Development and application of an indirect ELISA for detection of antibodies against avian hepatitis E virus

Zhao, Qin; Sun, Yani; Zhao, Jinan; Hu, Shoubin; Zhao, Fengyuan; Chen, Fuyong; Clavijo, Alfonso; Xiao, Yihong

doi:10.1016/j.jviromet.2012.08.026

Cited by 37 publications

(35 citation statements)

References 14 publications

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“…Furthermore, seroconversion in subclinically infected chickens was reported between weeks 17 and 19 of life (Sun et al, 2004a). Interestingly, Zhao et al (2013) reported a decline of seropositive birds with age in birds from flocks with an incidence of BLSD. This could not be observed in the actual study as birds stayed negative until 25 weeks of life and positive birds were detected at 45 weeks until slaughtering.…”

Section: Discussionmentioning

confidence: 99%

Subclinical circulation of avian hepatitis E virus within a multiple-age rearing and broiler breeder farm indicates persistence and vertical transmission of the virus

Troxler

Pać²,

Prokofieva

et al. 2014

Avian Pathology

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In a prospective longitudinal study, a broiler breeder flock and its progeny were monitored for the presence of avian hepatitis E virus (HEV) RNA and antibodies. The flock was part of a multiple-age farm where the presence of avian HEV with clinical signs (increased mortality and decreased egg production) was demonstrated in several previous production cycles. Samples were taken twice at the rearing site and several times at the production site from broiler breeders including cockerels and day-old chicks. The samples were investigated by conventional and real-time reverse transcriptase-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) and histological methods. At all time points, samples from the hens were positive for avian HEV RNA. The birds did not show any clinical signs, even though histopathological lesions of non-specific aetiology in the liver and spleen could be demonstrated. A significant increase in the number of positive birds and viral load was seen in week 45, in accordance with an increase in antibody titres. In comparison, cockerels investigated in week 62 tested negative by RT-PCR and ELISA. Avian HEV RNA was also detected in day-old chicks hatched from eggs laid in week 25, indicating vertical transmission. All partial helicase and capsid sequences retrieved within this study clustered together and were identical to previous sequences obtained from the same multiple-age farm. In conclusion, avian HEV persisted on the farm over years and circulated between the rearing and the production sites without causing any clinical signs although high viral loads in the adult hens were observed.

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Section: Discussionmentioning

confidence: 99%

Subclinical circulation of avian hepatitis E virus within a multiple-age rearing and broiler breeder farm indicates persistence and vertical transmission of the virus

Troxler

Pać²,

Prokofieva

et al. 2014

Avian Pathology

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“…Antisera against avian, human, and swine HEVs were separately collected from specific-pathogen-free (SPF) chickens infected experimentally with genotype 3 avian HEV isolated from China (CaHEV) (31), from human beings infected naturally by genotype 4 human HEVs, which were identified by reverse transcription-PCR (RT-PCR) as previously described (32) in the Affiliated Hospital of Shandong Agricultural University (33), and from swine herds infected naturally by genotype 4 swine HEV in Shandong Province (34), respectively. These serum samples were analyzed by indirect enzyme-linked immunosorbent assays (ELISAs) using the C-terminal 268 aa of avian and swine HEV capsid proteins (aORF2-C and sORF2-C, respectively), peptide 1 ( 389 GNQHVD DRPSPAPAPKRALGTL 410 ), or peptide 2 ( 461 SVDWTKATVDGVQVKT VDASSGSNRFAALPAF 492 ) as the coating antigen, as described in previous studies (30,31,34). Peptide 1 and peptide 2 included full-length antigenic domains I (common to avian, swine, and human HEVs) and II (unique to avian HEV), respectively (26,30).…”

Section: Methodsmentioning

confidence: 99%

“…To test whether the six MAbs would cross-react with avian and swine HEV capsid proteins, aORF2-C and sORF2-C were expressed from a bacterial system and purified as previously described (31,34).…”

Section: Methodsmentioning

confidence: 99%

“…To assess the protective capacity of the synthesized peptides containing the common epitopes, an animal immunization study was performed. Seven-week-old SPF chickens (n ϭ 30) were purchased from the Beijing Meiliyaweitong Experimental Animal Technology Company and confirmed to be negative for avian HEV RNA and antibodies by using RT-PCR and indirect ELISA (12,31). Chickens were divided into 5 groups (6/group) and housed at 2 chickens per cage.…”

Section: Methodsmentioning

confidence: 99%

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Characterization of Two Novel Linear B-Cell Epitopes in the Capsid Protein of Avian Hepatitis E Virus (HEV) That Are Common to Avian, Swine, and Human HEVs

