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Pseudorabies (PR) outbreaks have devastated many swine farms in several parts of China since late 2011. The outbreak-associated pseudorabies virus (PRV) variant strains exhibited some typical amino acid changes in glycoprotein E (gE), a diagnostic antigen used for discriminating between PRV-infected and vaccinated animals (DIVA). To counteract the potential impact of epitope variations on current serological diagnostics of PRV, we produced monoclonal antibodies (mAbs) against gE protein of one representative PRV variant strain and developed a blocking immunoperoxidase monolayer assay (b-IPMA) for DIVA. The b-IPMA was based on the inhibition of binding between PRV-infected cells and mAb by PRV-specific antibodies present in clinical swine sera and was validated by comparison with a commercial PRV gpI Antibody Test Kit (IDEXX Laboratories, USA). The diagnostic sensitivity, diagnostic specificity and agreement were determined to be 99.25%, 98.18% and 99.02% respectively upon testing 509 serum samples. b-IPMA detected only PRV-specific antibodies and showed no cross- -reactivity with antibodies elicited by gE-deleted vaccine or other common swine pathogens. Thus, b-IPMA has the potential to be used for high-throughput screening of PRV-infected animals in veterinary clinics.

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Authors and Affiliations

Y.B. Wang

Y.H. Li

Q.M. Li

W.T. Xie

C.L. Guo

J.Q. Guo

R.G. Deng

G.P. Zhang

Porcine insulin receptor substrate 2: molecular cloning, tissues distribution, and functions in hepatocyte and aortic endothelial cells

Z. Yin M. Cai X. Weng Z. Liu G. Zhang

Polish Journal of Veterinary Sciences | 2019 | vol.22 | No 3 | 589–598 | DOI: 10.24425/pjvs.2019.129968

Keywords IRS-2 molecular cloning hepatocytes aortic endothelial cells

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Abstract

Insulin receptor substrate 2 (IRS-2) modulates the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, which controls the suppression of gluconeogenic genes; IRS-2 is also a critical node of insulin signaling. Because of the high homology between pig and human IRS-2, we investigated the expression pattern and function of porcine IRS-2. QPCR and immunoblotting were used to detect the IRS-2 expression level in different tissues. There were high IRS-2 levels in the cerebral cortex, hypothalamus, and cerebellum in the central nervous system. In peripheral tissues, IRS-2 was expressed at relatively high levels in the liver. Immunohistochemistry analysis revealed that IRS-2 was mainly distributed in the hypothalamus and cerebral cortex. Furthermore, IRS-2 knockdown porcine hepatocytes and porcine aortic endothelial cells (PAECs) were generated. The IRS-2 knockdown induced abnormal expression of genes involved in glycolipid metabolism in hepatocytes and reduced the antiatherosclerosis ability in PAECs. In addition, we disrupted IRS-2 in porcine embryonic fibroblasts (PEFs) using the CRISPR/Cas9 genome editing system, before finally generating IRS-2 knockout embryos by somatic cell nuclear transfer (SCNT). Taken together, our results indicate that IRS-2 might be a valuable target to establish diabetes and vascular disease models in the pig.

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Authors and Affiliations

Z. Yin

M. Cai

X. Weng

Z. Liu

G. Zhang

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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Bibliography

