{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,31]],"date-time":"2026-05-31T08:00:48Z","timestamp":1780214448487,"version":"3.54.0"},"reference-count":47,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,5,31]],"date-time":"2026-05-31T00:00:00Z","timestamp":1780185600000},"content-version":"vor","delay-in-days":150,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32072871"],"award-info":[{"award-number":["32072871"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100019091","name":"Key Research and Development Program of Hunan Province of China","doi-asserted-by":"publisher","award":["2023NK2017"],"award-info":[{"award-number":["2023NK2017"]}],"id":[{"id":"10.13039\/501100019091","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transboundary and Emerging Diseases"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:p>\n                    Porcine circovirus 2 (PCV2) remains a major swine pathogen, and interpretation of capsid protein (Cap)\u2013based serology is challenging in vaccinated populations because vaccine\u2010induced Cap antibodies overlap with replication\u2010associated serological signals. To address this issue, we developed two fully automated magnetic particle\u2013based chemiluminescence immunoassays (CLIAs), including a Cap virus\u2010like particle (VLP)\u2013based assay (VLPs\u2010CLIA) for evaluating Cap antibody response intensity and a replication\u2010associated protein (Rep)\u2013based assay (rRep\u2010CLIA) for assessing replication\u2010associated serological exposure or pressure. We produced self\u2010assembled PCV2 Cap VLPs and recombinant Rep in a prokaryotic system and covalently coupled them to carboxylated magnetic beads for fully automated CLIAs. VLPs\u2010CLIA was calibrated and reported in activity units (U), whereas rRep\u2010CLIA was interpreted as a threshold\u2010based qualitative assay using a relative light unit (RLU) cut\u2010off. Analytical performance and agreement with reference enzyme\u2010linked immunosorbent assays (ELISAs) were evaluated using longitudinal and field serum samples. VLPs\u2010CLIA detected antibodies up to 1:25,600, approximately twofold higher than the reference ELISA (1:12,800), with 95.68% agreement (Kappa\u2009=\u20090.876,\n                    <jats:italic>p<\/jats:italic>\n                    \u2009 &lt; 0.001). rRep\u2010CLIA agreed well with Rep\u2010ELISA (94.86%; Kappa\u2009=\u20090.846,\n                    <jats:italic>p<\/jats:italic>\n                    \u2009 &lt; 0.001) and provided a complementary signal consistent with replication\u2010associated exposure. Both assays showed good precision (coefficients of variation [CVs] ranging from 2.83% to 6.94%) and good reagent stability. In field samples, VLPs\u2010CLIA tracked postvaccination Cap kinetics, whereas Rep seropositivity increased with age and exposure background, suggesting heterogeneous replication\u2013associated pressure among herds. The two assays together support complementary herd\u2010level interpretation of vaccine\u2010induced Cap immunity and replication\u2010associated serological exposure, particularly in Cap\u2010vaccinated populations.\n                  <\/jats:p>","DOI":"10.1155\/tbed\/3733817","type":"journal-article","created":{"date-parts":[[2026,5,31]],"date-time":"2026-05-31T07:19:12Z","timestamp":1780211952000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of Magnetic Particle\u2013Based Chemiluminescence Immunoassays for Detecting PCV2 VLP\u2010 and Rep\u2010Specific Antibodies and Their Combined Use for Immune Evaluation and Replication\u2010Associated Exposure