{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T14:32:19Z","timestamp":1787841139484,"version":"build-2784847793"},"reference-count":21,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"1","license":[{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:sec>\n                    <jats:title>Background:<\/jats:title>\n                    <jats:p>\n                      Tuberculosis (TB) is regarded as one of the most challenging infectious diseases globally, posing significant obstacles in terms of diagnosis and treatment. The limited effectiveness of the Bacillus Calmette\u2013Gu\u00e9rin vaccine and the low sensitivity of conventional diagnostic methods underscore the need for improved antigen-based detection strategies. This study aimed to clone, express, and characterize culture filtrate protein 10 (CFP-10) and resuscitation-promoting factor D (rpfD) and to predict potential T-cell and B-cell epitopes using\n                      <jats:italic toggle=\"yes\">in silico<\/jats:italic>\n                      methods.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods:<\/jats:title>\n                    <jats:p>\n                      Genes encoding Rv3874 (CFP-10) and Rv2389c (rpfD) were amplified from\n                      <jats:italic toggle=\"yes\">Mycobacterium tuberculosis<\/jats:italic>\n                      H37Rv, subsequently cloned into the pCold II vector and then expressed in\n                      <jats:italic toggle=\"yes\">Escherichia coli<\/jats:italic>\n                      BL21 (DE3). Induction with isopropylthio-\u03b2-galactoside resulted in recombinant protein expression and confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).\n                      <jats:italic toggle=\"yes\">In silico<\/jats:italic>\n                      analysis included physicochemical characterization, secretion and localization prediction, and identification of T-cell and B-cell epitopes, followed by screening for antigenicity, allergenicity, and toxicity.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results:<\/jats:title>\n                    <jats:p>Polymerase chain reaction successfully produced fragments of 303 bp (Rv3874) and 465 bp (Rv2389c), and sequencing confirmed the correct construct. SDS-PAGE revealed recombinant proteins of ~11 kDa (CFP-10) and ~15\u201316 kDa (rpfD), consistent with theoretical sizes. Both proteins were predicted to be antigenic and extracellular. Multiple strong binding epitopes were identified with high antigenicity, which were nontoxic and nonallergenic which signal a large immunogenic capacity.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions:<\/jats:title>\n                    <jats:p>\n                      CFP-10 and rpfD were successfully cloned and expressed and demonstrated promising immunogenic profiles\n                      <jats:italic toggle=\"yes\">in silico<\/jats:italic>\n                      . These findings support the potential as complementary antigens in future vaccine platforms and immunodiagnostic tools for TB.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.4103\/ijmy.ijmy_11_26","type":"journal-article","created":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T20:00:17Z","timestamp":1774641617000},"page":"31-37","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":1,"title":["Molecular Cloning and In silico Epitope Prediction of Culture Filtrate Protein 10 and Resuscitation-promoting Factor D from Mycobacterium tuberculosis with Potential Immunodiagnostic Applications"],"prefix":"10.4103","volume":"15","author":[{"given":"Fadhilah Ananda","family":"Putri","sequence":"first","affiliation":[{"name":"Department of Biomedical Sciences, Graduate School, Hasanuddin University, Makassar, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3347-6529","authenticated-orcid":false,"given":"Muhammad Nasrum","family":"Massi","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rizalinda","family":"Sjahril","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahyar","family":"Ahmad","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Mathematics and Nature Science, Hasanuddin University, Makassar, Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mochammad","family":"Hatta","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of 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