scholarly journals An Optimized High-Throughput Neutralization Assay for Hepatitis E Virus (HEV) Involving Detection of Secreted Porf2

Viruses ◽  
2019 ◽  
Vol 11 (1) ◽  
pp. 64 ◽  
Author(s):  
Chang Liu ◽  
Wei Cai ◽  
Xin Yin ◽  
Zimin Tang ◽  
Guiping Wen ◽  
...  
Keyword(s):  
High Throughput ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Neutralization Assay ◽  
The Novel ◽  
Specific Antibodies ◽  
Neutralizing Activity ◽  
Elisa Kit ◽  
Neutralizing Capacity ◽  
Potential Threshold

Hepatitis E virus (HEV) is a common cause of acute hepatitis worldwide. Current methods for evaluating the neutralizing activity of HEV-specific antibodies include immunofluorescence focus assays (IFAs) and real-time PCR, which are insensitive and operationally complicated. Here, we developed a high-throughput neutralization assay by measuring secreted pORF2 levels using an HEV antigen enzyme-linked immunosorbent assay (ELISA) kit based on the highly replicating HEV genotype (gt) 3 strain Kernow. We evaluated the neutralizing activity of HEV-specific antibodies and the sera of vaccinated individuals (n = 15) by traditional IFA and the novel assay simultaneously. A linear regression analysis shows that there is a high degree of correlation between the two assays. Furthermore, the anti-HEV IgG levels exhibited moderate correlation with the neutralizing titers of the sera of vaccinated individuals, indicating that immunization with gt 1 can protect against gt 3 Kernow infection. We then determined specificity of the novel assay and the potential threshold of neutralizing capacity using anti-HEV IgG positive sera (n = 27) and anti-HEV IgG negative sera (n = 23). The neutralizing capacity of anti-HEV IgG positive sera was significantly stronger than that of anti-HEV IgG negative. In addition, ROC curve analysis shows that the potential threshold of neutralizing capacity of sera was 8.07, and the sensitivity and specificity of the novel assay was 88.6% and 100%, respectively. Our results suggest that the neutralization assay using the antigen ELISA kit could be a useful tool for HEV clinical research.

scholarly journals Serological evidence of Hepatitis E Virus infection in Antioquia, Colombia slaughtered pigs

2015 ◽  
pp. 4602-4613 ◽  
Author(s):  
Jorge Forero D ◽  
Cristian Gutiérrez V ◽  
Jaime Parra S ◽  
Guillermo Correa L ◽  
Berardo Rodríguez ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Igg Antibodies ◽  
Serological Evidence ◽  
Blood Samples ◽  
Specific Antibodies ◽  
Elisa Kit ◽  
Igm Antibodies ◽  
Slaughter Age ◽  
Slaughtered Pigs

ABSTRACTObjective. To detect the presence of specific antibodies against Hepatitis E virus (HEV) in pigs slaughtered in Antioquia, the department where the greatest amount of pork is produced and consumed in Colombia. Materials and methods. Between September 2011 and May 2012, blood samples from pigs were obtained in five slaughterhouses of Antioquia, four of them located in the Aburrá Valley subregion and other located in northern subregión. Serum were evaluated with a commercial ELISA kit for diagnosing HEV in humans but adapted to detect IgG and IgM antibodies in pigs. Results. A 100.0% seropositivity for IgG antibodies was found in 1000 samples evaluated, and 82.06% for IgM antibodies were found in 740 samples. Conclusions. These results indicate that pigs in slaughter age in Antioquia, and possibly in Colombia, have been exposed to HEV at some point in their production process and a high percentage of them can arrive to slaughterhouses with recent infection.RESUMENObjetivo. Detectar la presencia de anticuerpos específicos contra el virus de la Hepatitis E (HEV) en cerdos faenados en Antioquia, departamento donde se produce y consume la mayor cantidad de carne de cerdo en Colombia. Materiales y métodos. Entre septiembre de 2011 y mayo de 2012, se obtuvieron muestras de sangre de cerdos en cinco plantas de faenado, cuatro de ellas ubicadas en el Valle de Aburrá y una en la subregión Norte del departamento de Antioquia, las cuales fueron evaluadas mediante un kit de ELISA comercial para diagnóstico de HEV en humanos pero adaptado para la detección de anticuerpos tipo IgG e IgM en cerdos. Resultados. Se encontró una seropositividad de 100.0% para anticuerpos tipo IgG en 1000 muestras evaluadas y de 82.06% para anticuerpos tipo IgM en 740 muestras. Una muestra de heces positiva para la detección del genoma HEV es similar al genotipo 3 encontrada en Estados Unidos. Conclusiones. Estos resultados indican que los cerdos en edad de faenado en Antioquia y posiblemente en Colombia, han tenido exposición a HEV del, un virus zoonótico emergente a nivel mundial, en algún momento de su proceso productivo.

