{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T14:52:15Z","timestamp":1784127135168,"version":"3.55.0"},"reference-count":31,"publisher":"PeerJ","license":[{"start":{"date-parts":[[2025,10,2]],"date-time":"2025-10-02T00:00:00Z","timestamp":1759363200000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The National Key R&D Program of China","award":["2023YFD1800404"],"award-info":[{"award-number":["2023YFD1800404"]}]},{"name":"The Science and Technology Program of Liaoning Province","award":["2024-MS-238"],"award-info":[{"award-number":["2024-MS-238"]}]},{"name":"The Project of the Department of Education of Liaoning Province","award":["LJ212410160093"],"award-info":[{"award-number":["LJ212410160093"]}]},{"name":"The Scientific Research Fund of the First Affiliated Hospital of Jinzhou Medical University","award":["KYTD-2022004"],"award-info":[{"award-number":["KYTD-2022004"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:p>Getah virus (GETV), a mosquito-borne alphavirus classified as a zoonotic disease, primarily infects livestock, particularly pigs and horses. In recent years, it has re-emerged in multiple Asian countries, posing a potential threat to animal husbandry and public health. In this study, we developed a rapid and sensitive GETV detection method based on reverse transcription-recombinase polymerase amplification (RT-RPA) and the clustered regularly interspaced short palindromic repeats (CRISPR)\/Cas12a system combined with a lateral flow dipstick (LFD) for visual readout. By leveraging sequence conservation in the GETV E2 envelope protein-coding regions, we engineered matched crRNA guides and amplification primers to develop a rapid CRISPR-Cas12a diagnostic workflow. The optimized platform combines RT-RPA (42\u00a0\u00b0C\/20 min) with Cas12a\u2019s trans-nuclease activity, permitting multiplex detection via real-time fluorescence quantification or immunochromatographic strip visualization. Analytical evaluation demonstrated a detection capability of 10 copies\/\u00b5L and exclusive specificity against four pathogen controls, including Japanese encephalitis virus and pseudorabies virus. Validation performed using simulated clinical samples revealed 100% concordance between the results of RT\u2013RPA\u2013CRISPR\/Cas12a\u2013LFD and quantitative polymerase chain reaction (PCR), while reducing the total detection time to 50 minutes. This approach eliminated the need for advanced instrumentation owing to its simplified operational design, enabling field-deployable rapid detection capabilities that establish essential technical infrastructure for initiating timely GETV containment measures. This approach has broad application potential in the fields of food safety, clinical diagnostics, and environmental science.<\/jats:p>","DOI":"10.7717\/peerj.20119","type":"journal-article","created":{"date-parts":[[2025,10,2]],"date-time":"2025-10-02T08:47:35Z","timestamp":1759394855000},"page":"e20119","source":"Crossref","is-referenced-by-count":4,"title":["Development and application of a CRISPR\/Cas12a-based reverse transcription\u2013recombinase polymerase amplification assay with lateral flow dipstick and fluorescence detection for Getah virus"],"prefix":"10.7717","volume":"13","author":[{"given":"Boyang","family":"Xia","sequence":"first","affiliation":[{"name":"Collaborative Innovation Center for Zoonosis Prevention and Treatment of Jinzhou Medical University,  Jinzhou, Liaoning, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyan","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Animal Husbandry and Veterinary Medicine, Jinzhou Medical University, Jinzhou, Liaoning,  China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiantian","family":"Fei","sequence":"additional","affiliation":[{"name":"Henan University of Chinese Medicine, Zhengzhou, Henan, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yueyu","family":"Ma","sequence":"additional","affiliation":[{"name":"Experimental Animal Center, Jinzhou Medical University, Jinzhou, Liaoning, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaxi","family":"Guo","sequence":"additional","affiliation":[{"name":"Experimental Animal Center, Jinzhou Medical University, Jinzhou, Liaoning, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dongliang","family":"Fei","sequence":"additional","affiliation":[{"name":"Experimental Animal Center, Jinzhou Medical University, Jinzhou, Liaoning, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiuwei","family":"Shu","sequence":"additional","affiliation":[{"name":"Liaoning Yikang Biological Co., Ltd, Liaoyang, Liaoning, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gang","family":"Zhao","sequence":"additional","affiliation":[{"name":"Liaoning Provinsiale Landbou-ontwikkelingsdienssentrum, Shenyang, Liaoning, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingxiao","family":"Ma","sequence":"additional","affiliation":[{"name":"College of Animal Husbandry and Veterinary Medicine, Jinzhou Medical University, Jinzhou, Liaoning,  China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongxia","family":"Yuan","sequence":"additional","affiliation":[{"name":"The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning, 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