{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T11:02:51Z","timestamp":1784545371762,"version":"3.55.0"},"reference-count":29,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100015397","name":"Inner Mongolia Autonomous Region of China","doi-asserted-by":"publisher","award":["2021ZD0013"],"award-info":[{"award-number":["2021ZD0013"]}],"id":[{"id":"10.13039\/501100015397","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Veterinary Sciences"],"abstract":"<jats:p>To develop an indirect ELISA kit for detecting serum antibodies against bovine herpesvirus 1 (BoHV-1), the gD protein of BoHV-1 was expressed in CHO-S cells and purified for use as the coating antigen. The optimized conditions were as follows: a coating antigen concentration of 2 \u00b5g\/mL; serum dilution at 1:40 with incubation at 37 \u00b0C for 60 min; enzyme-labeled antibody dilution at 1:5000 with incubation at 37 \u00b0C for 60 min; and color development with TMB substrate at 37 \u00b0C for 15 min. The absorbance was then measured with a microplate reader. The kit could detect BoHV-1-positive serum at a dilution of up to 1:3200, showing favorable analytical detection capacity. No cross-reactivity was observed with positive sera against bovine viral diarrhea virus (BVDV), Akabane virus (AKAV), bovine parainfluenza virus type 3 (BPIV3), or lumpy skin disease virus (LSDV), reflecting ideal analytical specificity. Based on 126 field serum samples tested against a commercial IDEXX gB-based ELISA kit, we further calculated the diagnostic sensitivity (83.54%; 95% CI: 73.22\u201390.17%) and diagnostic specificity (100%; 95% CI: 92.43\u2013100%) to quantitatively characterize its clinical detection performance. The ROC curve analysis based on IDEXX gB serostatus rather than true VNT-confirmed infection generated an AUC of 0.912 (95% CI: 0.856\u20130.951), which only reflects inter-assay concordance instead of absolute diagnostic validity. This indirect ELISA can be applied to epidemiological investigation and evaluation of immunization efficacy.<\/jats:p>","DOI":"10.3390\/vetsci13070699","type":"journal-article","created":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T09:32:54Z","timestamp":1784539974000},"page":"699","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment of an Indirect ELISA for Bovine Infectious Rhinotracheitis Based on Eukaryotic Expression of gD Protein in CHO Cells"],"prefix":"10.3390","volume":"13","author":[{"given":"Anyan","family":"Li","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China"},{"name":"Organic Milk Production Base, Inner Mongolia Shengmu Hi-Tech Animal Husbandry Co., Ltd., Hohhot 010111, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shimei","family":"Xu","sequence":"additional","affiliation":[{"name":"Organic Milk Production Base, Inner Mongolia Shengmu Hi-Tech Animal Husbandry Co., Ltd., Hohhot 010111, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting","family":"Li","sequence":"additional","affiliation":[{"name":"Organic Milk Production Base, Inner Mongolia Shengmu Hi-Tech Animal Husbandry Co., Ltd., Hohhot 010111, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1826-6838","authenticated-orcid":false,"given":"Qiaoying","family":"Zeng","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock (Co-Established by Province and Ministry), College of Life Sciences, Inner Mongolia University, Hohhot 010070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Zhang, J., Cao, J., Cao, L., Zhang, C., Gong, L., Kang, X., Li, D., and Zhang, Y. 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