Development of a blocking ELISA employing a VP1-specific monoclonal antibody for the detection of DHAV3 antibodies

Duck hepatitis A virus (DHAV) causes an acute and severe infectious disease characterized by liver swelling and hemorrhage, predominantly affecting ducklings under three weeks of age. This disease leads to significant economic losses in the duck farming industry. In China, both DHAV1 and DHAV3 are p...

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Main Authors: Cuiping Song, Tianyuan Li, Jing Wang, Peidong Guo, Wenjing Yang, Ning Tang, Yang Qu, Siyu Li, Xusheng Qiu, Lei Tan, Yingjie Sun, Ying Liao, Chan Ding
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Poultry Science
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Online Access:http://www.sciencedirect.com/science/article/pii/S0032579125003190
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author Cuiping Song
Tianyuan Li
Jing Wang
Peidong Guo
Wenjing Yang
Ning Tang
Yang Qu
Siyu Li
Xusheng Qiu
Lei Tan
Yingjie Sun
Ying Liao
Chan Ding
author_facet Cuiping Song
Tianyuan Li
Jing Wang
Peidong Guo
Wenjing Yang
Ning Tang
Yang Qu
Siyu Li
Xusheng Qiu
Lei Tan
Yingjie Sun
Ying Liao
Chan Ding
author_sort Cuiping Song
collection DOAJ
description Duck hepatitis A virus (DHAV) causes an acute and severe infectious disease characterized by liver swelling and hemorrhage, predominantly affecting ducklings under three weeks of age. This disease leads to significant economic losses in the duck farming industry. In China, both DHAV1 and DHAV3 are prevalent, with DHAV3 being more dominant. Among the three structural proteins of DHAV, the VP1 protein is the most critical as it induces neutralizing antibody production, serves as the binding protein for viral adsorption to cell-specific receptors, and determines viral antigenicity. The serum neutralization (SN) test is the ''gold standard'' for evaluating DHAV vaccine-immune serum; however, it is time-consuming and labor-intensive. To address this limitation, we developed a rapid, sensitive, and specific blocking ELISA (bELISA) for detecting DHAV3 antibodies. This assay utilizes DHAV3 virus-like particles (VLPs) as the coating antigen and the VP1-specific monoclonal antibody 4B8 as the blocking antibody. The bELISA demonstrated high sensitivity and specificity, detecting only DHAV3 antibodies without cross-reactivity with DHAV1 or other viral antibodies. The assay's cutoff value was determined to be 38.21 %, with intra- and inter-batch coefficients of variation below 5 %, indicating excellent reproducibility. The bELISA showed a 100 % positive concordance rate and a 93.65 % negative concordance rate with the SN test, resulting in an overall concordance rate of 96 %. In summary, this study presents the development of a high-quality bELISA for the detection of DHAV3 antibodies. This assay is suitable for clinical diagnosis of DHAV3, evaluation of maternal antibody levels, and assessment of vaccine efficacy in ducklings, offering a valuable tool for disease control and prevention in the duck industry.
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spelling doaj-art-24e2104cbd7f4e9eaa2ed9d3b560b8bf2025-08-20T03:24:44ZengElsevierPoultry Science0032-57912025-06-01104610508010.1016/j.psj.2025.105080Development of a blocking ELISA employing a VP1-specific monoclonal antibody for the detection of DHAV3 antibodiesCuiping Song0Tianyuan Li1Jing Wang2Peidong Guo3Wenjing Yang4Ning Tang5Yang Qu6Siyu Li7Xusheng Qiu8Lei Tan9Yingjie Sun10Ying Liao11Chan Ding12Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR China; College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning, Guangxi, ChinaHarbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Haping Road 678, Harbin 150069, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR China; College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning, Guangxi, ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR ChinaShanghai Veterinary Research Institute, the Chinese Academy of Agricultural Science, Ziyue Road 518, Shanghai 200241, PR China; College of Agriculture and Biology, Shanghai Jiao Tong University, Dongchuan Road 800, Shanghai 200240, China; Corresponding author.Duck hepatitis A virus (DHAV) causes an acute and severe infectious disease characterized by liver swelling and hemorrhage, predominantly affecting ducklings under three weeks of age. This disease leads to significant economic losses in the duck farming industry. In China, both DHAV1 and DHAV3 are prevalent, with DHAV3 being more dominant. Among the three structural proteins of DHAV, the VP1 protein is the most critical as it induces neutralizing antibody production, serves as the binding protein for viral adsorption to cell-specific receptors, and determines viral antigenicity. The serum neutralization (SN) test is the ''gold standard'' for evaluating DHAV vaccine-immune serum; however, it is time-consuming and labor-intensive. To address this limitation, we developed a rapid, sensitive, and specific blocking ELISA (bELISA) for detecting DHAV3 antibodies. This assay utilizes DHAV3 virus-like particles (VLPs) as the coating antigen and the VP1-specific monoclonal antibody 4B8 as the blocking antibody. The bELISA demonstrated high sensitivity and specificity, detecting only DHAV3 antibodies without cross-reactivity with DHAV1 or other viral antibodies. The assay's cutoff value was determined to be 38.21 %, with intra- and inter-batch coefficients of variation below 5 %, indicating excellent reproducibility. The bELISA showed a 100 % positive concordance rate and a 93.65 % negative concordance rate with the SN test, resulting in an overall concordance rate of 96 %. In summary, this study presents the development of a high-quality bELISA for the detection of DHAV3 antibodies. This assay is suitable for clinical diagnosis of DHAV3, evaluation of maternal antibody levels, and assessment of vaccine efficacy in ducklings, offering a valuable tool for disease control and prevention in the duck industry.http://www.sciencedirect.com/science/article/pii/S0032579125003190Blocking ELISA;Duck hepatitis A virus type 3;Antibody detection
spellingShingle Cuiping Song
Tianyuan Li
Jing Wang
Peidong Guo
Wenjing Yang
Ning Tang
Yang Qu
Siyu Li
Xusheng Qiu
Lei Tan
Yingjie Sun
Ying Liao
Chan Ding
Development of a blocking ELISA employing a VP1-specific monoclonal antibody for the detection of DHAV3 antibodies
Poultry Science
Blocking ELISA;Duck hepatitis A virus type 3;Antibody detection
title Development of a blocking ELISA employing a VP1-specific monoclonal antibody for the detection of DHAV3 antibodies
title_full Development of a blocking ELISA employing a VP1-specific monoclonal antibody for the detection of DHAV3 antibodies
title_fullStr Development of a blocking ELISA employing a VP1-specific monoclonal antibody for the detection of DHAV3 antibodies
title_full_unstemmed Development of a blocking ELISA employing a VP1-specific monoclonal antibody for the detection of DHAV3 antibodies
title_short Development of a blocking ELISA employing a VP1-specific monoclonal antibody for the detection of DHAV3 antibodies
title_sort development of a blocking elisa employing a vp1 specific monoclonal antibody for the detection of dhav3 antibodies
topic Blocking ELISA;Duck hepatitis A virus type 3;Antibody detection
url http://www.sciencedirect.com/science/article/pii/S0032579125003190
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