Review: Approaches to the Viral Extraction, Detection, and Identification of Hepatitis Viruses, HAV and HEV, in Foods

Abstract Although the incidence rate of hepatitis A virus (HAV) infection has been on the decline in developed countries, in part due to immunization availability, high profile outbreaks continue to be reported. Hepatitis E virus has been recognized as an emerging pathogen in industrialized countries. While associated with waterborne illnesses, particularly in undeveloped countries, several animal species have been identified as reservoirs for the virus. Consequently, the potential of zoonotic transmission exists as a function of the consumption of infected animals. In this review we provide a comparative overview of these two virus species with regard to their known virus properties, discuss extraction methodologies, and describe some basic principles and methodology applied toward the isolation of these viruses (as particles or their isolated genomes) from food commodities. We also discuss the challenges that remain as experimental hurdles to extraction of such viruses from food. As HAV has been the most extensively studied with regard to virus detection in foods, it often serves as a model virus for current and future development of sample preparation methodology for foodborne virus detection. Lastly, we discuss the application and role of current and developing technologies in the post-extraction detection and identification of these viruses from foods.

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Seroprevalence for hepatitis E virus (HEV) infection among volunteer blood donors of the Regional Blood Bank of Londrina, State of Paraná , Brazil

Revista do Instituto de Medicina Tropical de São Paulo ◽

10.1590/s0036-46652006000200006 ◽

2006 ◽

Vol 48 (2) ◽

pp. 87-92 ◽

Cited By ~ 30

Author(s):

André Luiz Bortoliero ◽

Ana Maria Bonametti ◽

Helena Kaminami Morimoto ◽

Tiemi Matsuo ◽

Edna Maria Vissoci Reiche

Keyword(s):

Urban Areas ◽

Hepatitis E Virus ◽

Blood Donors ◽

Hepatitis A Virus ◽

Hepatitis E ◽

Developed Countries ◽

Blood Bank ◽

Igg Antibodies ◽

Serum Samples ◽

Sociodemographic Variables

A cross-sectional study was carried out among 996 volunteer blood donors enrolled from May 1999 to December 1999 to determine the seroprevalence of hepatitis E virus (HEV) infection among volunteer blood donors of the Regional Blood Bank of Londrina, State of Paraná, Brazil, and to evaluate whether the rate of seroprevalence of IgG anti-HEV antibodies is associated with sociodemographic variables and with seropositivity for hepatitis A virus (HAV) infection. All participants answered the questionnaire regarding the sociodemographic characterisitcs. Serum samples were tested for IgG antibodies to HEV (anti-HEV) by an enzyme linked immunoassay (ELISA). All serum samples positive for anti-HEV IgG and 237 serum samples negative for anti-HEV were also assayed for IgG anti-HAV antibodies by ELISA. Anti-HEV IgG was confirmed in 23/996 samples, resulting in a seroprevalence of 2.3% for HEV infection, similar to previous results obtained in developed countries. No significant association was found between the presence of anti-HEV IgG antibodies and the sociodemographic variables including gender, age, educational level, rural or urban areas, source of water, and sewer system (p > 0.05). Also, no association with seropositivity for anti-HAV IgG antibodies was observed (p > 0.05). Although this study revealed a low seroprevalence of HEV infection in the population evaluated, the results showed that this virus is circulating among the population from Londrina, South Brazil, and point out the need of further studies to define the clinical and epidemiological importance of HEV infection and to identify additional risk factors involved in the epidemiology and pathogenesis of this infection in this population.

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Hepatitis E Virus in Industrialized Countries: The Silent Threat

BioMed Research International ◽

10.1155/2016/9838041 ◽

2016 ◽

Vol 2016 ◽

pp. 1-17 ◽

Cited By ~ 30

Author(s):

Pilar Clemente-Casares ◽

Carlota Ramos-Romero ◽

Eugenio Ramirez-Gonzalez ◽

Antonio Mas

Keyword(s):

Hepatitis E Virus ◽

Hepatitis E ◽

Developed Countries ◽

European Population ◽

Environmental Data ◽

Industrialized Countries ◽

Pork Products ◽

Weakest Link ◽

Imported Cases ◽

Serological Data

Hepatitis E virus (HEV) is the main cause of acute viral hepatitis worldwide. Its presence in developing countries has been documented for decades. Developed countries were supposed to be virus-free and initially only imported cases were detected in those areas. However, sporadic and autochthonous cases of HEV infection have been identified and studies reveal that the virus is worldwide spread. Chronic hepatitis and multiple extrahepatic manifestations have also been associated with HEV. We review the data from European countries, where human, animal, and environmental data have been collected since the 90s. In Europe, autochthonous HEV strains were first detected in the late 90s and early 2000s. Since then, serological data have shown that the virus infects quite frequently the European population and that some species, such as pigs, wild boars, and deer, are reservoirs. HEV strains can be isolated from environmental samples and reach the food chain, as shown by the detection of the virus in mussels and in contaminated pork products as sausages or meat. All these data highlight the need of studies directed to control the sources of HEV to protect immunocompromised individuals that seem the weakest link of the HEV epidemiology in industrialized regions.