Assessment"],"prefix":"10.1155","volume":"2026","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-6558-1024","authenticated-orcid":false,"given":"Mingxi","family":"Gou","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-4141-5462","authenticated-orcid":false,"given":"Xiaoqing","family":"Song","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-8660-4031","authenticated-orcid":false,"given":"Jie","family":"Fan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-0420-3114","authenticated-orcid":false,"given":"Zhixiong","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-4357-0459","authenticated-orcid":false,"given":"Zihao","family":"Kuang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-6007-331X","authenticated-orcid":false,"given":"Mengyao","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-4448-6151","authenticated-orcid":false,"given":"Zhongxin","family":"Fan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7656-6858","authenticated-orcid":false,"given":"Meng","family":"Ge","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,5,31]]},"reference":[{"key":"e_1_2_11_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2012.12.033"},{"key":"e_1_2_11_2_2","doi-asserted-by":"publisher","DOI":"10.1079\/AHR2005106"},{"key":"e_1_2_11_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tvjl.2004.01.012"},{"key":"e_1_2_11_4_2","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.19536-0"},{"key":"e_1_2_11_5_2","doi-asserted-by":"publisher","DOI":"10.1136\/vr.146.2.40"},{"key":"e_1_2_11_6_2","first-page":"44","article-title":"Isolation 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Canadienne"},{"key":"e_1_2_11_7_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12917-025-04928-0"},{"key":"e_1_2_11_8_2","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines11081308"},{"key":"e_1_2_11_9_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12917-023-03648-7"},{"key":"e_1_2_11_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/bab.1491"},{"key":"e_1_2_11_11_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-016-2816-9"},{"key":"e_1_2_11_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2012.04.022"},{"key":"e_1_2_11_13_2","doi-asserted-by":"publisher","DOI":"10.1007\/s11262-014-1099-z"},{"key":"e_1_2_11_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2010.05.013"},{"key":"e_1_2_11_15_2","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens9121049"},{"key":"e_1_2_11_16_2","doi-asserted-by":"publisher","DOI":"10.3390\/v14092005"},{"key":"e_1_2_11_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.18.9885-9893.2003"},{"key":"e_1_2_11_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2012.09.001"},{"key":"e_1_2_11_19_2","doi-asserted-by":"publisher","DOI":"10.3389\/fchem.2021.813219"},{"key":"e_1_2_11_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.00809-24"},{"key":"e_1_2_11_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2021.114269"},{"key":"e_1_2_11_22_2","first-page":"574","article-title":"Establishment of Competitive Chemiluminescent Enzyme Lmmunoassay for Detecting Antibodies Against gB Protein of Pseudorabies Virus","volume":"51","author":"Zhenyuan M.","year":"2020","journal-title":"Acta Veterinaria et Zootechnica Sinica"},{"key":"e_1_2_11_23_2","first-page":"1834","article-title":"Establishment of Double-Antigen Sandwich Chemiluminescent Enzyme Immunoassay for Antibody Detection Against Classical Swine Fever Virus E\n                     rns\n                   Antigen","volume":"45","author":"Zihan Y.","year":"2025","journal-title":"Chinese Journal of Veterinary Science"},{"key":"e_1_2_11_24_2","first-page":"63","article-title":"An Indirect ELISA Established Using the Capsid Protein of PCV2 for Detection of Antibodies to Porcine Circovirus 2","volume":"39","author":"Meng G.","year":"2013","journal-title":"Journal of Hunan Agricultural University"},{"key":"e_1_2_11_25_2","first-page":"894","article-title":"Development of an Indirect ELISA for Detecting Antibodies Against Porcine Circovirus 2 Based on Recombinant Rep Protein","volume":"34","author":"Jie L.","year":"2012","journal-title":"Chinese Journal of Preventive Veterinary Medicine"},{"key":"e_1_2_11_26_2","article-title":"ICH Guideline M10 on Bioanalytical Method Validation and Study Sample Analysis (Step 5)","volume":"6","author":"ICH Member","year":"2023","journal-title":"European Medicines Agency"},{"key":"e_1_2_11_27_2","volume-title":"In, Manual of Diagnostic Tests and Vaccines for Terrestrial Animals","author":"WOAH","year":"2023"},{"key":"e_1_2_11_28_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-animal-031412-103720"},{"key":"e_1_2_11_29_2","doi-asserted-by":"publisher","DOI":"10.3390\/v11020185"},{"key":"e_1_2_11_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2010.03.025"},{"key":"e_1_2_11_31_2","unstructured":"SiruiT. Development Technology of Porcine Circovirus Type 2 and Development of Diagnostic Kits 2022 Zhejiang Sci-Tech University."