{"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":1789621242255,"version":"build-2803163510"},"reference-count":37,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T00:00:00Z","timestamp":1764633600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T00:00:00Z","timestamp":1764633600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32202994"],"award-info":[{"award-number":["32202994"]}],"id":[{"id":"10.13039\/501100001809","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>\n                    Tilapia parvovirus (TiPV) is an emerging pathogen associated with high mortality rates in farmed tilapia, highlighting the urgent need for rapid and accurate diagnostic tools. In this study, we established an RPA\u2010CRISPR\/Cas12a detection system targeting the TiPV NS1 gene. The assay conditions were systematically optimised, including 15\u2010min RPA amplification at 39\u00b0C, with reagent concentrations of 200\u2009nM Cas12a, 250\u2009nM crRNA and 200\u2009nM ssDNA reporter. Specificity tests showed no cross\u2010reactivity with other tilapia pathogens (TiLV,\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>S. agalactiae<\/jats:italic>\n                    <\/jats:styled-content>\n                    ) and other aquatic pathogens (LMBRaV, YcCV, GCRV II, WSSV, CyHV\u20102, SVCV). Sensitivity evaluation revealed a limit of detection (LoD) of 1.97\u2009\u00d7\u200910\n                    <jats:sup>1<\/jats:sup>\n                    copies\/\u03bcL, which was 100\u2010fold more sensitive than PCR (1.97\u2009\u00d7\u200910\n                    <jats:sup>3<\/jats:sup>\n                    copies\/\u03bcL). Clinical validation with 20 tilapia samples demonstrated a 50% positive detection rate for RPA\u2010CRISPR\/Cas12a, 15% higher than PCR (35%). This integrated method combines the advantages of RPA and CRISPR\u2010based signal transduction, offering a field\u2010applicable solution for TiPV monitoring in resource\u2010limited aquaculture environments.\n                  <\/jats:p>","DOI":"10.1111\/jfd.70095","type":"journal-article","created":{"date-parts":[[2025,12,2]],"date-time":"2025-12-02T10:30:10Z","timestamp":1764671410000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Development of an\n                    <scp>RPA<\/scp>\n                    \u2010\n                    <scp>CRISPR<\/scp>\n                    \u2010Cas12a Fluorescence Assay for Rapid and Sensitive Detection of Tilapia Parvovirus (\n                    <scp>TiPV<\/scp>\n                    )"],"prefix":"10.1111","volume":"49","author":[{"given":"Chu","family":"Ma","sequence":"first","affiliation":[{"name":"Yangtze River Fisheries Research Institute, Chinese Academy of Fishery 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