírus , Animais , Bovinos , Linhagem Celular , Desinfecção , Contaminação de Alimentos/análise , Vírus da Hepatite A/genética , Camundongos , Norovirus/genética , RNA Viral , Rotavirus/genética , Temperatura
14.
Epidemiol Infect ; 147: e210, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-31364560

RESUMO

India is experiencing a substantial decrease in early childhood exposure to hepatitis A virus (HAV). Kerala has experienced several hepatitis A outbreaks in young adults/adults in the recent past. The current hepatitis outbreak occurred in Nellikuzhi, Kerala state, India in December 2016. Investigation was carried by preparing a line list of suspected hepatitis cases. The blood and stool samples collected from patients were tested for anti-HAV/anti-Hepatitis E virus (HEV) immunoglobulin (IgM) antibodies and RNA respectively. A total of 562 suspected hepatitis cases were reported during the outbreak. Along with the first case (35 years, male), 86.1% (484/562) of the cases gave history of consuming food/water/cold drinks from one restaurant. Anti-HAV IgM positivity was 74.5% (73/98) in tested samples and amongst the positives, 81% were adults/young adults and adolescents. None of the samples tested positive for anti-HEV IgM. There were three HAV associated deaths without any co-morbidity. Sequence analysis of HAV RNA positive stool samples showed the presence of genotype IIIA HAV. The suspected source of the infection was a private well situated in the premise of a restaurant. Considering increasing HAV naive population in Kerala, there is a need to introduce hepatitis A vaccine in high-risk age groups.


Assuntos
Transmissão de Doença Infecciosa , Doenças Transmitidas por Alimentos/epidemiologia , Vírus da Hepatite A/isolamento & purificação , Hepatite A/epidemiologia , Hepatite A/transmissão , Restaurantes , População Rural , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Fezes/virologia , Feminino , Genótipo , Anticorpos Anti-Hepatite A/sangue , Vírus da Hepatite A/classificação , Vírus da Hepatite A/genética , Vírus da Hepatite A/imunologia , Humanos , Imunoglobulina M/sangue , Índia/epidemiologia , Masculino , Pessoa de Meia-Idade , RNA Viral/análise , RNA Viral/genética , Análise de Sobrevida , Adulto Jovem
15.
Genome Biol Evol ; 11(9): 2439-2456, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31290967

RESUMO

Hepatoviruses show an intriguing deviated codon usage, suggesting an evolutionary signature. Abundant and rare codons in the cellular genome are scarce in the human hepatitis A virus (HAV) genome, while intermediately abundant host codons are abundant in the virus. Genotype-phenotype maps, or fitness landscapes, are a means of representing a genotype position in sequence space and uncovering how genotype relates to phenotype and fitness. Using genotype-phenotype maps of the translation efficiency, we have shown the critical role of the HAV capsid codon composition in regulating translation and determining its robustness. Adaptation to an environmental perturbation such as the artificial induction of cellular shutoff-not naturally occurring in HAV infection-involved movements in the sequence space and dramatic changes of the translation efficiency. Capsid rare codons, including abundant and rare codons of the cellular genome, slowed down the translation efficiency in conditions of no cellular shutoff. In contrast, rare capsid codons that are abundant in the cellular genome were efficiently translated in conditions of shutoff. Capsid regions very rich in slowly translated codons adapt to shutoff through sequence space movements from positions with highly robust translation to others with diminished translation robustness. These movements paralleled decreases of the capsid physical and biological robustness, and resulted in the diversification of capsid phenotypes. The deviated codon usage of extant hepatoviruses compared with that of their hosts may suggest the occurrence of a virus ancestor with an optimized codon usage with respect to an unknown ancient host.


Assuntos
Proteínas do Capsídeo/genética , Vírus da Hepatite A/genética , Vírus da Hepatite A/fisiologia , Elongação Traducional da Cadeia Peptídica , Adaptação Fisiológica , Proteínas do Capsídeo/metabolismo , Códon , Humanos , Mutação , Biossíntese de Proteínas , Dobramento de Proteína , RNA de Transferência/metabolismo
16.
Emerg Infect Dis ; 25(8): 1594-1596, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31310213

RESUMO

We implemented subgenomic and whole-genome sequencing to support the investigation of a large hepatitis A virus outbreak among persons experiencing homelessness, users of illicit drugs, or both in California, USA, during 2017-2018. Genotyping data helped confirm case-patients, track chains of transmission, and monitor the effectiveness of public health control measures.


