Development and application of a double-antigen Sandwich ELISA using capsid protein to detect multispecies antibodies against porcine circovirus type 3

doi:10.1016/j.jia.2025.03.016

Journal of Integrative Agriculture

2025, Vol. 24

Issue (10): 4105-4109 DOI: 10.1016/j.jia.2025.03.016

Letter

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Development and application of a double-antigen Sandwich ELISA using capsid protein to detect multispecies antibodies against porcine circovirus type 3

Dexin Li^1*, Junhua Deng^2*, Yufang Li², Liying Hao², Wenqiang Pang³, Zhendong Zhang¹, Yi Yang¹, Xiangdong Li^{1, 2#},Kegong Tian^{2, 3#}

¹Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China

²Luoyang Putai Biotechnology Co., Ltd., Luoyang 471003, China

³National Research Center for Veterinary Medicine, Luoyang 471003, China

Highlights
● A novel double antigen sandwich ELISA (DAgS-ELISA) using baculovirus-expressed PCV3 Cap protein was established, enabling sensitive detection of PCV3 antibodies across diverse animal species.
● A serosurvey in Chinese pigs (2022–2024) revealed a 55.7% overall PCV3 seropositivity rate, significantly higher in sows (86.3%).
● PCV3 antibodies were detected in cattle (7.7%) and dogs (4.4%), and reported for the first time in horses, thereby significantly expanding the known host range of PCV3.

Abstract
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摘要

猪圆环病毒3型（Porcine circovirus type 3, PCV3）是一种新发病原体，可感染猪、犬、牛、小鼠等多种动物。此前，已有多种间接酶联免疫吸附试验（iELISA）用于检测猪体内PCV3抗体。本研究首次以杆状病毒表达系统（BEVS）制备的PCV3衣壳蛋白（Cap）作为包被抗原，建立了一种双抗原夹心ELISA（DAgS-ELISA），用于检测不同动物血清中的PCV3抗体。利用该方法，对2022年至2024年中国17个省份的猪血清样本进行了大规模血清学调查，结果显示猪群PCV3血清阳性率为55.7%，其中母猪阳性率显著高于其他群体，达86.3%。此外，该方法成功检测到牛和犬血清中的PCV3抗体，阳性率分别为7.7%和4.4%。进一步研究还将其应用于马、昆明小鼠及20种野生动物血清样本的检测，首次在马体内检出PCV3抗体，表明PCV3的宿主范围进一步扩大。本研究结果证实了PCV3具有广泛的宿主谱，同时表明DAgS-ELISA是一种高效、可靠的血清学检测工具，对PCV3的流行病学监测及防控具有重要意义。

Received: 30 September 2024 Online: 22 March 2025 Accepted: 26 February 2025

Fund:

This study was supported by the National Key Research and Development Program of China (2023YFD1800500), the Jiangsu Innovative and Entrepreneurial Talent Team Project, China (JSSCTD202224), the 111 Project D18007, and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), China.

About author: Dexin Li, E-mail: lidexin0123@163.com; Junhua Deng, E-mail: deng.junhua@sslab.com.cn; #Correspondence Xiangdong Li, E-mail: 007352@yzu.edu.cn; Kegong Tian, E-mail: tiankg@263.net * These authors contributed equally to this study.

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Cite this article:

Dexin Li, Junhua Deng, Yufang Li, Liying Hao, Wenqiang Pang, Zhendong Zhang, Yi Yang, Xiangdong Li, Kegong Tian. 2025. Development and application of a double-antigen Sandwich ELISA using capsid protein to detect multispecies antibodies against porcine circovirus type 3. Journal of Integrative Agriculture, 24(10): 4105-4109.

Cao X, Huang M, Wang Y, Chen Y, Yang H, Quan F. 2023. Immunogenicity analysis of PCV3 recombinant capsid protein virus-like particles and their application in antibodies detection. International Journal of Molecular Sciences, 24, 10377.

Deng J, Li X, Zheng D, Wang Y, Chen L, Song H, Wang T, Huang Y, Pang W, Tian K. 2018. Establishment and application of an indirect ELISA for porcine circovirus 3. Archives of Virology, 163, 479–482.

Ge M, Ren J, Xie Y L, Zhao D, Fan F C, Song X Q, Li M X, Xiao C T. 2021. Prevalence and genetic analysis of porcine circovirus 3 in China from 2019 to 2020. Frontiers in Veterinary Science, 8, 773912.

Geng S, Luo H, Liu Y, Chen C, Xu W, Chen Y, Li X, Fang W. 2019. Prevalence of porcine circovirus type 3 in pigs in the southeastern Chinese province of Zhejiang. BMC Veterinary Research, 15, 244.

Palinski R, Pineyro P, Shang P, Yuan F, Guo R, Fang Y, Byers E, Hause B M. 2017. A novel porcine circovirus distantly related to known circoviruses is associated with porcine dermatitis and nephropathy syndrome and reproductive failure. Journal of Virology, 91, e01879–16.

Tan C Y, Lin C N, Ooi P T. 2021. What do we know about porcine circovirus 3 (PCV3) diagnosis so far?: A review. Transboundary and Emerging Diseases, 68, 2915–2935.

Zhang S, Wang D, Jiang Y, Li Z, Zou Y, Li M, Yu H, Huang K, Yang Y, Wang N. 2019. Development and application of a baculovirus-expressed capsid protein-based indirect ELISA for detection of porcine circovirus 3 IgG antibodies. BMC Veterinary Research, 15, 79.

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