{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T04:31:57Z","timestamp":1786422717699,"version":"build-2736575974"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T00:00:00Z","timestamp":1786406400000},"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>Introduction<\/jats:title>\n                    <jats:p>Bovine brucellosis and bovine tuberculosis are important zoonotic diseases that pose serious threats to human and animal health and cause substantial economic losses in the livestock industry. Therefore, a rapid diagnostic method suitable for the simultaneous field detection of both diseases is urgently needed.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Fluorescent microspheres conjugated with recombinant streptococcal protein G (rSPG) were prepared and used as tracers.\n                      <jats:italic>Brucella<\/jats:italic>\n                      lipopolysaccharide (LPS) and\n                      <jats:italic>Mycobacterium bovis<\/jats:italic>\n                      MPB70 protein were coated onto test lines 1 and 2 (TL1 and TL2), respectively, whereas rSPG was coated onto the control line (CL). This configuration enabled the development of a dual-antibody rapid test strip for the simultaneous detection of\n                      <jats:italic>Brucella<\/jats:italic>\n                      spp. and\n                      <jats:italic>M. bovis<\/jats:italic>\n                      antibodies. The optimal reaction conditions and performance characteristics of the test strip were systematically evaluated.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results and discussion<\/jats:title>\n                    <jats:p>\n                      The dual rapid test strip enabled detection to be completed within 20 min. The optimal coating concentrations were determined to be 0.5 mg\/mL for the Brucella test line (TL1) and 1.0 mg\/mL for the\n                      <jats:italic>M. bovis<\/jats:italic>\n                      test line (TL2), and the optimal conjugation concentration of fluorescent microspheres was 0.25 mg\/mL. Using a fluorescence reader, the minimum detection thresholds, expressed as the ratios of the fluorescence intensity of the test line to that of the control line (\n                      <jats:italic>T<\/jats:italic>\n                      \/\n                      <jats:italic>C<\/jats:italic>\n                      ), were determined to be 0.04109 for\n                      <jats:italic>Brucella<\/jats:italic>\n                      and 0.0276 for\n                      <jats:italic>M. bovis<\/jats:italic>\n                      . The dual test strip showed high specificity without detectable cross-reactivity, higher sensitivity than commercial colloidal gold test strips, and excellent intra- and inter-batch reproducibility (coefficients of variation &amp;lt;10%). Preliminary testing of 67 clinical samples showed that the dual test strip achieved agreement rates of 92.54% and 95.52% with commercial ELISA antibody detection kits for bovine brucellosis and bovine tuberculosis, respectively. A fluorescent microsphere-based dual-antibody rapid test strip was successfully developed for the simultaneous detection of antibodies against\n                      <jats:italic>Brucella<\/jats:italic>\n                      spp. and\n                      <jats:italic>M. bovis<\/jats:italic>\n                      . Exhibiting high sensitivity and specificity, this test strip serves as a highly suitable tool for the rapid diagnosis of bovine brucellosis and bovine tuberculosis antibodies.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fvets.2026.1881898","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T04:21:03Z","timestamp":1786422063000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a dual fluorescent microsphere\u2014based lateral flow immunoassays for simultaneous detection of Brucella and Mycobacterium bovis antibodies"],"prefix":"10.3389","volume":"13","author":[{"given":"Dihui","family":"Meng","sequence":"first","affiliation":[{"name":"College of Food Science and Engineering, Jilin Agricultural University","place":["Changchun, China"]},{"name":"Key Laboratory of Animal Parasitology of Ministry of Agriculture, Laboratory of Quality 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Products on Biohazards (Shanghai) of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangan","family":"Han","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Parasitology of Ministry of Agriculture, Laboratory of Quality and Safety Risk Assessment for Animal Products on Biohazards (Shanghai) of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Quan","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Parasitology of Ministry of Agriculture, Laboratory of Quality and Safety Risk Assessment for Animal Products on Biohazards (Shanghai) of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences","place":["Shanghai, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Ni","sequence":"additional","affiliation":[{"name":"College of Food Science and Engineering, Jilin Agricultural University","place":["Changchun, China"]},{"name":"Key Laboratory of Animal Parasitology of Ministry of Agriculture, Laboratory of Quality and Safety Risk Assessment for Animal Products on Biohazards (Shanghai) of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences","place":["Shanghai, China"]},{"name":"College of Life Sciences, Changchun Sci-Tech University","place":["Changchun, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Wu","sequence":"additional","affiliation":[{"name":"College of Food Science and Engineering, Jilin Agricultural University","place":["Changchun, China"]},{"name":"Key Laboratory of Animal Parasitology of Ministry of Agriculture, Laboratory of Quality and Safety Risk Assessment for Animal Products on Biohazards (Shanghai) of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences","place":["Shanghai, China"]},{"name":"College of Life Sciences, Changchun Sci-Tech University","place":["Changchun, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baoping","family":"Guo","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Xinjiang Medical University, and WHO-Collaborating Centre for Prevention and Care Management of Echinococcosis, The First Affiliated Hospital of Xinjiang Medical University","place":["Urumqi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lixin","family":"You","sequence":"additional","affiliation":[{"name":"College of Food Science and Engineering, Jilin Agricultural University","place":["Changchun, 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