Development and clinical validation of a pangenotypic PCR-based assay for the detection and quantification of hepatitis E virus (Orthopevirus A genus)

The objective of this study was to design a pangenotypic PCR-based assay for the detection and quantification of hepatitis E virus (HEV) RNA from across the entire spectrum of described genotypes belonging to the Orthohepevirus A genus. The optimal conditions and the performance of the assay were determined by testing the WHO standard strain (6219/10) and the WHO HEV panel (8578/13). Similarly, performance comparisons were made with two commercial assays (Real Star HEV RT-PCR 2.0 and ampliCube HEV 2.0 Quant) to detect HEV RNA at concentrations below 1,000 IU/mL with viral strains from the WHO and to test samples from 54 patients with acute hepatitis. The assay presented in this study was able to detect the entire spectrum of described genotypes belonging to the Orthohepevirus A genus, demonstrating a better performance than both commercial kits. This procedure may represent a significant improvement in the molecular diagnosis of HEV infection.

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Hepatitis E Virus Occurrence in Pigs Slaughtered in Italy

Animals ◽

10.3390/ani11020277 ◽

2021 ◽

Vol 11 (2) ◽

pp. 277

Author(s):

Eleonora Chelli ◽

Elisabetta Suffredini ◽

Paola De Santis ◽

Dario De Medici ◽

Santina Di Bella ◽

...

Keyword(s):

Hepatitis E Virus ◽

Phylogenetic Analyses ◽

Hepatitis E ◽

Rt Pcr ◽

High Seroprevalence ◽

Genotype 3 ◽

Sporadic Cases ◽

Wild Boar Meat ◽

Small Clusters ◽

Frequent Exposure

In Europe, foodborne transmission has been clearly associated to sporadic cases and small clusters of hepatitis E in humans linked to the consumption of contaminated pig liver sausages, raw venison, or undercooked wild boar meat. In Europe, zoonotic HEV-genotype 3 strains are widespread in pig farms but little information is available on the prevalence of HEV positive pigs at slaughterhouse. In the present study, the prevalence of HEV-RNA positive pigs was assessed on 585 animals from 4 abattoirs located across Italy. Twenty-one pigs (3.6%) tested positive for HEV in either feces or liver by real-time RT-PCR. In these 21 pigs, eight diaphragm muscles resulted positive for HEV-RNA. Among animals collected in one abattoir, 4 out of 91 plasma tested positive for HEV-RNA. ELISA tests for the detection of total antibodies against HEV showed a high seroprevalence (76.8%), confirming the frequent exposure of pigs to the virus. The phylogenetic analyses conducted on sequences of both ORF1 and ORF2 fragments, shows the circulation of HEV-3c and of a novel unclassified subtype. This study provides information on HEV occurrence in pigs at the slaughterhouse, confirming that muscles are rarely contaminated by HEV-RNA compared to liver, which is the most frequently positive for HEV.

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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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Detection and characterisation of hepatitis E virus in naturally infected swine in Croatia

Acta Veterinaria Hungarica ◽

10.1556/avet.2013.031 ◽

2013 ◽

Vol 61 (4) ◽

pp. 517-528 ◽

Cited By ~ 8

Author(s):

Zoran Lipej ◽

Dinko Novosel ◽

Lea Vojta ◽

Besi Roić ◽

Miljenko Šimpraga ◽

...

Keyword(s):

Hepatitis E Virus ◽

Age Groups ◽

Hepatitis E ◽

Health Concern ◽

Rt Pcr ◽

Animal Reservoir ◽

Naturally Occurring ◽

High Prevalence ◽

Swine Herds ◽

Bile Samples

Hepatitis E is a viral zoonotic disease infecting swine worldwide. Since pigs represent a likely animal reservoir for the hepatitis E virus, the epidemiology of naturally occurring hepatitis E was investigated in Croatian swine herds. Nearly all tested animals were seropositive for antibodies against the hepatitis E virus (55/60, 91.7%). Active infection was detected in all age groups by RT-PCR of viral RNA in serum (8/60, 13.3%) and bile samples (3/37, 8.1%), which was further confirmed by histopathological findings of characteristic lesions in the livers of the infected animals. Three new strains of hepatitis E virus were isolated from Croatian pig herds. Phylogenetic analysis using median-joining networks clustered those Croatian strains with isolates from various parts of the world, indicating their likely origin in international trade. Similarity to human isolates implies a zoonotic potential of Croatian strains, which raises a public health concern, especially in the light of the high prevalence of hepatitis E in the herds studied.

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Hepatitis E Virus Quasispecies and the Outcome of Acute Hepatitis E in Solid-Organ Transplant Patients

Journal of Virology ◽

10.1128/jvi.01003-12 ◽

2012 ◽

Vol 86 (18) ◽

pp. 10006-10014 ◽

Cited By ~ 71

Author(s):

Sebastien Lhomme ◽

Florence Abravanel ◽

Martine Dubois ◽

Karine Sandres-Saune ◽

Lionel Rostaing ◽

...

Keyword(s):

Liver Fibrosis ◽

Acute Hepatitis ◽

Hepatitis E Virus ◽

Organ Transplant ◽

Hepatitis E ◽

Solid Organ Transplant ◽

First Year ◽

Solid Organ ◽

Immunocompromised Patients ◽

Serum Concentrations

Hepatitis E virus (HEV) infections are responsible for chronic hepatitis in immunocompromised patients, and this can evolve to cirrhosis. Like all RNA viruses, HEV exists as a mixture of heterogeneous viruses defining quasispecies. The relationship between the genetic heterogeneity described as a quasispecies, cytokine secretion, and the outcome of acute hepatitis in immunocompromised patients remains to be elucidated. We cloned and sequenced the region encoding the M and P capsid domains of HEV from eight solid-organ transplant (SOT) patients with acute HEV infection who subsequently cleared the virus and from eight SOT patients whose infection became chronic. We analyzed the cytokines and chemokines in the sera of these SOT patients by multianalyte profiling. The nucleotide sequence entropy and genetic distances were greater in patients whose infections became chronic. A lowerKa/Ksratio was associated with the persistence of HEV. The patients who developed chronic infection had lower serum concentrations of interleukin-1 (IL-1) receptor antagonist and soluble IL-2 receptor. Increased concentrations of the chemokines implicated in leukocyte recruitment to the liver were associated with persistent infection. Those patients with chronic HEV infection and progressing liver fibrosis had less quasispecies diversification during the first year than patients without liver fibrosis progression. Great quasispecies heterogeneity, a weak inflammatory response, and high serum concentrations of the chemokines involved in leukocyte recruitment to the liver in the acute phase were associated with persistent HEV infection. Slow quasispecies diversification during the first year was associated with rapidly developing liver fibrosis.

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