{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T15:16:52Z","timestamp":1776784612726,"version":"3.51.2"},"reference-count":27,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,3,7]],"date-time":"2025-03-07T00:00:00Z","timestamp":1741305600000},"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. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>A new class of <jats:italic>Mycobacterium tuberculosis<\/jats:italic> (MTB) antigen-based skin tests was recommended by WHO for the diagnosis of TB infection. However, their performance in some settings remains suboptimal. Our study focused on screening novel MTB recombinant antigens for skin tests and evaluating their preclinical efficacy for TB infection detection.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We constructed <jats:italic>Escherichia coli<\/jats:italic> BL21 expression vectors to produce a series of recombinant MTB antigens. We assessed their ability to detect tuberculosis infection through skin tests. Model animals sensitized with MTB and BCG were treated with a total of 24 MTB recombinant antigens.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Out of 24 tested recombinant MTB antigens, only three (E-M, E-C-M, and E-7.7-C) met the criteria for skin test reaction interpretation for preclinical trials. Among these, the mean values of skin reaction from E-M were found to be comparable to EC (P&amp;gt;0.05), with no cross-reaction with BCG. Additionally, E-M exhibited a strong safety preclinical profile with no significant abnormalities in physiological, biochemical, or histopathological assessments, supporting its suitability for clinical evaluation. It also displayed high specificity by differentiating MTB infection from BCG vaccination and NTM infection, with no cross-reactivity observed in sensitized guinea pigs.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>Our results indicate that the E-M recombinant antigen possesses promising characteristics for the detection of tuberculosis infection, demonstrating good safety and efficacy at the preclinical level. Further clinical trials are required to assess its clinical safety, efficacy, and feasibility.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1498448","type":"journal-article","created":{"date-parts":[[2025,3,7]],"date-time":"2025-03-07T05:23:09Z","timestamp":1741324989000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Screening and preclinical assessment of novel Mycobacterium tuberculosis recombinant antigens based tuberculin skin testing"],"prefix":"10.3389","volume":"16","author":[{"given":"Xichao","family":"Ou","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eryong","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Faiqa","family":"Rashid","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shaojun","family":"Pei","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guoqin","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard","family":"Anthony","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruida","family":"Xing","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fei","family":"Huang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hui","family":"Xia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yang","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuanyuan","family":"Song","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yang","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bing","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shengfen","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Minggui","family":"Lin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lin","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanlin","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,3,7]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1101\/2024.01.02.23300347","article-title":"A novel comprehensive Tuberculosis (TB) control programme methodology based on the nexus of participatory action research inspired public health and precision treatment approach","volume":"4","author":"Mitra","year":"2024","journal-title":"medRxiv"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijid.2020.02.042","article-title":"Tools to implement the World Health Organization End TB Strategy: Addressing common challenges in high and low endemic countries","volume":"92","author":"Al Abri","year":"2020","journal-title":"Int J Infect Dis"},{"key":"B3","volume-title":"Gear up to end TB: introducing the end TB strategy","year":"2015"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(15)60570-0","article-title":"WHO\u2019s new end TB strategy","volume":"385","author":"Uplekar","year":"2015","journal-title":"Lancet"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1093\/ofid\/ofad228","article-title":"A systematic review on the safety of mycobacterium tuberculosis\u2013specific antigen\u2013based skin tests for tuberculosis infection compared with tuberculin skin tests","volume":"10","author":"Hamada","year":"2023","journal-title":"Open Forum Infect Dis"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1162177","article-title":"Utility of recombinant fusion protein ESAT6-CFP10 skin test for differential diagnosis of active tuberculosis: A prospective study","volume":"14","author":"Yuan","year":"2023","journal-title":"Front Immunol"},{"key":"B7","volume-title":"WHO consolidated guidelines on tuberculosis Module 2: Screening\u2013Systematic screening for tuberculosis disease","author":"MacPherson","year":"2021"},{"key":"B8","volume-title":"WHO consolidated guidelines on tuberculosis. Module 3: diagnosis-rapid diagnostics for tuberculosis detection","year":"2020"},{"key":"B9","volume-title":"WHO consolidated guidelines on tuberculosis. Module 3: diagnosis. Tests for TB infection","year":"2022"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(21)00261-9","article-title":"The diagnostic performance