{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T22:50:33Z","timestamp":1791931833089,"version":"4.3.4"},"reference-count":37,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T00:00:00Z","timestamp":1713225600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000704","name":"Royal Society of Chemistry","doi-asserted-by":"publisher","award":["R20-7704"],"award-info":[{"award-number":["R20-7704"]}],"id":[{"id":"10.13039\/501100000704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000704","name":"Royal Society of Chemistry","doi-asserted-by":"publisher","award":["ILLU-698-353"],"award-info":[{"award-number":["ILLU-698-353"]}],"id":[{"id":"10.13039\/501100000704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000704","name":"Royal Society of Chemistry","doi-asserted-by":"publisher","award":["ILLU-698-930"],"award-info":[{"award-number":["ILLU-698-930"]}],"id":[{"id":"10.13039\/501100000704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005825","name":"U.S. Department of Agriculture NIFA","doi-asserted-by":"publisher","award":["R20-7704"],"award-info":[{"award-number":["R20-7704"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005825","name":"U.S. Department of Agriculture NIFA","doi-asserted-by":"publisher","award":["ILLU-698-353"],"award-info":[{"award-number":["ILLU-698-353"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005825","name":"U.S. Department of Agriculture NIFA","doi-asserted-by":"publisher","award":["ILLU-698-930"],"award-info":[{"award-number":["ILLU-698-930"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Antimicrobial-resistant (AMR) bacteria pose a significant global health threat, and bacteria that produce New Delhi metallo-\u03b2-lactamase (NDM) are particularly concerning due to their resistance to most \u03b2-lactam antibiotics, including carbapenems. The emergence and spread of NDM-producing genes in food-producing animals highlight the need for a fast and accurate method for detecting AMR bacteria. We therefore propose a PCR-coupled CRISPR\/Cas12a-based fluorescence assay that can detect NDM-producing genes (blaNDM) in bacteria. Thanks to its designed gRNA, this CRISPR\/Cas12a system was able to simultaneously cleave PCR amplicons and ssDNA-FQ reporters, generating fluorescence signals. Our method was found to be highly specific when tested against other foodborne pathogens that do not carry blaNDM and also demonstrated an excellent capability to distinguish single-nucleotide polymorphism. In the case of blaNDM-1 carrying E. coli, the assay performed exceptionally well, with a detection limit of 2.7 \u00d7 100 CFU\/mL: 100 times better than conventional PCR with gel electrophoresis. Moreover, the developed assay detected AMR bacteria in food samples and exhibited enhanced performance compared to previously published real-time PCR assays. Thus, this novel PCR-coupled CRISPR\/Cas12a-based fluorescence assay has considerable potential to improve current approaches to AMR gene detection and thereby contribute to mitigating the global threat of AMR.<\/jats:p>","DOI":"10.3390\/bios14040194","type":"journal-article","created":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T11:13:02Z","timestamp":1713265982000},"page":"194","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":35,"title":["A CRISPR\/Cas12a-Based System for Sensitive Detection of Antimicrobial-Resistant Genes in Carbapenem-Resistant Enterobacterales"],"prefix":"10.3390","volume":"14","author":[{"given":"Jiyong","family":"Shin","sequence":"first","affiliation":[{"name":"Department of Food Science and Human Nutrition, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sei Rim","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Food Science and Human Nutrition, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-8904-2552","authenticated-orcid":false,"given":"Zifan","family":"Xie","sequence":"additional","affiliation":[{"name":"Department of Food Science and Human Nutrition, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4464-9536","authenticated-orcid":false,"given":"Yong-Su","family":"Jin","sequence":"additional","affiliation":[{"name":"Department of Food Science and Human Nutrition, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi-Cheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Food Science and Human Nutrition, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA"},{"name":"Center for Digital Agriculture, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,4,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3903","DOI":"10.2147\/IDR.S234610","article-title":"Antimicrobial resistance: Implications and costs","volume":"12","author":"Dadgostar","year":"2019","journal-title":"Infect. 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