scholarly journals An Optimized High-Throughput Immuno-Plaque Assay for SARS-CoV-2

2021 ◽  
Vol 12 ◽  
Author(s):  
Alberto A. Amarilla ◽  
Naphak Modhiran ◽  
Yin Xiang Setoh ◽  
Nias Y. G. Peng ◽  
Julian D. J. Sng ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
High Throughput ◽  
Plaque Assay ◽  
Neutralization Assay ◽  
Great Flexibility ◽  
Specific Antibodies ◽  
Research Outcomes ◽  
Global Spread ◽  
Rapid Emergence ◽  
Detection And Quantification

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been identified as the causative agent of coronavirus disease 2019 and is capable of human-to-human transmission and rapid global spread. The rapid emergence and global spread of SARS-CoV-2 has encouraged the establishment of a rapid, sensitive, and reliable viral detection and quantification methodology. Here, we present an alternative assay, termed immuno-plaque assay (iPA), which utilizes a combination of plaque assay and immunofluorescence techniques. We have extensively optimized the conditions for SARS-CoV-2 infection and demonstrated the great flexibility of iPA detection using several antibodies and dual-probing with two distinct epitope-specific antibodies. In addition, we showed that iPA could be utilized for ultra-high-throughput viral titration and neutralization assay within 24 h and is amenable to a 384-well format. These advantages will significantly accelerate SARS-CoV-2 research outcomes during this pandemic period.

scholarly journals Seroprevalence of Hepatitis E Virus in Forestry Workers from Trentino-Alto Adige Region (Northern Italy)

Pathogens ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 568
Author(s):  
Marina Monini ◽  
Fabio Ostanello ◽  
Alessandra Dominicis ◽  
Valentina Tagliapietra ◽  
Gabriele Vaccari ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Northern Italy ◽  
Cumulative Exposure ◽  
Control Group ◽  
Igg Antibodies ◽  
Elisa Kit ◽  
Forestry Workers ◽  
Increased Risk ◽  
Older Subjects

People with some occupational or recreational activities, such as hunters and veterinarians, may have increased risk to be infected by the hepatitis E virus (HEV). The aim of the present study was to establish whether forestry workers could be considered at a higher risk of HEV infection than a control group. One hundred and fifty sera from forestry workers and a control group of 85 sera were analysed by anti-HEV IgG antibodies detection using a commercial ELISA kit. The anti-HEV IgG seroprevalence was 14% for forestry workers and 9.4% for the control group. Comparing the risk of HEV infection in the two groups, there was no difference in the odds ratio. However, the seroprevalence in older subjects was higher in the forestry workers than in the control group. Two sera from forestry workers were also positive for anti-HEV IgM, and, in one of them, HEV-RNA was detected. Our findings showed an increase of seroprevalence with age, which is likely to reflect cumulative exposure to HEV over time. The occupation of forestry workers did not seem to be associated with a higher risk of HEV infection. The study provided new insights into the risk of acquiring HEV in occupational exposure workers with open-air activities.

