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The suboptimal\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    \u2013expressed recombinant proteins limit the diagnostic performance of current bTB\u00a0serological tests. To overcome this limitation, we evaluated\n                    <jats:italic>Mycolicibacterium smegmatis<\/jats:italic>\n                    as an expression host capable of producing recombinant antigens with post-translational modifications comparable to those of\n                    <jats:italic>M. bovis<\/jats:italic>\n                    . Ten antigen candidates were individually expressed in\n                    <jats:italic>E. coli<\/jats:italic>\n                    using the pET-26b(\u2009+) vector and in\n                    <jats:italic>M. smegmatis<\/jats:italic>\n                    using the pSO\u0394Bam vector. Their diagnostic performance was evaluated using an\u00a0enzyme-linked immunosorbent assay\u00a0(ELISA) with plasma samples from interferon-gamma release assay (IGRA)-negative (\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u200930) and -positive (\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u200946) cattle in South Korea, followed by receiver operating characteristic (ROC) curve analysis. Among the single antigens, LprA expressed in\n                    <jats:italic>M. smegmatis<\/jats:italic>\n                    demonstrated diagnostic performance comparable to that of\u00a0the well-established antigen MPB70 (sensitivity: 50.0%, specificity: 96.6%, AUC: 0.791). In addition, several\n                    <jats:italic>M. smegmatis<\/jats:italic>\n                    \u2013derived antigens showed higher concordance with the IGRA results, as assessed by Cohen\u2019s kappa and Fisher\u2019s exact tests, and a stronger association between age and antigen-specific antibody responses was observed among IGRA-positive cattle. Moreover, a multiple logistic regression model incorporating eight antigens, including those derived from both hosts, achieved high predictive accuracy for IGRA results (sensitivity: 87.0%, specificity: 100%, AUC: 0.991). These findings suggest that\n                    <jats:italic>M. smegmatis<\/jats:italic>\n                    is a promising host for identifying novel antigens and that a multi-host strategy may improve bTB serodiagnosis.\n                  <\/jats:p>","DOI":"10.1186\/s13567-026-01743-9","type":"journal-article","created":{"date-parts":[[2026,5,11]],"date-time":"2026-05-11T09:03:46Z","timestamp":1778490226000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Recombinant proteins expressed in Mycolicibacterium smegmatis enhance antibody-based detection of bovine tuberculosis"],"prefix":"10.1186","volume":"57","author":[{"given":"Eomseob","family":"Jang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kilhan","family":"Kwak","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Norikazu","family":"Isoda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kazuhiro","family":"Matsuo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeewan","family":"Thapa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chie","family":"Nakajima","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yasuhiko","family":"Suzuki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,5,8]]},"reference":[{"key":"1743_CR1","unstructured":"Global tuberculosis report (2024) https:\/\/www.who.int\/publications\/i\/item\/9789240101531. 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