{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T17:42:16Z","timestamp":1790703736397,"version":"4.1.0"},"reference-count":38,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,8,21]],"date-time":"2024-08-21T00:00:00Z","timestamp":1724198400000},"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. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p><jats:italic>Mycobacterium tuberculosis<\/jats:italic>, the causative agent of human tuberculosis, poses a significant threat to global public health and imposes a considerable burden on the economy. However, existing laboratory diagnostic methods for <jats:italic>M. tuberculosis<\/jats:italic> are time-consuming and have limited sensitivity levels.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The CRISPR\/Cas system, commonly known as the \u201cgene scissors\u201d, demonstrates remarkable specificity and efficient signal amplification capabilities. Enzymatic recombinase amplification (ERA) was utilized to rapidly amplify trace DNA fragments at a consistent temperature without relying on thermal cyclers. By integrating of CRISPR\/Cas12a with ERA, we successfully developed an ERA-CRISPR\/Cas12a detection system that enables rapid identification of <jats:italic>M. tuberculosis<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The sensitivity of the ERA-CRISPR\/Cas12a fluorescence and lateral flow systems was 9 copies\/\u03bcL and 90 copies\/\u03bcL, respectively. Simultaneously, the detection system exhibited no cross-reactivity with various of respiratory pathogens and non-tuberculosis mycobacteria, demonstrating a specificity of 100%. The positive concordance rate between the ERA-CRISPR\/Cas12a fluorescence system and commercial qPCR was 100% in 60 clinical samples. Meanwhile, the lateral flow system showed a positive concordance rate of 93.8% when compared to commercial qPCR. Both methods demonstrated a negative concordance rate of 100%, and the test results can be obtained in 50 min at the earliest.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>The ERA-CRISPR\/Cas12a system offers a rapid, sensitive, and specific method that presents a novel approach to laboratory diagnosis of <jats:italic>M. tuberculosis<\/jats:italic>.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2024.1454076","type":"journal-article","created":{"date-parts":[[2024,8,23]],"date-time":"2024-08-23T10:01:13Z","timestamp":1724407273000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":21,"title":["ERA-CRISPR\/Cas12a system: a rapid, highly sensitive and specific assay for Mycobacterium tuberculosis"],"prefix":"10.3389","volume":"14","author":[{"given":"Tian","family":"Gan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianwei","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongliang","family":"Deng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"He","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,8,21]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1361","DOI":"10.1080\/22221751.2019.1664939","article-title":"CRISPR-based rapid and ultra-sensitive diagnostic test for Mycobacterium tuberculosis","volume":"8","author":"Ai","year":"2019","journal-title":"Emerg. 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