{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T14:50:46Z","timestamp":1778856646954,"version":"3.51.4"},"reference-count":21,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,1,3]],"date-time":"2025-01-03T00:00:00Z","timestamp":1735862400000},"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. Microbiol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Timely and accurate diagnosis is crucial for the effective treatment and prevention of brucellosis. Current serological diagnostics, primarily based on lipopolysaccharide (LPS), suffer from cross-reactivity with other Gram-negative bacteria, which limits their specificity. Periplasmic protein 26 (BP26), a highly immunogenic antigen found in <jats:italic>Brucella<\/jats:italic>, has emerged as a promising alternative for enhancing diagnostic specificity. This study aimed to develop and evaluate a competitive enzyme-linked immunosorbent assay (C-ELISA) utilizing monoclonal antibodies against BP26 for the diagnosis of human brucellosis, thereby providing a more accurate and specific diagnostic approach.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The study produced monoclonal antibody (mAb) against the BP26 protein through traditional mouse hybridoma technology and developed the C-ELISA method, and compared with a C-ELISA method based on LPS mAb. The detection performance was validated through the analysis of 190 human serum samples, which included 95 brucellosis serum samples and 95 negative serum samples collected by the Xuzhou Center for Disease Control and Prevention, and a comparative analysis was conducted on the diagnostic efficacy of indirect ELISA for brucellosis using both BP26 and LPS-based methods.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The BP26 mAb based C-ELISA achieved 100% sensitivity and specificity in detecting human brucellosis, significantly outperforming the C-ELISA based LPS mAb. Furthermore, the accuracy of the indirect enzyme-linked immunosorbent assay (I-ELISA) using BP26 protein was 98.95%, compared to an accuracy of LPS diagnosis was 99.47%. These results indicated that the BP26 mAb can effectively and accurately detected human brucellosis infections.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This study successfully developed and evaluated a BP26 protein-based C-ELISA method for diagnosing human brucellosis, establishing a foundation for identifying alternative diagnostic antigens for brucellosis.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmicb.2024.1516915","type":"journal-article","created":{"date-parts":[[2025,1,3]],"date-time":"2025-01-03T06:47:35Z","timestamp":1735886855000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":7,"title":["Development and evaluation of the recombinant BP26 protein-based C-ELISA for human brucellosis diagnosis"],"prefix":"10.3389","volume":"15","author":[{"given":"Yujia","family":"Xie","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shaoqing","family":"Lin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Liping","family":"Guo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xinru","family":"Qi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shiqi","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qichuan","family":"Pei","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yixiao","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qi","family":"Wu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yun","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meixue","family":"Yao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dehui","family":"Yin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,1,3]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1080\/23144599.2024.2384015","article-title":"Seroprevalence and molecular detection of Brucella infection in livestock in the United Arab Emirates.","volume":"12","author":"Ameni","year":"2024","journal-title":"Int. J. Vet. Sci. Med."},{"key":"B2","first-page":"264","article-title":"Cloning and expression of B. mellitensis bp26 gene in Lactococcus lactis as a food grade vaccine.","volume":"11","author":"Azizpour Maghvan","year":"2019","journal-title":"Avicenna J. Med. Biotechnol."},{"key":"B3","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1016\/j.bbrc.2021.04.127","article-title":"Comparative analysis of the main outer membrane proteins of Brucella in the diagnosis of brucellosis.","volume":"560","author":"Bai","year":"2021","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"B4","doi-asserted-by":"publisher","first-page":"772","DOI":"10.1128\/cdli.8.4.772-775.2001","article-title":"Use of recombinant BP26 protein in serological diagnosis of Brucella melitensis infection in sheep.","volume":"8","author":"Cloeckaert","year":"2001","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"B5","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1016\/0378-1097(96)00169-3","article-title":"Cloning, nucleotide sequence, and expression of the Brucella melitensis bp26 gene coding for a protein immunogenic in infected sheep.","volume":"140","author":"Cloeckaert","year":"1996","journal-title":"FEMS Microbiol. Lett."