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Here, we propose a label-free method for Listeria monocytogenes subtype detection based on a hybrid split G-quadruplex (G4) and fusion recombinase-aided amplification (RAA)-cascaded CRISPR\/Cas13a (HGFRC) platform. Fusion RAA co-amplified species-specific and clonal complex (CC)-specific targets into 535 bp fusion products, which activated T7 transcription and Cas13a trans-cleavage activity. Activated Cas13a cleaved the RNA linker, disrupting G4 assembly and reducing fluorescence to generate a signal-off response. This design eliminated labeled probes, enabling single-signal detection. The assay achieved a sensitivity of 1.0 \u00d7 101 CFU\/mL within 45 min, with good specificity and sample suitability. Overall, this study provides a label-free, sensitive, and specific method for detecting L. monocytogenes CC5, offering new insights for bacterial subtype biosensing.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c09842","type":"journal-article","created":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T15:02:23Z","timestamp":1789657343000},"page":"30407-30417","source":"Crossref","is-referenced-by-count":0,"title":["DNA Fragment Fusion-Triggered\nLabel-Free Detection\nof\n                    <i>Listeria monocytogenes<\/i>\n                    Subtypes\nvia Fusion RAA and Hybrid Split G-Quadruplex"],"prefix":"10.1021","volume":"74","author":[{"given":"Fan","family":"Li","sequence":"first","affiliation":[{"name":"Shaanxi University of Science and Technology , , ,","place":["Xi\u2019an, China, 710021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui","family":"Han","sequence":"additional","affiliation":[{"name":"Shaanxi University of Science and Technology , , ,","place":["Xi\u2019an, China, 710021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruiting","family":"Yao","sequence":"additional","affiliation":[{"name":"Shaanxi University of Science and Technology , , ,","place":["Xi\u2019an, China, 710021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baoqing","family":"Zhou","sequence":"additional","affiliation":[{"name":"Jiangxi General Institute of Testing and Certification , ,","place":["Nanchang, China, 330052"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinxin","family":"Jiang","sequence":"additional","affiliation":[{"name":"Shaanxi University of Science and Technology , , ,","place":["Xi\u2019an, China, 710021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haorui","family":"Zhu","sequence":"additional","affiliation":[{"name":"Shaanxi University of Science and Technology , , ,","place":["Xi\u2019an, China, 710021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-0232-9251","authenticated-orcid":true,"given":"Yiheng","family":"Shi","sequence":"additional","affiliation":[{"name":"Shaanxi University of Science and Technology , , ,","place":["Xi\u2019an, China, 710021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8534-0914","authenticated-orcid":true,"given":"Lihua","family":"Fan","sequence":"additional","affiliation":[{"name":"Shaanxi University of Science and Technology , , ,","place":["Xi\u2019an, China, 710021"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5895-2237","authenticated-orcid":true,"given":"Di","family":"Wu","sequence":"additional","affiliation":[{"name":"Queen\u2019s University Belfast , , 19 Chlorine Gardens , ,","place":["Belfast, U.K., BT9 5DL"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7951-9750","authenticated-orcid":true,"given":"Guoliang","family":"Li","sequence":"additional","affiliation":[{"name":"Shaanxi University of Science and Technology , , ,","place":["Xi\u2019an, China, 710021"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,17]]},"reference":[{"key":"2026093009073735000_cit1","doi-asserted-by":"publisher","first-page":"112032","DOI":"10.1016\/j.foodcont.2026.112032","article-title":"Modelling Listeria monocytogenes contamination and application of\nthe resilience concept in the soft cheese supply chain","volume":"184","author":"Himani","year":"2026","journal-title":"Food Control"},{"issue":"7","key":"2026093009073735000_cit2","doi-asserted-by":"publisher","first-page":"1266","DOI":"10.3390\/foods14071266","article-title":"Listeria monocytogenes and Listeriosis: The global\nenigma","volume":"14","author":"Manyi-Loh","year":"2025","journal-title":"Foods"},{"key":"2026093009073735000_cit3","doi-asserted-by":"publisher","DOI":"10.1016\/j.jfutfo.2026.01.009","article-title":"Diversity shifts in human gut microbiota induced by Listeria monocytogenes: An in vitro co-culture study","author":"Lu","year":"2026","journal-title":"J. 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