{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:49:34Z","timestamp":1785408574386,"version":"3.56.0"},"reference-count":35,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T00:00:00Z","timestamp":1780444800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100015811","name":"Science and Technology Department of the Ningxia Hui Autonomous Region","doi-asserted-by":"publisher","award":["2018BBF33001"],"award-info":[{"award-number":["2018BBF33001"]}],"id":[{"id":"10.13039\/100015811","id-type":"DOI","asserted-by":"publisher"}]},{"award":["2018BBF33001"],"award-info":[{"award-number":["2018BBF33001"]}],"id":[{"id":"https:\/\/ror.org\/040ebrp24","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Brucellosis is a globally prevalent zoonotic disease that imposes considerable economic burdens on the livestock industry and remains a significant threat to public health. Although lipopolysaccharide (LPS)-based serological assays are widely used in routine diagnosis, their inherent limitations\u2014particularly cross-reactivity with other Gram-negative bacteria\u2014underscore the need for the development of diagnostic approaches based on non-LPS antigens. In this study, we developed a trivalent nanobody\u2013horseradish peroxidase (3Nbs-HRP) fusion protein targeting Brucella OMP16 and established a cELISA for the serological detection of bovine brucellosis. The diagnostic performance of the assay was assessed using 204 Brucella antibody-negative and 123 Brucella antibody-positive bovine serum samples. ROC curve analysis yielded a sensitivity of 87.7% and a specificity of 89.4%, with no significant cross-reactivity observed. By employing a recombinant antigen and a 3Nbs-HRP probe, this assay enhances biosafety and demonstrates strong potential for standardization and large-scale application, serving as a complementary tool to conventional LPS-based assays for the surveillance, diagnosis, and control of bovine brucellosis.<\/jats:p>","DOI":"10.3390\/ani16111707","type":"journal-article","created":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T07:58:38Z","timestamp":1780473518000},"page":"1707","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and Preliminary Evaluation of an OMP16-Targeting Trivalent Nanobody-HRP-Based cELISA for Serological Detection of Bovine Brucellosis"],"prefix":"10.3390","volume":"16","author":[{"given":"Gaowa","family":"Wudong","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qing","family":"Lu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunyi","family":"Zhai","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ye","family":"Yuan","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaofang","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanhao","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaping","family":"Jin","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"},{"name":"Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong","family":"Zhou","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"},{"name":"Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aihua","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China"},{"name":"Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Xianyang 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Jiao, H., Zhou, G., Wu, S., Meng, C., Wang, L., Fan, C., Li, J., and Chu, Y. 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