{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:58:25Z","timestamp":1782280705506,"version":"3.54.5"},"posted":{"date-parts":[[2024,11,18]]},"group-title":"Microbiology","reference-count":59,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,11,18]],"date-time":"2024-11-18T00:00:00Z","timestamp":1731888000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,8,23]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Antibiotic-resistant bacteria are spreading in clinical, industrial, and environmental ecosystems. The spreading dynamics to and from the environment are unknown, largely due to the lack of appropriate (robust, fast, low-cost) analytical assays. In this study, we developed C12a, a versatile molecular toolbox to detect genetic markers of antibiotic resistance using CRISPR\/Cas12a. Biochemical characterization show that the C12a toolbox can detect less than 100 attoMolar of pure DNA fragments from the\n                  <jats:italic>blaCTX-M15<\/jats:italic>\n                  and\n                  <jats:italic>floR<\/jats:italic>\n                  genes, conferring resistance to b-lactams and amphenicols, respectively important for human and veterinary uses. In microbiological assays, C12a detected less than 10\n                  <jats:sup>2<\/jats:sup>\n                  CFU\/mL and high concordance was observed if compared to antibiotic susceptibility tests, PCR, or to whole genome sequencing. Additionally, C12a confirmed a high prevalence of the integrase\/integron system in\n                  <jats:italic>E. coli<\/jats:italic>\n                  isolates containing multiple antibiotic resistance genes (ARGs). The C12a toolbox shows equivalent detection performance in diverse laboratory settings, results redout (Fluorescence\n                  <jats:italic>vs<\/jats:italic>\n                  FLA) or input sample. Altogether, this work presents a comprehensive proof-of-concept, development description, and biochemical characterization of a collection of molecular tools to detect antibiotic resistance markers in a one health setup.\n                <\/jats:p>","DOI":"10.1101\/2024.11.14.623642","type":"posted-content","created":{"date-parts":[[2024,11,18]],"date-time":"2024-11-18T11:05:22Z","timestamp":1731927922000},"source":"Crossref","is-referenced-by-count":2,"title":["Versatile and Portable Cas12a-mediated Detection of Antibiotic Resistance Markers"],"prefix":"10.64898","author":[{"given":"Maryhory","family":"Vargas-Reyes","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2028-631X","authenticated-orcid":false,"given":"Roberto","family":"Alc\u00e1ntara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soraya","family":"Alfonsi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katherin","family":"Pe\u00f1aranda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dezemona","family":"Petrelli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberto","family":"Spurio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica J.","family":"Pajuelo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6679-5473","authenticated-orcid":false,"given":"Pohl","family":"Milon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025082602000500000_2024.11.14.623642v2.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2019.10.008"},{"key":"2025082602000500000_2024.11.14.623642v2.2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0804.010312"},{"key":"2025082602000500000_2024.11.14.623642v2.3","doi-asserted-by":"crossref","first-page":"100224","DOI":"10.1016\/j.coesh.2020.10.012","article-title":"Aspects of excessive antibiotic consumption and environmental influences correlated with the occurrence of resistance to antimicrobial agents","volume":"19","year":"2021","journal-title":"Curr Opin Environ Sci Health"},{"key":"2025082602000500000_2024.11.14.623642v2.4","doi-asserted-by":"publisher","DOI":"10.1021\/acs.est.5b02566"},{"key":"2025082602000500000_2024.11.14.623642v2.5","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-29283-8"},{"key":"2025082602000500000_2024.11.14.623642v2.6","doi-asserted-by":"crossref","unstructured":"von Wintersdorff, C. 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