Wang

Zhao

et al. 2015

J Virol

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Antisera raised against the avian hepatitis E virus (HEV) capsid protein are cross-reactive with human and swine HEV capsid proteins. In this study, two monoclonal antibodies (MAbs) against the avian HEV capsid protein, namely, 3E8 and 1B5, were shown to cross-react with the swine HEV capsid protein. The motifs involved in binding both MAbs were identified and characterized using phage display biopanning, peptide synthesis, and truncated or mutated protein expression, along with indirect enzyme-linked immunosorbent assay (ELISA) and Western blotting. The results showed that the I/VPHD motif is a necessary core sequence and that P and H are two key amino acids for recognition by MAb 3E8. The VKLYM/TS motif is the minimal amino acid sequence necessary for recognition by MAb 1B5. Cross-reactivity between the two epitopes and antibodies against avian, swine, and human HEVs in sera showed that both epitopes are common to avian, swine, and human HEVs. In addition, amino acid sequence alignment of the capsid proteins revealed that the key motifs of both novel epitopes are the same in HEVs from different animal species, predicting that they may be common to HEV isolates from boars, rabbits, rats, ferrets, mongooses, deer, and camels as well. Protein modeling analysis showed that both epitopes are at least partially exposed on the surface of the HEV capsid protein. Protective capacity analysis demonstrated that the two epitopes are nonprotective against avian HEV infection in chickens. Collectively, these studies characterize two novel linear B-cell epitopes common to avian, swine, and human HEVs, which furthers the understanding of HEV capsid protein antigenic structure. IMPORTANCEMore and more evidence indicates that the host range diversity of hepatitis E virus (HEV) is a global public health concern. A better understanding of the antigenic structure of the HEV capsid protein may improve disease diagnosis and prevention. In this study, binding site mapping and localization as well as the antigenic biology of two novel linear B-cell epitopes common to several different species of HEV were characterized. These findings partially reveal the antigenic structure of the HEV capsid protein and provide potential applications for the development of diagnostics and interventions for HEV infection.H epatitis E is usually an acute and self-limiting disease in humans, is an epidemic in many developing countries in Asia and Africa (1-3), and occurs sporadically in some industrialized countries (4-6). The causative agent, hepatitis E virus (HEV), belongs to the Hepeviridae family and is a single-stranded, positivesense RNA virus. The viral genome contains three open reading frames (ORFs), ORF1, -2, and -3, encoding a viral nonstructural protein, a capsid protein, and a small multifunctional phosphoprotein (2, 7), respectively. The proteins encoded by ORF2 and ORF3 are produced from a bicistronic subgenomic (SG) mRNA, and the coding region of ORF2 overlaps ORF3, but neither overlaps ORF1 (8, 9). To date, complete genomes of m...

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“…All flocks exhibited a reduced rate of egg production and hepatitis syndrome. All serum samples were tested for anti-avian HEV antibodies using a previously developed indirect enzyme-linked immunosorbent assay (iELISA) [20] and OD 450nm values of these serum samples were determined (Additional file 1). Of 1995 serum samples, 700 (35.09%) were positive for anti-avian HEV antibodies (Table 1).…”

Section: Resultsmentioning

confidence: 99%

Seroprevalence of avian hepatitis E virus and avian leucosis virus subgroup J in chicken flocks with hepatitis syndrome, China

Sun

Liu

et al. 2016

BMC Vet Res

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BackgroundFrom 2014 to 2015 in China, many broiler breeder and layer hen flocks exhibited a decrease in egg production and some chickens developed hepatitis syndrome including hepatomegaly, hepatic necrosis and hemorrhage. Avian hepatitis E virus (HEV) and avian leucosis virus subgroup J (ALV-J) both cause decreasing in egg production, hepatomegaly and hepatic hemorrhage in broiler breeder and layer hens. In the study, the seroprevalence of avian HEV and ALV-J in these flocks emerging the disease from Shandong and Shaanxi provinces were investigated.ResultsA total of 1995 serum samples were collected from 14 flocks with hepatitis syndrome in Shandong and Shaanxi provinces, China. Antibodies against avian HEV and ALV-J in these serum samples were detected using iELISAs. The seroprevalence of anti-avian HEV antibodies (35.09%) was significantly higher than that of anti-ALV-J antibodies (2.16%) (p = 0.00). Moreover, the 43 serum samples positive for anti-ALV-J antibodies were all also positive for anti-avian HEV antibodies. In a comparison of both provinces, Shandong chickens exhibited a significantly higher seroprevalence of anti-avian HEV antibodies (42.16%) than Shaanxi chickens (26%) (p = 0.00). In addition, the detection of avian HEV RNA and ALV-J cDNA in the liver samples from the flocks of two provinces also showed the same results of the seroprevalence.ConclusionsIn the present study, the results showed that avian HEV infection is widely prevalent and ALV-J infection is endemic in the flocks with hepatitis syndrome from Shandong and Shaanxi provinces of China. These results suggested that avian HEV infection may be the major cause of increased egg drop and hepatitis syndrome observed during the last 2 years in China. These results should be useful to guide development of prevention and control measures to control the diseases within chicken flocks in China.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-016-0892-4) contains supplementary material, which is available to authorized users.

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Development and application of an indirect ELISA for detection of antibodies against avian hepatitis E virus

Cited by 37 publications

References 14 publications

Subclinical circulation of avian hepatitis E virus within a multiple-age rearing and broiler breeder farm indicates persistence and vertical transmission of the virus

Subclinical circulation of avian hepatitis E virus within a multiple-age rearing and broiler breeder farm indicates persistence and vertical transmission of the virus

Characterization of Two Novel Linear B-Cell Epitopes in the Capsid Protein of Avian Hepatitis E Virus (HEV) That Are Common to Avian, Swine, and Human HEVs

Seroprevalence of avian hepatitis E virus and avian leucosis virus subgroup J in chicken flocks with hepatitis syndrome, China

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