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In this study, we integrated recombinase polymerase amplification (RPA) with CRISPR\/Cas12a-mediated detection to establish a fluorescence-based assay for rapid identification of P. plecoglossicida. The complete single-tube, two-stage RPA\u2013CRISPR\/Cas12a workflow can be performed within ~45 min. Using purified genomic DNA, the assay achieved an analytical detection limit of 1.65 copies \u03bcl\u22121 and showed no cross-reactivity with several other common fish pathogens. Its applicability was further evaluated using crude DNA extracts from spleen, liver, and kidney tissues of experimentally infected large yellow croakers. Overall, with its rapid turnaround, minimal equipment requirement, and high sensitivity, the RPA\u2013CRISPR\/Cas12a assay represents a promising diagnostic tool for rapid detection of P. plecoglossicida, thereby helping to control the spread of infection.<\/jats:p>","DOI":"10.1093\/femsle\/fnag067","type":"journal-article","created":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T11:35:24Z","timestamp":1781004924000},"source":"Crossref","is-referenced-by-count":0,"title":["The development of RPA\u2013CRISPR\/Cas12a assay for the detection of\n                    <i>Pseudomonas plecoglossicida<\/i>"],"prefix":"10.1093","volume":"373","author":[{"given":"Yiming","family":"Li","sequence":"first","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology , Shanghai 200237 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoran","family":"Du","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology , Shanghai 200237 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaxi","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology , Shanghai 200237 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingfan","family":"Xiao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology , Shanghai 200237 ,","place":["China"]},{"name":"Shanghai Engineering Research Center of Maricultured Animal Vaccines , Shanghai 200237 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Ma","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology , Shanghai 200237 ,","place":["China"]},{"name":"Shanghai Engineering Research Center of Maricultured Animal Vaccines , Shanghai 200237 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8706-0951","authenticated-orcid":false,"given":"Yibei","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology , Shanghai 200237 ,","place":["China"]},{"name":"Shanghai Engineering Research Center of Maricultured Animal Vaccines , Shanghai 200237 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7494-0869","authenticated-orcid":false,"given":"Qiyao","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering, Laboratory for Aquatic Animal Diseases, East China University of Science and Technology , Shanghai 200237 ,","place":["China"]},{"name":"Shanghai Engineering Research Center of Maricultured Animal Vaccines , Shanghai 200237 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,6,9]]},"reference":[{"key":"2026062512020828800_bib1","doi-asserted-by":"crossref","unstructured":"Ahmed MS, Jeba SM. \u00a0SalmonScan: a novel image dataset for machine learning and deep learning analysis in fish disease detection in aquaculture. 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