of novel skin-based in-vivo tests for tuberculosis infection compared with purified protein derivative tuberculin skin tests and blood-based in vitro interferon-\u03b3 release assays: a systematic review and meta-analysis","volume":"22","author":"Krutikov","year":"2022","journal-title":"Lancet Infect Dis"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1016\/bs.acc.2023.03.003","article-title":"Advances in TB testing","volume":"115","author":"Pagaduan","year":"2023","journal-title":"Adv Clin Chem"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.72.5.2574-2581.2004","article-title":"Evaluation of T-cell responses to novel RD1-and RD2-encoded Mycobacterium tuberculosis gene products for specific detection of human tuberculosis infection","volume":"72","author":"Liu","year":"2004","journal-title":"Infection Immun"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1007\/s40291-015-0157-0","article-title":"Novel T-cell assays for the discrimination of active and latent tuberculosis infection: the diagnostic value of PPE family","volume":"19","author":"Pourakbari","year":"2015","journal-title":"Mol diagnosis Ther"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1128\/iai.63.12.4613-4618.1995","article-title":"Mapping of the delayed-type hypersensitivity-inducing epitope of secreted protein MPT64 from Mycobacterium tuberculosis","volume":"63","author":"Oettinger","year":"1995","journal-title":"Infection Immun"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1016\/s0962-8479(96)90009-x","article-title":"Clinical evaluation of MPT-64 and MPT-59, two proteins secreted from Mycobacterium tuberculosis, for skin test reagents","volume":"77","author":"Wilcke","year":"1996","journal-title":"Tuber Lung Dis"},{"key":"B16","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1128\/CVI.00481-12","article-title":"Mycobacterium tuberculosis region of difference (RD) 2 antigen Rv1985c and RD11 antigen Rv3425 have the promising potential to distinguish patients with active tuberculosis from M. bovis BCG-vaccinated individuals","volume":"20","author":"Wang","year":"2013","journal-title":"Clin Vaccine Immunol"},{"key":"B17","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.clim.2018.05.002","article-title":"23-valent pneumococcal polysaccharide vaccine elicits hierarchical antibody and cellular responses in healthy and tuberculosis-cured elderly, and HIV-1-infected subjects","volume":"193","author":"Huang","year":"2018","journal-title":"Clin Immunol"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1080\/23744235.2017.1410282","article-title":"Diagnostic usefulness of the QuantiFERON-TB gold in-tube test (QFT-GIT) for tuberculous vertebral osteomyelitis","volume":"50","author":"Choi","year":"2018","journal-title":"Infect Dis"},{"key":"B19","doi-asserted-by":"publisher","first-page":"105002","DOI":"10.1016\/j.micinf.2022.105002","article-title":"Novel serological biomarker panel using protein microarray can distinguish active TB from latent TB infection","volume":"24","author":"Li","year":"2022","journal-title":"Microbes Infection"},{"key":"B20","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1186\/s13568-020-01068-6","article-title":"The single recombinant M. tuberculosis protein DPPD provides enhanced performance of skin testing among HIV-infected tuberculosis patients","volume":"10","author":"Badaro","year":"2020","journal-title":"Amb Express"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.01464-2015","article-title":"Sensitivity of C-Tb: a novel RD-1-specific skin test for the diagnosis of tuberculosis infection","volume":"47","author":"Hoff","year":"2016","journal-title":"Eur Respir J"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0208705","article-title":"Comparative sensitivity of the test with tuberculosis recombinant allergen, containing ESAT6-CFP10 protein, and Mantoux test with 2 TU PPD-L in newly diagnosed tuberculosis children and adolescents in Moscow","volume":"13","author":"Slogotskaya","year":"2018","journal-title":"PloS One"},{"key":"B23","article-title":"Qualitative evidence for the use of Mycobacterium tuberculosis antigen-based skin tests","author":"Chan","year":"2022"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1086\/jid.2000.182.issue-1","article-title":"Cloning of a Mycobacterium tuberculosis gene encoding a purifed protein derivative protein that elicits strong tuberculosis-specific delayed-type hypersensitivity","volume":"182","author":"Coler","year":"2000","journal-title":"J Infect Dis"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.5.1274-1282.1996","article-title":"Molecular analysis of genetic differences between Mycobacterium bovis BCG and virulent M. bovis","volume":"178","author":"Mahairas","year":"1996","journal-title":"J bacteriology"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.05995-11","article-title":"Performance of the MGIT TBc identification test and meta-analysis of MPT64 assays for identification of the Mycobacterium tuberculosis complex in liquid culture","volume":"49","author":"Brent","year":"2011","journal-title":"J Clin Microbiol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.7.3454-3456.1998","article-title":"Delayed-type hypersensitivity responses to ESAT-6 and MPT64 from Mycobacterium tuberculosis in the Guinea pig","volume":"66","author":"Elhay","year":"1998","journal-title":"Infection Immun"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1498448\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,7]],"date-time":"2025-03-07T05:23:17Z","timestamp":1741324997000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1498448\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,7]]},"references-count":27,"alternative-id":["10.3389\/fimmu.2025.1498448"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1498448","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,7]]},"article-number":"1498448"}}