{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T19:53:18Z","timestamp":1789501998322,"version":"build-2803163510"},"reference-count":66,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,1,31]],"date-time":"2024-01-31T00:00:00Z","timestamp":1706659200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100006190","name":"Research and Development","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100006190","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>\n                    The principal pathogen responsible for chronic urinary tract infections, immunocompromised hosts, and cystic fibrosis patients is\n                    <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                    , which is difficult to eradicate. Due to the extensive use of antibiotics, multidrug-resistant\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    has evolved, complicating clinical therapy. Therefore, a rapid and efficient approach for detecting\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    strains and their resistance genes is necessary for early clinical diagnosis and appropriate treatment. This study combines recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats-association protein 13a (CRISPR-Cas13a) to establish a one-tube and two-step reaction systems for detecting the\n                    <jats:italic>mexX<\/jats:italic>\n                    gene in\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    . The test times for one-tube and two-step RPA-Cas13a methods were 5 and 40\u2009min (including a 30\u2009min RPA amplification reaction), respectively. Both methods outperform Quantitative Real-time Polymerase Chain Reactions (qRT-PCR) and traditional PCR. The limit of detection (LoD) of\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    genome in one-tube and two-step RPA-Cas13a is 10 aM and 1 aM, respectively. Meanwhile, the designed primers have a high specificity for\n                    <jats:italic>P. aeruginosa mexX<\/jats:italic>\n                    gene. These two methods were also verified with actual samples isolated from industrial settings and demonstrated great accuracy. Furthermore, the results of the two-step RPA-Cas13a assay could also be visualized using a commercial lateral flow dipstick with a LoD of 10 fM, which is a useful adjunt to the gold-standard qRT-PCR assay in field detection. Taken together, the procedure developed in this study using RPA and CRISPR-Cas13a provides a simple and fast way for detecting resistance genes.\n                  <\/jats:p>","DOI":"10.3389\/fmicb.2024.1341179","type":"journal-article","created":{"date-parts":[[2024,1,30]],"date-time":"2024-01-30T23:43:18Z","timestamp":1706658198000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":13,"title":["Rapid detection of mexX in Pseudomonas aeruginosa based on CRISPR-Cas13a coupled with recombinase polymerase amplification"],"prefix":"10.3389","volume":"15","author":[{"given":"Xiao-Xuan","family":"Zhu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying-Si","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Su-Juan","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ru-Qun","family":"Peng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xia","family":"Wen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Peng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qing-Shan","family":"Shi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gang","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao-Bao","family":"Xie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,1,31]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"108084","DOI":"10.1016\/j.envint.2023.108084","article-title":"A global baseline for qPCR-determined antimicrobial resistance gene prevalence across environments","volume":"178","author":"Abramova","year":"2023","journal-title":"Environ. 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