Assuntos
Vírus da Hepatite A/classificação , Vírus da Hepatite A/genética , Hepatite A/epidemiologia , Hepatite A/virologia , Tipagem Molecular , California/epidemiologia , Surtos de Doenças , Genótipo , Hepatite A/história , Hepatite A/transmissão , Vírus da Hepatite A/imunologia , Vírus da Hepatite A/isolamento & purificação , História do Século XXI , Humanos , Vigilância em Saúde Pública , Proteínas Virais/genética , Sequenciamento Completo do Genoma
17.
Euro Surveill ; 24(28)2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31311618

RESUMO

IntroductionSequence-based typing of hepatitis A virus (HAV) is important for outbreak detection, investigation and surveillance. In 2013, sequencing was central to resolving a large European Union (EU)-wide outbreak related to frozen berries. However, as the sequenced HAV genome regions were only partly comparable between countries, results were not always conclusive.AimThe objective was to gather information on HAV surveillance and sequencing in EU/European Economic Area (EEA) countries to find ways to harmonise their procedures, for improvement of cross-border outbreak responses.MethodsIn 2014, the European Centre for Disease Prevention and Control (ECDC) conducted a survey on HAV surveillance practices in EU/EEA countries. The survey enquired whether a referral system for confirming primary diagnostics of hepatitis A existed as well as a central collection/storage of hepatitis A cases' samples for typing. Questions on HAV sequencing procedures were also asked. Based on the results, an expert consultation proposed harmonised procedures for cross-border outbreak response, in particular regarding sequencing. In 2016, a follow-up survey assessed uptake of suggested methods.ResultsOf 31 EU/EEA countries, 23 (2014) and 27 (2016) participated. Numbers of countries with central collection and storage of HAV positive samples and of those performing sequencing increased from 12 to 15 and 12 to 14 respectively in 2016, with all countries typing an overlapping fragment of 218 nt. However, variation existed in the sequenced genomic regions and their lengths.ConclusionsWhile HAV sequences in EU/EEA countries are comparable for surveillance, collaboration in sharing and comparing these can be further strengthened.


Assuntos
Surtos de Doenças/prevenção & controle , Vírus da Hepatite A/isolamento & purificação , Hepatite A/diagnóstico , Tipagem Molecular/métodos , Vigilância da População/métodos , Sequenciamento Completo do Genoma/métodos , Europa (Continente)/epidemiologia , União Europeia , Hepatite A/epidemiologia , Vírus da Hepatite A/genética , Humanos , RNA Viral/análise , Análise de Sequência de DNA
18.
J Clin Microbiol ; 57(9)2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31217273

RESUMO

Laboratory surveillance plays an important role in the detection and control of hepatitis A outbreaks and requires the application of rapid and accurate molecular diagnostic tools for hepatitis A virus (HAV) RNA detection, subgenotype identification, and sequence-based genotyping. We describe the development and validation of a triplex real-time, reverse transcription-PCR (triplex rRT-PCR) assay for the identification and discrimination of HAV subgenotypes IA, IB, and IIIA and a singleplex rRT-PCR assay designed to detect all HAV genotypes infecting humans. Overall, the accuracy, sensitivity, and specificity of the new assays were >97% for serum and plasma specimens collected during unrelated outbreaks of HAV in California and Michigan compared to a nested RT-PCR genotyping assay and the ISO 15216-1 rRT-PCR method for HAV detection. The new assays will permit the rapid detection of HAV RNA and discrimination among subgenotypes IA, IB, and IIIA in serum and plasma specimens, which will strengthen public health surveillance efforts for HAV outbreak detection and response.


