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Goose astrovirus (GoAstV) is a novel avastrovirus that typically causes gosling gout and results in 2 to 20% mortality. GoAstV capsid protein is the sole structural protein, which is responsible for viral attachment, assembly, maturation as well as eliciting host antibodies. However, the epitopes within capsid protein have not been well studied. In this study, a monoclonal antibody, named 1D7, was generated against GoAstV capsid protein by hybridoma technology. Western blot results showed that this MAb could react with recombinant capsid protein expressed in E. coli. Also, it recognized the precursor of capsid protein, VP90 and VP70, in GoAstV-infected cells. Besides, excellent specificity of MAb 1D7 was further demonstrated in indirect immunofluorescence assay and immunohistochemical analysis. Epitope mapping results revealed that MAb 1D7 recognized the epitope ³³QKVY ³⁶ within Cap protein. Sequence alignment indicated that ³³QKVY ³⁶ is a conserved epitope among the isolates of goose astrovirus type 2 (GoAstV-2), suggesting the potential for its use in GoAstV-2 specific diagnostic assay. These findings may provide some insight into a function of the GoAstV capsid protein and further contribute to the development of diagnostic methods for GoAstV infection.

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Authors and Affiliations

G. Dai

1 2 3

X. Huang

1 3

Q. Liu

1 3

Y. Li

1 3

L. Zhang

1 3

K. Han

1 3

J. Yang

1 3

Y. Liu

1 3

F. Xue

2

D. Zhao

1 2 4 3

Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, No. 50 Zhongling Street, Nanjing City, Jiangsu Province, 210014, PR China
College of Veterinary Medicine, Nanjing Agricultural University, No. 1 Tongwei Road, Nanjing City, Jiangsu Province 210095, PR China
Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, Nanjing City, Jiangsu Province, 210014, PR China
Institute of Life Sciences, Jiangsu University, No. 301 Xuefu Road, Zhenjiang, Jiangsu Province, 212013, PR China

Generation and characterization of a monoclonal antibody against duck Tembusu virus envelope protein

T. Xu Y. Liu K. Han D. Zhao J. Yang Y. Li Q. Liu X. Huang K. Bi

Polish Journal of Veterinary Sciences | 2016 | No 4 | DOI: 10.1515/pjvs-2016-0109

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Authors and Affiliations

T. Xu

Y. Liu

K. Han

D. Zhao

J. Yang

Y. Li

Q. Liu

X. Huang

K. Bi

Development of an indirect enzyme-linked immunosorbent assay for the detection of novel chicken orthoreovirus

H. Liu Z. Wei J. Yang Y. Wang J. Hu Y. Tang Y. Diao

Polish Journal of Veterinary Sciences | 2022 | vol. 25 | No 1 | 109-118 | DOI: 10.24425/pjvs.2022.140847

Keywords novel avian orthoreovirus σC gene recombinant plasmid indirect enzyme linked immunosorbent assay epidemiological investigation

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Abstract

A novel avian orthoreovirus (N-ARV) variant characterized with obvious arthritis and synovial inflammation, was isolated from Shandong, China in May 2016. It caused chicken poor growth and enormous economic losses to the poultry industry of China. However, there are few effective methods for detecting the antibody levels of N-ARV. In this study, a viral structural protein σC was expressed using the prokaryotic expression vector pET32a (+). The target protein was obtained by inducing for 6 hours at an IPTG concentration of 0.6mM. The optimal dilution of the coating antigen and serum antibody were determined to be 1000 fold and 10 fold respectively. A specificity test showed that there was no positive reactivity between N-ARV and other pathogens, and when the positive serum was diluted 100 times detection results were still checkable. The repeatability of this method was determined by the inter assay and intra assay tests with variability ranging from 4.85% to 7.93%. In conclusion, this indirect enzyme linked immunosorbent assay (ELISA) will be useful for large-scale serological surveys and monitoring antibody levels in N-ARV infection.

