{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T05:00:42Z","timestamp":1789621242268,"version":"build-2803163510"},"reference-count":63,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:00:00Z","timestamp":1764288000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:00:00Z","timestamp":1764288000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100005825","name":"National Institute of Food and Agriculture","doi-asserted-by":"publisher","award":["2023\u201067015\u201039481"],"award-info":[{"award-number":["2023\u201067015\u201039481"]}],"id":[{"id":"10.13039\/100005825","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Fish Diseases"],"published-print":{"date-parts":[[2026,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Tilapia Lake Virus (TiLV) is a significant threat to global tilapia aquaculture, highlighting the need for rapid and accurate diagnostic methods to manage outbreaks and minimise economic losses. This study presents the development and partial validation of a one\u2010pot assay integrating RT\u2010LAMP with the CRISPR\/Cas12b system for sensitive and specific TiLV detection. This assay amplifies viral RNA using RT\u2010LAMP, while CRISPR\/Cas12b enables a real\u2010time detectable signal. Targeting a conserved region in TiLV segment four, the assay achieves results within 75\u2009min at 62\u00b0C, with easy visualisation using a portable fluorescence viewer. It demonstrated high sensitivity, with a 95% limit of detection of 79.6 copies (95% CI: 48\u2013132 copies), and high specificity, with no cross\u2010reaction to other fish RNA or DNA viruses. Based on a validation panel of 261 samples from 9 source populations, the assay exhibited 92% diagnostic sensitivity (95% CI: 87%\u201396%) and 100% diagnostic specificity (95% CI: 97%\u2013100%). When assessed as a non\u2010lethal sample, gills provided a reliable and less invasive alternative despite lower viral loads compared to internal organs. Therefore, this partially validated one\u2010pot assay is potentially practical for enhancing TiLV detection and disease management in aquaculture systems, especially in field settings and resource\u2010limited laboratories.<\/jats:p>","DOI":"10.1111\/jfd.70087","type":"journal-article","created":{"date-parts":[[2025,11,29]],"date-time":"2025-11-29T05:59:36Z","timestamp":1764395976000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["One\u2010Pot\n                    <scp>RT<\/scp>\n                    \u2010\n                    <scp>LAMP CRISPR<\/scp>\n                    \/Cas12b Platform for Rapid Detection of Tilapia Lake Virus"],"prefix":"10.1111","volume":"49","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6160-2377","authenticated-orcid":false,"given":"Dorothea V.","family":"Megarani","sequence":"first","affiliation":[{"name":"Department of Infectious Diseases and Immunology, College of Veterinary Medicine University of Florida  Gainesville Florida USA"},{"name":"Emerging Pathogen Institute University of Florida  Gainesville Florida USA"},{"name":"Department of Clinical Pathology, Faculty of Veterinary Medicine Universitas Gadjah Mada  Yogyakarta Indonesia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5552-2920","authenticated-orcid":false,"given":"Lilia","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering, College of Engineering University of Florida  Gainesville Florida USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-8433-0422","authenticated-orcid":false,"given":"Hannah J.","family":"Siler","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases and Immunology, College of Veterinary Medicine University of Florida  Gainesville Florida USA"},{"name":"Emerging Pathogen Institute University of Florida  Gainesville Florida USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3723-0569","authenticated-orcid":false,"given":"Eva M.","family":"Quijano Card\u00e9","sequence":"additional","affiliation":[{"name":"Department of Medicine and Epidemiology, School of Veterinary Medicine University of California  Davis California USA"},{"name":"Animal Health Department Georgia Aquarium  Atlanta Georgia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0921-4796","authenticated-orcid":false,"given":"Christopher J.","family":"Martyniuk","sequence":"additional","affiliation":[{"name":"Department of Physiological Sciences, College of Veterinary Medicine University of Florida  Gainesville Florida USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6821-2232","authenticated-orcid":false,"given":"Paul M.","family":"Hick","sequence":"additional","affiliation":[{"name":"The Virology Laboratory Elizabeth Macarthur Agricultural Institute, NSW Department of Primary Industries and Regional Development  Menangle New South Wales Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4749-4004","authenticated-orcid":false,"given":"Joy A.","family":"Becker","sequence":"additional","affiliation":[{"name":"The University of Sydney, School of Life and Environmental Sciences  Camden New South Wales Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6054-9634","authenticated-orcid":false,"given":"Esteban","family":"Soto","sequence":"additional","affiliation":[{"name":"Department of Medicine and Epidemiology, School of Veterinary Medicine University of California  Davis California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5707-3476","authenticated-orcid":false,"given":"Win","family":"Surachetpong","sequence":"additional","affiliation":[{"name":"Institute of Aquaculture University of Stirling  Stirling UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4185-1867","authenticated-orcid":false,"given":"Roy P. 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