Assuntos
Técnicas de Genotipagem/métodos , Vírus da Hepatite A/classificação , Vírus da Hepatite A/isolamento & purificação , Hepatite A/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , California/epidemiologia , Surtos de Doenças , Monitoramento Epidemiológico , Genótipo , Hepatite A/epidemiologia , Hepatite A/virologia , Vírus da Hepatite A/genética , Humanos , Michigan/epidemiologia , Epidemiologia Molecular/métodos , Sensibilidade e Especificidade
19.
J Clin Virol ; 117: 49-53, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31202104

RESUMO

BACKGROUND: During June-2016-May-2017, several outbreaks of HA were recorded in Europe, especially described in MSM. In our area since July-2017, an increase of hepatitis A (HA) notification was reported. OBJECTIVE: In order to understand the unusual increase of cases occurred in the central region of Argentina, the aim of this study was to describe, characterize and contextualize epidemiologically the HA outbreak occurred this area, until April2018. STUDY DESIGN: HA cases (positive anti-HAV IgM) obtained from the calendar week 29/2017 in which the first case of MSM was recognized were included in our study. HAV RNA detection and molecular characterization was performed from serum samples and/or stool by RT - PCR of VP1/2A genomic region (360bp). RESULTS: Of the 32 cases notified, 87.5% of them were unvaccinated men and 69.6% were MSM (mean age 31.9 years). All MSM associated HAV sequences were genotyped as IA, and clustered with the VRD 521-2016 strain, responsible of causing outbreaks mostly in MSM in Europe since mid-2016. CONCLUSION: As a consequence of the implementation of immunization in children, and the improvement in socio-economic, hygienic and sanitation factors, young adults are becoming increasingly susceptible to HAV infections. Here we add evidence in South America to the HA outbreaks described worldwide among young MSM, demonstrating the need to reinforce official policy of vaccination, in this group and adjust epidemiological surveillance, catch-up vaccination for adolescents, young adults and immunosuppressed patients.


Assuntos
Surtos de Doenças , Vírus da Hepatite A/classificação , Hepatite A/epidemiologia , Homossexualidade Masculina/estatística & dados numéricos , Adulto , Fatores Etários , Argentina/epidemiologia , Europa (Continente)/epidemiologia , Feminino , Genótipo , Hepatite A/diagnóstico , Vírus da Hepatite A/genética , Vírus da Hepatite A/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Filogenia , Caracteres Sexuais , Proteínas Estruturais Virais/genética , Adulto Jovem
20.
Int J Infect Dis ; 86: 12-14, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31238154

RESUMO

OBJECTIVES: To investigate the prevalence of anti-HAV and HEV markers in order to better understand spread of these two viruses among adults in Rwanda. METHODS: Samples from 1045 and 1133 blood donors, healthy adults and liver disease patients were analysed for anti-HAV IgG and HEV markers respectively. RESULTS: Anti-HAV was present in 96.9% (1013/1045), with proportions of immune persons increasing with age. HEV infection markers were detected in 11.9% (135/1133) without differences between the three categories. Seven persons had low levels of HEV RNA including four blood donors but none of the HEV strains could be sequenced. The highest prevalence of HEV markers was in farmers and persons from the Southern (17.3%) and Western regions (18.6%), which have the national highest density of pigs. This may indicate that pigs constitute an important source of HEV infection for humans in Rwanda. CONCLUSION: HAV remains highly endemic in Rwanda, but there may now be a decline of exposure during childhood. HEV is also endemic in Rwanda, but has a moderate spread and may be transmitted by blood transfusion. Based on the geographical and occupational differences in HEV prevalence, a possible zoonotic transmission from pigs should be further explored.


Assuntos
Vírus da Hepatite A/fisiologia , Hepatite A/epidemiologia , Vírus da Hepatite E/fisiologia , Hepatite E/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Doadores de Sangue/estatística & dados numéricos , Feminino , Hepatite A/sangue , Hepatite A/transmissão , Hepatite A/virologia , Vírus da Hepatite A/genética , Vírus da Hepatite A/isolamento & purificação , Anticorpos Anti-Hepatite/sangue , Hepatite E/sangue , Hepatite E/transmissão , Hepatite E/virologia , Vírus da Hepatite E/genética , Vírus da Hepatite E/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Ruanda/epidemiologia , Estudos Soroepidemiológicos , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/virologia , Adulto Jovem , Zoonoses/sangue , Zoonoses/epidemiologia , Zoonoses/virologia
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