{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T19:17:39Z","timestamp":1789154259491,"version":"build-2803163510"},"reference-count":38,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T00:00:00Z","timestamp":1779667200000},"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>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>The rise of Klebsiella pneumoniae (KP) and carbapenem-resistant KP (CRKP) poses grave threats to public health and food safety, creating an urgent demand for rapid point-of-care testing (POCT). Traditional detection methods are limited by laboratory barriers, making them unsuitable for POCT implementation.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Herein, a one-tube assay integrating recombinant polymerase amplification (RPA) with CRISPR\/Cas12a technology was developed for the rapid, sensitive and specific detection of KP and\n                      <jats:italic>blaOXA-48<\/jats:italic>\n                      -carrying CRKP. Specific primers targeting the KP-specific rpoB gene and carbapenem-resistance gene\n                      <jats:italic>blaOXA-48<\/jats:italic>\n                      were designed, and optimal primer pairs were screened via agarose gel electrophoresis. CrRNA sequences were designed according to RPA amplicons, and the components of the CRISPR\/Cas12a reaction were optimized. A two-step reaction system was initially evaluated, followed by the establishment of an integrated one-tube RPA-CRISPR\/Cas12a assay. A total of 66 clinical specimens and artificially contaminated food samples were used for method validation, with microbial culture and qPCR as reference methods.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The two-step assay was capable of detecting bacterial suspensions at a concentration of 100 CFU\/mL. The one-tube system could be completed within 1 hour at 37 \u00b0C. This assay avoided aerosol contamination and allowed visual result readout under blue light. In the validation test, the detection results of the one-tube assay were consistent with those obtained by microbial culture and qPCR.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      This study constructed a dual-target RPA-CRISPR\/Cas12a platform for the visual detection of KP and\n                      <jats:italic>blaOXA-48<\/jats:italic>\n                      -positive CRKP under blue light. This assay reduces reliance on sophisticated equipment and professional personnel. It can serve as a promising POCT tool for clinical diagnosis and food safety surveillance, and provides evidence for the timely formulation of rational antimicrobial treatment strategies.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2026.1817859","type":"journal-article","created":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T06:05:59Z","timestamp":1779689159000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Visual detection platform based on RPA-CRISPR\/Cas12a for Klebsiella pneumoniae and Carbapenem-resistant Klebsiella pneumoniae in clinical and food safety settings"],"prefix":"10.3389","volume":"16","author":[{"given":"Tuo","family":"Ji","sequence":"first","affiliation":[{"name":"Lianyungang Clinical College, Bengbu Medical University & The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]},{"name":"College of Life Sciences, Bengbu Medical University","place":["Bengbu, China"]},{"name":"Department of Central Laboratory, The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Fang","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Bengbu Medical University","place":["Bengbu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"YuZhi","family":"Gao","sequence":"additional","affiliation":[{"name":"Lianyungang Clinical College, Bengbu Medical University & The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]},{"name":"College of Life Sciences, Bengbu Medical University","place":["Bengbu, China"]},{"name":"Department of Central Laboratory, The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kun","family":"Yu","sequence":"additional","affiliation":[{"name":"Lianyungang Clinical College, Bengbu Medical University & The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]},{"name":"College of Life Sciences, Bengbu Medical University","place":["Bengbu, China"]},{"name":"Department of Central Laboratory, The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"JiaChen","family":"He","sequence":"additional","affiliation":[{"name":"Lianyungang Clinical College, Bengbu Medical University & The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]},{"name":"College of Life Sciences, Bengbu Medical University","place":["Bengbu, China"]},{"name":"Department of Central Laboratory, The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Wang","sequence":"additional","affiliation":[{"name":"Lianyungang Clinical College, Bengbu Medical University & The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]},{"name":"College of Life Sciences, Bengbu Medical University","place":["Bengbu, China"]},{"name":"Department of Central Laboratory, The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"WenJun","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Medicine Laboratory, The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"GuanHong","family":"Huang","sequence":"additional","affiliation":[{"name":"Lianyungang Clinical College, Bengbu Medical University & The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]},{"name":"Department of Central Laboratory, The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"XuZhu","family":"Gao","sequence":"additional","affiliation":[{"name":"Lianyungang Clinical College, Bengbu Medical University & The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]},{"name":"College of Life Sciences, Bengbu Medical University","place":["Bengbu, China"]},{"name":"Department of Central Laboratory, The Second People\u2019s Hospital of Lianyungang","place":["Lianyungang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,5,25]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"2876","DOI":"10.3390\/diagnostics14242876","article-title":"From tradition to innovation: diverse molecular techniques in the fight against infectious diseases","volume":"14","author":"Alsharksi","year":"2024","journal-title":"Diagn. 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