Prevalence of hepatitis E virus in slaughter-age pigs in Scotland

2014 ◽  
Vol 143 (10) ◽  
pp. 2237-2240 ◽  
Author(s):  
C. CROSSAN ◽  
S. GRIERSON ◽  
J. THOMSON ◽  
A. WARD ◽  
J. NUNEZ-GARCIA ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Partial Sequence ◽  
Genotype 3 ◽  
Serum Samples ◽  
Specific Antibodies ◽  
First Report ◽  
Slaughter Age ◽  
The Uk

SUMMARYThe prevalence of anti-HEV isotype-specific antibodies and viraemia were investigated in serum samples collected from slaughter-age pigs (aged 22–24 weeks) from 23 farms in Scotland. Of 176 serum samples tested, 29·0% (n = 51) were anti-HEV IgG positive, 36·9% (n = 65) anti-HEV IgA positive and 29·0% (n = 51) anti-HEV IgM positive. Overall seroprevalence (anti-HEV IgG+ and/or IgA+ and/or IgM+) was 61·4% (n = 108). HEV RNA was detected in 72/162 serum samples (44·4%). Partial sequence of ORF2 (98 nt) was obtained from eight HEV RNA-positive samples and phylogenetic analysis confirmed that they were all of genotype 3. This is the first report on the prevalence of HEV in pigs in Scotland. Given the increasing incidence of locally acquired HEV infection in the UK, evidence that HEV is a foodborne zoonosis emphasizes the need for surveillance in pigs.

scholarly journals Identification of a novel variant of hepatitis E virus in Austria: sequence, phylogenetic and serological analysis

2000 ◽  
Vol 81 (12) ◽  
pp. 2885-2890 ◽  
Author(s):  
Harald C. Worm ◽  
George G. Schlauder ◽  
Herbert Wurzer ◽  
Isa K. Mushahwar
Keyword(s):  
Recombinant Proteins ◽  
Hepatitis E Virus ◽  
Clinical Presentation ◽  
Acute Viral Hepatitis ◽  
Late Phase ◽  
Hepatitis E ◽  
The Novel ◽  
Serum Samples ◽  
Novel Variant ◽  
Acute Phase Serum

We isolated a novel hepatitis E virus (HEV-Au1) variant from a patient in Austria suffering from acute viral hepatitis, who had no known risk factors for acquiring hepatitis E. The clinical presentation and initial serological findings have been reported previously. In this paper we report the results of sequence and phylogenetic analysis of HEV products from viral RNA isolated from acute phase serum. The results show that HEV-Au1 is significantly divergent from other HEV isolates. The nucleotide identity of analysed fragments from the novel isolate ranges from 76·6 to 78·4% when compared to isolates from endemic regions and 84·6 to 87·9% when compared to isolates from non-endemic regions. Divergent results were obtained when serum samples taken from the convalescent phase of disease were tested with three different immunoassays (EIAs). An EIA based on United States isolate-specific peptides showed enhanced reactivity whereas EIAs based on recombinant proteins derived from prototype HEV strains from Burma and Mexico were unable to detect antibodies to HEV (anti-HEV) in late phase serum. The findings verify the presence of an additional HEV variant in an industrialized country and provide information about possible problems in detecting anti-HEV.

scholarly journals A high-throughput neutralizing assay for antibodies and sera against hepatitis E virus

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Wei Cai ◽  
Zi-Min Tang ◽  
Gui-Ping Wen ◽  
Si-Ling Wang ◽  
Wen-Fang Ji ◽  
...  
Keyword(s):  
High Throughput ◽  
Hepatitis E Virus ◽  
Hepatitis E

Hepatitis E virus (HEV): The novel agent responsible for enterically transmitted non-A, non-B hepatitis

1991 ◽  
Vol 26 (S3) ◽  
pp. 142-147 ◽  
Author(s):  
Gregory R. Reyes ◽  
Patrice O. Yarbough ◽  
Albert W. Tam ◽  
Michael A. Purdy ◽  
Chiao-Chain Huang ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
The Novel ◽  
Novel Agent
Sign in / Sign up

Export Citation Format

Share Document