},{"key":"e_1_2_11_32_2","doi-asserted-by":"publisher","DOI":"10.3390\/vetsci12010040"},{"key":"e_1_2_11_33_2","doi-asserted-by":"publisher","DOI":"10.1039\/c0an00025f"},{"key":"e_1_2_11_34_2","article-title":"A Chemiluminescence Detection Kit for Porcine Circovirus Type 2","volume":"9","author":"Jianhong S.","year":"2024","journal-title":"BMC Veterinary Research"},{"key":"e_1_2_11_35_2","article-title":"A Homogeneous Competitive Chemiluminescence Immunoassay Kit for Porcine Circovirus Type 2 Antibody Detection, Its Preparation Method and Application","volume":"7","author":"Jianhong S.","year":"2020","journal-title":"Journal of the American Chemical Society"},{"key":"e_1_2_11_36_2","unstructured":"CongL. Establishment and Preliminary Application of a Chemiluminescent Detection Method for Porcine Circovirus Type 2 Antibodies 2024 Zhongkai University of Agriculture and Engineering."},{"key":"e_1_2_11_37_2","article-title":"A Tube-Type Chemiluminescence Immunoassay Kit for Detecting Porcine Circovirus Type 2 Antibodies and Its Method of use","volume":"12","author":"Shengjun L.","year":"2023","journal-title":"BMC Veterinary Research"},{"key":"e_1_2_11_38_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2011.11.001"},{"key":"e_1_2_11_39_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.2001.1203"},{"key":"e_1_2_11_40_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2011.10.012"},{"key":"e_1_2_11_41_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1013244"},{"key":"e_1_2_11_42_2","first-page":"1216","article-title":"Determination of Immunogenicity and Neutralizing Activity of the Rep and Rep\u2019 Proteins Derived From Porcine Circovirus Type 2","volume":"42","author":"Dan L.","year":"2012","journal-title":"Chinese Veterinary Science"},{"key":"e_1_2_11_43_2","first-page":"916","article-title":"Immune Protection of Porcine Circovirus Type 2 Rep\/Rep\u2019 and Cap Protein as a Subunit Vaccines","volume":"35","author":"Dan L.","year":"2013","journal-title":"Chinese Journal of Preventive Veterinary Medicine"},{"key":"e_1_2_11_44_2","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.001230"},{"key":"e_1_2_11_45_2","first-page":"2061","article-title":"Prokaryotic Expression and Protein Activity of Porcine Circovirus Type 3 Rep","volume":"55","author":"Yang H.","year":"2024","journal-title":"Acta Veterinaria et Zootechnica Sinica"},{"key":"e_1_2_11_46_2","first-page":"2107","article-title":"Prokaryotic Expression of Rep Protein of Porcine Circovirus Type 3 and Preparation of Polyclonal Antibodies Against Rep Protein","volume":"39","author":"Kaiqi L.","year":"2019","journal-title":"Chinese Journal of Veterinary Science"},{"key":"e_1_2_11_47_2","doi-asserted-by":"publisher","DOI":"10.1186\/1743-422X-7-166"}],"container-title":["Transboundary and Emerging Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1155\/tbed\/3733817","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1155\/tbed\/3733817","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1155\/tbed\/3733817","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,31]],"date-time":"2026-05-31T07:19:20Z","timestamp":1780211960000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1155\/tbed\/3733817"}},"subtitle":[],"editor":[{"given":"Zhong","family":"Peng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,1]]},"references-count":47,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2026,1]]}},"alternative-id":["10.1155\/tbed\/3733817"],"URL":"https:\/\/doi.org\/10.1155\/tbed\/3733817","archive":["Portico"],"relation":{},"ISSN":["1865-1674","1865-1682"],"issn-type":[{"value":"1865-1674","type":"print"},{"value":"1865-1682","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1]]},"assertion":[{"value":"2026-02-28","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-12","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-31","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"3733817"}}