{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,6]],"date-time":"2026-11-06T22:05:54Z","timestamp":1794002754449,"version":"4.8.0"},"reference-count":49,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T00:00:00Z","timestamp":1787529600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Major Science and Technology Special Project of Xinjiang Uygur Autonomous Region","award":["2024A02005"],"award-info":[{"award-number":["2024A02005"]}]},{"name":"Xinjiang Talent Development Fund","award":["ZZYD2023010"],"award-info":[{"award-number":["ZZYD2023010"]}]},{"award":["ZZYD2023010"],"award-info":[{"award-number":["ZZYD2023010"]}],"id":[{"id":"https:\/\/ror.org\/01rkhv978","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Prevention and Control of Emerging and Major Infectious Diseases\u2014National Science and Technology Major Project","award":["2025ZD01900100"],"award-info":[{"award-number":["2025ZD01900100"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Rotavirus A (RVA) causes severe diarrhea in donkey foals, but rapid quantitative antigen assays for vaccine development and manufacturing control are lacking. We developed a VP6-based antigen-capture ELISA (acELISA) for donkey-origin RVA using rabbit anti-RVA polyclonal IgG as the capture antibody and horseradish peroxidase-conjugated mouse anti-VP6 monoclonal antibody 3F6 as the detector antibody. Assay conditions were optimized, and performance was evaluated through standard-curve analysis, specificity and repeatability testing, Western blot (WB) comparison, median tissue culture infectious dose (TCID50), reverse transcription quantitative polymerase chain reaction (RT-qPCR), ultrafiltration membrane screening, and pilot-scale tangential flow filtration (TFF) monitoring. The optimized acELISA detected VP6 at 1 ng\/mL and showed a linear range of 1\u201325 ng\/mL, with no cross-reactivity with eight common equine pathogens, and intra- and inter-assay coefficients of variation below 10%. The acELISA estimates were consistent with Western blot densitometry. In process applications, the assay supported the selection of a 100 kDa membrane and monitored pilot-scale TFF, yielding 75.69% VP6 recovery and an 18-fold increase in antigen purity. This acELISA provides a specific and practical in-process tool for quantitative antigen monitoring in donkey rotavirus vaccine production and is intended to complement, rather than replace, infectivity- and genome-based assays.<\/jats:p>","DOI":"10.3390\/microorganisms14091874","type":"journal-article","created":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T08:03:41Z","timestamp":1787558621000},"page":"1874","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Development and Validation of an Antigen-Capture ELISA for Quantitative Detection of Donkey-Origin Rotavirus: Application in Vaccine Manufacturing Process Monitoring"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6867-6598","authenticated-orcid":false,"given":"Zenan","family":"Zhang","sequence":"first","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9496-2267","authenticated-orcid":false,"given":"Xing","family":"Guo","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6274-1857","authenticated-orcid":false,"given":"Kui","family":"Guo","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1507-737X","authenticated-orcid":false,"given":"Wei","family":"Guo","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"},{"name":"Institute of Western Agriculture, The Chinese Academy of Agricultural Sciences, Changji 831100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1686-6189","authenticated-orcid":false,"given":"Cheng","family":"Du","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3862-2003","authenticated-orcid":false,"given":"Yan","family":"Yang","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-3974-0394","authenticated-orcid":false,"given":"Wenjing","family":"Dong","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6479-6659","authenticated-orcid":false,"given":"Xiaoyu","family":"Chu","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7281-0369","authenticated-orcid":false,"given":"Yuezhi","family":"Lin","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4521-4099","authenticated-orcid":false,"given":"Xiaojun","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 150069, China"},{"name":"Institute of Western Agriculture, The Chinese Academy of Agricultural Sciences, Changji 831100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.vetmic.2013.07.010","article-title":"Equine Rotaviruses\u2014Current Understanding and Continuing Challenges","volume":"167","author":"Bailey","year":"2013","journal-title":"Vet. 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