Arsawiset S, Teepoo S (2020) Ready-to-use, functionalized paper test strip used with a smartphone for the simultaneous on-site detection of free chlorine, hydrogen sulfide and formaldehyde in wastewater. Anal Chim Acta 1118: 63-72.
Chae C (2004) Postweaning multisystemic wasting syndrome: a review of aetiology, diagnosis and pathology. Vet J 168: 41-49.
Griol A, Peransi S, Rodrigo M, Hurtado J, Bellieres L, Ivanova T, Zurita D, Sánchez C, Recuero S, Hernández A, Simón S, Balka G, Bossis I, Capo A, Camarca A, D’Auria S, Varriale A, Giusti A (2019) Design and development of photonic biosensors for swine viral diseases detection. Sensors (Basel) 19: 3985.
Huang L, Tian S, Zhao W, Liu K, Ma X, Guo J (2020) Multiplexed detection of biomarkers in lateral-flow immunoassays. Analyst 145: 2828-2840.
Huang L, Wei Y, Xia D, Liu D, Zhu H, Wu H, Feng L, Liu C (2019) A broad spectrum monoclonal antibody against porcine circovirus type 2 for antigen and antibody detection. Appl Microbiol Biotechnol 103: 3453-3464.
Jin Q, Yang J, Lu Q, Guo J, Deng R, Wang Y, Wang S, Wang S, Chen W, Zhi Y, Wang L, Yang S, Zhang G (2012) Development of an immunochromatographic strip for the detection of antibodies against porcine circovirus-2. J Vet Diagn Invest 24: 1151-1157.
Johnson J, Higgins A, Navarro A, Huang Y, Esper FL, Barton N, Esch D, Shaw C, Olivo PD, Miao LY (2011) Subtyping influenza A virus with monoclonal antibodies and an indirect immunofluorescence assay. J Clin Microbiol 50: 396-400.
Kaushik A, Tiwari S, Dev Jayant R, Marty A, Nair M (2016) Towards detection and diagnosis of Ebola virus disease at point-of-care. Biosens Bioelectron 75: 254-272.
Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256: 495-497.
Li G, Wang A, Chen Y, Sun Y, Du Y, Wang X, Ding P, Jia R, Wang Y, Zhang G (2021) Development of a colloidal gold-based immunochromatographic strip for rapid detection of severe acute respiratory syndrome coronavirus 2 spike protein. Front Immunol 12: 635677.
Li H, Yang J, Bao D, Hou J, Zhi Y, Yang Y, Ji P, Zhou E, Qiao S, Zhang G (2017) Development of an immunochromatographic strip for detection of antibodies against porcine reproductive and respiratory syndrome virus. J Vet Sci 18: 307-316.
Liu C, Feng H, Liu Y, Chen Y, Yang S, Deng R, Zhang G (2020) Soluble FMDV VP1 proteins fused with calreticulin expressed in Escherichia coli under the assist of trigger factor16 (Tf16) formed into high immunogenic polymers. Int J Biol Macromol 155: 1532-1540.
Liu G, Qiao X, Chang C, Hua T, Wang J, Tang B, Zhang D (2020) Reduction of postweaning multisystemic wasting syndrome-associated clinical symptoms by virus-like particle vaccine against porcine parvovirus and porcine circovirus type 2. Viral Immunol 33: 444-456.
Liu X, Yang J, Li Q, Wang Y, Wang Y, Li G, Shi J, Ding P, Guo J, Deng R, Zhang G (2020) A strip test for the optical determination of influenza virus H3 subtype using gold nanoparticle coated polystyrene latex microspheres. Mikrochim Acta 187: 306.
Ma Y, Ni C, Dzakah EE, Wang HY, Kang K, Tang SX, Wang JH, Wang JF (2016) Development of monoclonal antibodies against HIV-1 p24 protein and its application in colloidal gold immunochromatographic assay for HIV-1 detection. Biomed Res Int 2016: 6743904.
Meyers C, Mo XB, Li XD, Yin B, Deng JH, Tian KG, Yuan A (2019) Structural roles of PCV2 capsid protein N-terminus in PCV2 particle assembly and identification of PCV2 type-specific neutralizing epitope. PLoS Pathog 15: e1007562.
Montagnese C, Barattini P, Giusti A, Balka G, Bruno U, Bossis I, Gelasakis A, Bonasso M, Philmis P, Dénes L, Peransi S, Rodrigo M, Simón S, Griol A, Wozniakowski G, Podgorska K, Pugliese C, Nannucci L, D’Auria S, Varriale A (2019) A diagnostic device for in-situ detection of swine viral diseases: the SWINOSTICS project. Sensors (Basel) 19: 407.
Na G, Hu X, Yang J, Sun Y, Kwee S, Tang L, Xing G, Xing Y, Zhang G (2020) Colloidal gold-based immunochromatographic strip assay for the rapid detection of bacitracin zinc in milk. Food Chem 327: 126879.
Nannucci L, Barattini P, Bossis I, Woźniakowski G, Balka G, Pugliese C (2020) Point-of-service diagnostic technology for detection of swine viral diseases. J Vet Res 64: 15-23.
Shang SB, Jin YL, Jiang XT, Zhou JY, Zhang X, Xing G, He JL, Yan Y (2009) Fine mapping of antigenic epitopes on capsid proteins of porcine circovirus, and antigenic phenotype of porcine circovirus type 2. Mol Immunol 46: 327-334.
Wang Y, Guo J, Qiao S, Li Q, Yang J, Jin Q, Zhang G (2016) GP5 protein-based ELISA for the detection of PRRSV antibodies. Pol J Vet Sci 19: 495-501.
Warren AD, Kwong GA, Wood DK, Lin KY, Bhatia SN (2014) Point-of-care diagnostics for noncommunicable diseases using synthetic urinary biomarkers and paper microfluidics. Proc Natl Acad Sci U S A 111: 3671-3676.
Yang X, Yang J, Wang Y, Li L, Sun Z, Yue Z, Tian F, He L, Hu X (2018) A lateral flow immunochromatographic strip test for rapid detection of oseltamivir phosphate in egg and chicken meat. Sci Rep 8: 16680.
Zhang G, Guo J, Wang X (2009) Immunochromatographic lateral flow strip tests. Methods Mol Biol 504: 169-183.
Zhang G, Wang X, Yang J, Yang Y, Xing G, Li Q, Zhao D, Chai S, Guo J (2006) Development of an immunochromatographic lateral flow test strip for detection of beta-adrenergic agonist Clenbuterol residues. J Immunol Methods 312: 27-33.

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Authors and Affiliations

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

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

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PHASE TRANSITION AND PLASTICITY ENHANCEMENT OF Ti-Cu-BASED BULK METALLIC GLASSES

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GP5 Protein-based ELISA for the Detection of PRRSV Antibodies

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Development of a blocking immunoperoxidase monolayer assay for differentiation between pseudorabies virus-infected and vaccinated animals

Abstract

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Porcine insulin receptor substrate 2: molecular cloning, tissues distribution, and functions in hepatocyte and aortic endothelial cells

Abstract

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

Abstract

Bibliography

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