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Authors and Affiliations

H. Liu

1 2 3

e-mail:

ORCID:

Z. Wei

1 2 3

J. Yang

1 2 3

Y. Wang

1 2 3

e-mail:

ORCID:

J. Hu

1 2 3

Y. Tang

1 2 3

Y. Diao

1 2 3

College of Veterinary Medicine, Shandong Agricultural University, No.61 Daizong Street, Tai’an 271018, China
Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University, No.61 Daizong Street, Tai’an 271018, China
Shandong Provincial Engineering Technology Research Center of Animal Disease Control and Prevention, Shandong Agricultural University, No.61 Daizong Street, Tai’an 271018, China

Rapid screening of monoclonal antibodies against porcine circovirus type 2 using colloidal gold-based paper test

Q.Y. Jin L.L. Feng Y.B. Wang P. Li J.F. Yang M. Teng S.J. Chai G.X. Xing G.P. Zhang

Polish Journal of Veterinary Sciences | 2022 | vol. 25 | No 1 | 27-34 | DOI: 10.24425/pjvs.2022.140837

Keywords porcine circovirus type 2 (PCV2) screening of MAbs paper test

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Abstract

A proof of concept for using paper test as a suitable method in the production of monoclonal antibodies (MAbs) is reported. The paper test which detects antibodies against porcine circovirus type 2 (PCV2) using colloidal gold-labelled capsid protein as the antigen probe was applied exclusively in the screening of anti-PCV2 MAbs. It allowed the detection of 118 single cell clones within 30 min using naked eyes. MAbs with specific binding to authentic epitopes on the virus were selected using a blocking strategy in which the antibody was pre-incubated with PCV2 viral sample before applying to the test paper. Five hybridomas secreting MAbs against the capsid protein were obtained, with only three of them capable of binding to PCV2. The results were validated and confirmed using enzyme-linked immunosorbent assay and immunofluorescence assay. The paper test is simple, rapid, and independent on professional technicians and proves to be an excellent approach for the screening of MAbs against specific targets.

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Authors and Affiliations

Q.Y. Jin

1

L.L. Feng

2

Y.B. Wang

3

P. Li

4

J.F. Yang

1

M. Teng

1

S.J. Chai

1

G.X. Xing

1

G.P. Zhang

1

Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, PR China
Institute of Agricultural Economics and Information, Henan Academy of Agricultural Sciences, Zhengzhou 450002, PR China
School of Public Health, Xinxiang Medical University, Xinxiang 453003, PR China
School of Life Sciences and Basic Medicine, Xinxiang University, Xinxiang 453003, PR China

Effect of experimental infections of various Tembusu virus strains isolated from geese, ducks and chickens on ducklings

Y. Li Q. Liu T. Xu X. Huang X. Liu K. Han Y. Liu J. Yang D. Zhao K. Bi W. Sun

Polish Journal of Veterinary Sciences | 2018 | vol. 21 | No 2 | 389–396 | DOI: 10.24425/122613

Keywords goose-derived duck-derived chicken-derived Tembusu virus pathogenicity

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Abstract

In order to compare the pathogenicity of different Tembusu virus (TMUV) strains from geese, ducks and chickens, 56 5-day-old Cherry Valley ducklings which were divided into 7 groups and infected intramuscularly with 7´105 PFU/ml per duck of six challenge virus stocks. The clinical signs, weight gain, mortality, macroscopic and microscopic lesions, virus loads in sera of 1, 3, 5, 7, 11 and 14 dpi and serum antibody titers were examined. The results showed that these viruses could make the young ducks sick, but the clinical signs differed with the different species-original strains. All the experimental groups lose markedly in weight gain compared to the control, but there were no obvious distinctions in weight gains, as well as macroscopic and microscopic lesions of dead ducks between the infected groups. However, the groups of waterfowl-derived strains (from geese and ducks) showed more serious clinical signs and higher relative expressions of virus loads in sera than those from chicken-derived. The mortality of waterfowl groups was 37.5%, and the greatest mortality of chicken groups was 12.5%. The serum antibodies of the geese-species group JS804 appeared earlier and were higher in the titers than others. Taken toghter, the pathogenicity of waterfowl-derived TMUV was more serious than chicken-derived TMUV and JS804 could be chosen as one TMUV vaccine strain to protect from the infection.

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Authors and Affiliations

Y. Li

Q. Liu

T. Xu

X. Huang

X. Liu

K. Han

Y. Liu

J. Yang

D. Zhao

K. Bi

W. Sun

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Iterative sliding mode observer for sensorless control of five-phase permanent magnet synchronous motor

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GP5 Protein-based ELISA for the Detection of PRRSV Antibodies

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Identification of a linear epitope in the capsid protein of goose astrovirus with monoclonal antibody

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Generation and characterization of a monoclonal antibody against duck Tembusu virus envelope protein

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Development of an indirect enzyme-linked immunosorbent assay for the detection of novel chicken orthoreovirus

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Rapid screening of monoclonal antibodies against porcine circovirus type 2 using colloidal gold-based paper test

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Effect of experimental infections of various Tembusu virus strains isolated from geese, ducks and chickens on ducklings

Abstract

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