{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T04:26:15Z","timestamp":1787199975955,"version":"build-2736575974"},"reference-count":42,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T00:00:00Z","timestamp":1787184000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:sec>\n                    <jats:title>Inroduction<\/jats:title>\n                    <jats:p>Infectious diseases caused by bovine viral diarrhea virus (BVDV), Akabane virus (AKAV), bovine norovirus (BNoV), bovine enterovirus (BEV), and bovine coronavirus (BCoV) significantly threaten the cattle industry, resulting in substantial economic losses. These pathogens often present similar clinical signs, such as diarrhea, vomiting, and reproductive disorders in pregnant cattle, and frequent covert or mixed infections further complicate accurate diagnosis. Therefore, rapid, sensitive, and field\u2011deployable diagnostic methods are essential for effective disease surveillance and control in the cattle industry.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>In this study, we report for the first time the establishment of a TaqMan\u2011based real\u2011time quantitative PCR (qPCR) assay that enables simultaneous detection of these five bovine viruses. Multiple sequence alignment of conserved genomic regions was performed, and virus\u2011specific primers and probes were designed and optimized using Beacon Designer 7 software. Subsequently, a TaqMan\u2011based multiplex real\u2011time qPCR assay was established for simultaneous detection of BVDV, AKAV, BNoV, BEV, and BCoV. The established detection method was applied to 200 clinical samples collected from 10 farms in multiple regions of Jilin Province.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The results showed that the detection rates for BVDV, AKAV, BNoV, BEV, and BCoV were 33.50%, 0.50%, 4.50%, 7.50%, and 12.00%, respectively. Mixed infections were detected in 9 samples co\u2011infected with two of the five pathogens, with an overall mixed infection rate of 4.50%. Compared with conventional PCR, coincidence rates were 100% for BVDV, AKAV, BNoV, BEV, and BCoV.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>These findings indicate that the TaqMan multiplex real\u2011time qPCR assay developed here demonstrates favorable specificity, sensitivity, and reproducibility. This assay enables efficient detection and surveillance of bovine viruses, offering a reliable technical tool for the diagnosis and control of corresponding viral diseases in cattle.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fvets.2026.1906498","type":"journal-article","created":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T04:15:34Z","timestamp":1787199334000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and epidemiological investigation of a TaqMan-based multiplex real-time quantitative PCR assay for simultaneous detection of five bovine viruses (BVDV, AKAV, BNoV, BEV, and BCoV)"],"prefix":"10.3389","volume":"13","author":[{"given":"Hanjia","family":"Zhang","sequence":"first","affiliation":[{"name":"College of Life Sciences, Jilin Agricultural University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanbing","family":"Bai","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Jilin Agricultural University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fan","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Jilin Agricultural University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuanyi","family":"Liu","sequence":"additional","affiliation":[{"name":"Sanjiang Laboratory, Jilin Agricultural University","place":["Changchun, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunling","family":"Guo","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Jilin Agricultural University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linyue","family":"Zhou","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Jilin Agricultural University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruichi","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Jilin Agricultural University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sisi","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Jilin Agricultural University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingfeng","family":"Meng","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Jilin Agricultural University","place":["Changchun, Jilin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Dong","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Jilin Agricultural University","place":["Changchun, Jilin, China"]},{"name":"Sanjiang Laboratory, Jilin Agricultural University","place":["Changchun, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,8,20]]},"reference":[{"key":"ref1","article-title":"Epidemiological Investigation and Analysis of Bovine Coronavirus in Jilin Province","author":"Sun","year":"2022"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"1143","DOI":"10.16303\/j.cnki.1005-4545.2022.06.07","article-title":"Epidemiological investigation of mixed infection with bovine viral diarrhea virus and bovine enterovirus in Jilin Province","volume":"42","author":"Dong","year":"2022","journal-title":"Chin J Vet 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