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Identification of hepatitis a virus in cell cultures by solid phase immune electron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100142815 ◽

1986 ◽

Vol 44 ◽

pp. 238-239

Author(s):

C.D. Humphrey ◽

T.L. Cromeans ◽

E.H. Cook ◽

D.W. Bradley

Keyword(s):

Electron Microscopy ◽

Liquid Phase ◽

Cell Cultures ◽

Rapid Detection ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Solid Phase ◽

Immune Electron Microscopy ◽

Detection And Identification

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.

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Significant compartment‐specific impact of different RNA extraction methods and PCR assays on the sensitivity of Hepatitis E virus detection

Liver International ◽

10.1111/liv.14870 ◽

2021 ◽

Author(s):

Patrick Behrendt ◽

Birgit Bremer ◽

Daniel Todt ◽

Eike Steinmann ◽

Michael Peter Manns ◽

...

Keyword(s):

Hepatitis E Virus ◽

Rna Extraction ◽

Virus Detection ◽

Hepatitis E ◽

Extraction Methods ◽

Rna Extraction Methods ◽

Pcr Assays ◽

Specific Impact

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Identification and genetic diversity of hepatitis E virus in domestic swine from Slovakia

BMC Veterinary Research ◽

10.1186/s12917-021-02936-4 ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Anna Jackova ◽

Katarina Dudasova ◽

Slavomira Salamunova ◽

Rene Mandelik ◽

Jaroslav Novotny ◽

...

Keyword(s):

Genetic Diversity ◽

Hepatitis E Virus ◽

Hepatitis E ◽

Developed Countries ◽

European Countries ◽

Major Clade ◽

Information Gap ◽

Production Stages ◽

Suckling Piglets ◽

Domestic Swine

Abstract Background Hepatitis E virus (HEV) is agent causing hepatitis worldwide. Originally considered to be limited to developing countries, this virus was also detected in developed countries. In recent years an increasing number of reports indicate that farmed domestic pigs are widely infected with HEV in several European countries. The HEV status in Slovakia is still missing. Results In this study, the circulation of HEV among domestic swine in Slovakia and genetic diversity of the virus was studied. Overall HEV RNA was detected in 53/388 (13.7, 95% CI: 10.40–17.48%) pig rectal swabs in five production stages (age categories) with statistically significant differences among all the stages. The highest HEV prevalence was observed in weaners 24/81 (29.6, 95% CI: 19.99–40.81%) and then significantly declined in growers and fatteners. No HEV was detected in suckling piglets and sows. Twenty-eight partial sequences of ORF1 (242 bp) and seventeen of ORF2 (304 bp) were analysed. Phylogenetic analysis and p-distance comparisons confirmed in both ORFs that all Slovak HEV sequences belong to the genotype HEV-3, major clade 3abchij with higher identity to 3a and 3i subtypes. Three sequences were outside of all lastly updated HEV-3 subtypes. Conclusion This is the first report to fill the information gap about HEV infection in pigs in Slovakia. The results suggested a lower prevalence of HEV in Slovak pig farms than observed in other European countries. While most HEV isolates were typed as HEV-3 clade 3abchij, three sequences were unclassified.

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First Report of Hepatitis E Virus in Shellfish in Southeast Italy

Applied Sciences ◽

10.3390/app11010043 ◽

2020 ◽

Vol 11 (1) ◽

pp. 43

Author(s):

Gianfranco La Bella ◽

Maria Grazia Basanisi ◽

Gaia Nobili ◽

Valentina Terio ◽

Elisabetta Suffredini ◽

...

Keyword(s):

Real Time ◽

Hepatitis E Virus ◽

Potential Role ◽

Hepatitis E ◽

Developed Countries ◽

Rt Pcr ◽

Viral Pathogens ◽

Apulia Region ◽

Causative Agents

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

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Systematic Review of Hepatitis E Virus in Brazil: A One-Health Approach of the Human-Animal-Environment Triad

Animals ◽

10.3390/ani11082290 ◽

2021 ◽

Vol 11 (8) ◽

pp. 2290

Author(s):

Danny Franciele da Silva Dias Moraes ◽

João R. Mesquita ◽

Valéria Dutra ◽

Maria São José Nascimento

Keyword(s):

Systematic Review ◽

One Health ◽

Hepatitis E ◽

Published Data ◽

Surveillance Systems ◽

Industrialized Countries ◽

Genotype 3 ◽

The World ◽

Waterborne Transmission ◽

Sanitary Conditions

Brazil is the fifth largest country in the world with diverse socioeconomic and sanitary conditions, also being the fourth largest pig producer in the world. The aim of the present systematic review was to collect and summarize all HEV published data from Brazil (from 1995 to October 2020) performed in humans, animals, and the environment, in a One Health perspective. A total of 2173 papers were retrieved from five search databases (LILACs, Mendeley, PubMed, Scopus, and Web of Science) resulting in 71 eligible papers after application of exclusion/inclusion criteria. Data shows that HEV genotype 3 (HEV-3) was the only retrieved genotype in humans, animals, and environment in Brazil. The South region showed the highest human seroprevalence and also the highest pig density and industry, suggesting a zoonotic link. HEV-1 and 2 were not detected in Brazil, despite the low sanitary conditions of some regions. From the present review we infer that HEV epidemiology in Brazil is similar to that of industrialized countries (only HEV-3, swine reservoirs, no waterborne transmission, no association with low sanitary conditions). Hence, we alert for the implementation of HEV surveillance systems in swine and for the consideration of HEV in the diagnostic routine of acute and chronic hepatitis in humans.

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