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Unlike conventional molecular diagnostic methods that target only a single gene, which may lead to misdiagnosis or missed diagnosis, this CE-based multiplex approach provides comprehensive detection to reduce diagnostic errors. Specificity testing with 76 microorganisms representing common respiratory pathogens confirmed 100% analytical specificity with no cross-reactivity, while sensitivity analysis demonstrated detection limits ranging from 10 to 20 copies\/mL for all three target genes. In a prospective clinical validation study of 1067 patients suspected of pulmonary tuberculosis, the multiplex assay showed 77.4% sensitivity (CI 74.9%\u201379.9%), 99.6% specificity (CI 99.2%\u2013100%), 96.0% positive predictive value (CI 94.8%\u201397.2%), and 97.1% negative predictive value (CI 96.1%\u201398.1%). Notably, the study identified 6 MTBC strains (4.8% of TB patients) with\n                      <jats:italic>IS6110<\/jats:italic>\n                      deletions through whole-genome sequencing, which would result in false-negative results for any commercial PCR kits targeting\n                      <jats:italic>IS6110<\/jats:italic>\n                      . This integrated multiplex approach enhances diagnostic accuracy by simultaneously targeting multiple genes; then it offers the potential to reduce misdiagnosis and missed diagnosis of tuberculosis. In summary, the multiplex assay provides a more comprehensive alternative to current single-target molecular methods for MTBC detection.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Key points<\/jats:title>\n                    <jats:p>\n                      \u2022\n                      <jats:italic>The multiplex assay provides one-run results for IS6110, rpoB, and HSP65.<\/jats:italic>\n                    <\/jats:p>\n                    <jats:p>\n                      \u2022\u00a0\n                      <jats:italic>The multiplex assay is a more comprehensive method to detect MTBC.<\/jats:italic>\n                    <\/jats:p>\n                    <jats:p>\n                      \u2022\u00a0\n                      <jats:italic>This approach can reduce misdiagnosis and missed diagnosis of TB.<\/jats:italic>\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s00253-025-13701-0","type":"journal-article","created":{"date-parts":[[2026,2,2]],"date-time":"2026-02-02T19:52:32Z","timestamp":1770061952000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A multiplex assay based on capillary electrophoresis to detect Mycobacterium tuberculosis complex: development and clinical validation"],"prefix":"10.1007","volume":"110","author":[{"given":"Yaocheng","family":"Wang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhen","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li","family":"Lai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yiping","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yi","family":"Huang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,2,2]]},"reference":[{"issue":"6","key":"13701_CR1","doi-asserted-by":"publisher","DOI":"10.1002\/bio.4795","volume":"39","author":"M Batuer","year":"2024","unstructured":"Batuer M, Yuan Y, Yu M, Meng C (2024) Establishment and evaluation of a new fluorescent probe method based on loop-mediated isothermal amplification for the detection of Mycobacterium tuberculosis complex. 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