},{"key":"B6","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1016\/j.medmal.2018.12.004","article-title":"Limitations of diagnostic tests for bacterial infections.","volume":"49","author":"Eldin","year":"2019","journal-title":"Med. Mal. Infect."},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2024.1389728","article-title":"Production and evaluation of anti-BP26 monoclonal antibodies for the serological detection of animal brucellosis.","volume":"11","author":"Guo","year":"2024","journal-title":"Front. Vet. Sci."},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1093\/femspd\/ftaa002","article-title":"A combined subunit vaccine comprising BP26, Omp25 and L7\/L12 against brucellosis.","volume":"77","author":"Gupta","year":"2019","journal-title":"Pathog. Dis."},{"key":"B9","doi-asserted-by":"publisher","first-page":"1789","DOI":"10.3201\/eid2909.230052","article-title":"Global estimate of human brucellosis incidence.","volume":"29","author":"Laine","year":"2023","journal-title":"Emerg. Infect. Dis."},{"key":"B10","doi-asserted-by":"publisher","first-page":"549","DOI":"10.1093\/jac\/dkl257","article-title":"Intracellular killing of Brucella melitensis in human macrophages with microsphere-encapsulated gentamicin.","volume":"58","author":"Lec\u00e1roz","year":"2006","journal-title":"J. Antimicrob. Chemother."},{"key":"B11","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1099\/0022-1317-51-1-20","article-title":"Induction of a th1-type of immune response but not protective immunity by intramuscular DNA immunisation with Brucella abortus GroEL heat-shock gene.","volume":"51","author":"Leclerq","year":"2002","journal-title":"J. Med. Microbiol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"719","DOI":"10.1139\/cjm-2017-0179","article-title":"Brucella melitensis M5-90\u0394bp26 as a potential live vaccine that allows for the distinction between natural infection and immunization.","volume":"63","author":"Li","year":"2017","journal-title":"Can. J. Microbiol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"1141","DOI":"10.1517\/13543784.15.10.1141","article-title":"Future trends in human brucellosis treatment.","volume":"15","author":"Pappas","year":"2006","journal-title":"Expert Opin. Investig. Drugs"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0034246","article-title":"Characterization of periplasmic protein BP26 epitopes of Brucella melitensis reacting with murine monoclonal and sheep antibodies.","volume":"7","author":"Qiu","year":"2012","journal-title":"PLoS One"},{"key":"B15","first-page":"776","article-title":"Rapid detection of Brucella by an automated blood culture system at a tertiary care hospital of north India.","volume":"139","author":"Raj","year":"2014","journal-title":"Indian J. Med. Res."},{"key":"B16","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1128\/jcm.34.1.165-169.1996","article-title":"Cloning of Brucella abortus gene and characterization of expressed 26-kilodalton periplasmic protein: potential use for diagnosis.","volume":"34","author":"Rossetti","year":"1996","journal-title":"J. Clin. Microbiol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"4326","DOI":"10.1128\/iai.71.8.4326-4332.2003","article-title":"Characterization of new members of the group 3 outer membrane protein family of Brucella spp.","volume":"71","author":"Salhi","year":"2003","journal-title":"Infect. Immun."},{"key":"B18","doi-asserted-by":"publisher","first-page":"305","DOI":"10.1128\/cdli.3.3.305-308.1996","article-title":"Enzyme-linked immunosorbent assay with partially purified cytosoluble 28-kilodalton protein for serological differentiation between Brucella melitensis-infected and B. melitensis Rev.1-vaccinated sheep.","volume":"3","author":"Salih-Alj Debbarh","year":"1996","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2022.106663","article-title":"Study on antigenic protein Omp2b in combination with Omp31 and BP26 for serological detection of human brucellosis.","volume":"205","author":"Wu","year":"2023","journal-title":"J. Microbiol. Methods"},{"key":"B20","doi-asserted-by":"publisher","first-page":"5301","DOI":"10.2147\/idr.S374432","article-title":"Comparison of BP26, Omp25 and Omp31 and a multiepitope-based fusion protein in the serological detection of canine brucellosis.","volume":"15","author":"Yao","year":"2022","journal-title":"Infect. Drug Resist."},{"key":"B21","doi-asserted-by":"publisher","first-page":"2001","DOI":"10.3201\/eid1612.091081","article-title":"Human brucellosis, Inner Mongolia, China.","volume":"16","author":"Zhang","year":"2010","journal-title":"Emerg. Infect. Dis."}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2024.1516915\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,1,3]],"date-time":"2025-01-03T06:47:38Z","timestamp":1735886858000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2024.1516915\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,3]]},"references-count":21,"alternative-id":["10.3389\/fmicb.2024.1516915"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2024.1516915","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,3]]},"article-number":"1516915"}}