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This study proposes a novel approach using immunochromatographic (IC) strip tests for the simultaneous detection of MTBC and NTM. Traditional methods for identifying mycobacteria, such as culture techniques, are hindered by delays in distinguishing between MTBC and NTM, which can affect patient care and disease control. Molecular methods, while sensitive, are resource-intensive and unable to differentiate between live and dead bacteria. In this research, we developed unique monoclonal antibodies (mAbs) against Ag85B, a mycobacterial secretory protein, and successfully implemented IC strip tests named 8B and 9B. These strips demonstrated high concordance rates with conventional methods for detecting MTBC, with positivity rates of 93.9% and 85.9%, respectively. For NTM detection, the IC strip tests achieved a 63.2% detection rate compared to culture methods, considering variations in growth rates among different NTM species. Furthermore, this study highlights a significant finding regarding the potential of MPT64 and Ag85B proteins as markers for MTBC detection. In conclusion, our breakthrough method enables rapid and accurate detection of both MTBC and NTM bacteria within the BACTEC MGIT system. This approach represents a valuable tool in clinical settings for distinguishing between MTBC and NTM infections, thereby enhancing the management and control of mycobacterial diseases.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Key points<\/jats:title>\n            <jats:p>\n              <jats:italic>\u2022 Panel of mAbs for differentiating MTB versus NTM<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\n              <jats:italic>\u2022 IC strips for diagnosing MTBC and NTM after the BACTEC MGIT<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\n              <jats:italic>\u2022 Combined detection of MTP64 and Ag85B enhances diagnostic accuracy<\/jats:italic>\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1007\/s00253-024-13293-1","type":"journal-article","created":{"date-parts":[[2024,9,2]],"date-time":"2024-09-02T10:02:17Z","timestamp":1725271337000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Rapid lateral flow test for Mycobacterium tuberculosis complex and non-tuberculous mycobacteria differentiation"],"prefix":"10.1007","volume":"108","author":[{"given":"Ponrut","family":"Phunpae","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weeraya","family":"Thongkum","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wutthichai","family":"Panyasit","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Witida","family":"Laopajon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nuchjira","family":"Takheaw","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Supansa","family":"Pata","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Umpa","family":"Yasamut","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Watchara","family":"Kasinrerk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4714-7945","authenticated-orcid":false,"given":"Chatchai","family":"Tayapiwatana","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,9,2]]},"reference":[{"key":"13293_CR1","doi-asserted-by":"publisher","first-page":"103552","DOI":"10.1016\/j.ebiom.2021.103552","volume":"71","author":"AR Anand","year":"2021","unstructured":"Anand AR, Biswas J (2021) TB or NTM: Can a new multiplex PCR assay be the answer? 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