{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T15:23:37Z","timestamp":1788276217138,"version":"build-2803163510"},"reference-count":40,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T00:00:00Z","timestamp":1776729600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T00:00:00Z","timestamp":1776729600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Fish Diseases"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Largemouth bass (\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Micropterus salmoides<\/jats:italic>\n                    <\/jats:styled-content>\n                    ) is an important freshwater\u2010cultured species in China.\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>M.\u2009salmoides<\/jats:italic>\n                    <\/jats:styled-content>\n                    rhabdovirus (MSRV) is a devastating pathogen that causes high mortality (exceeding 90% in acute outbreaks) in largemouth bass fry, posing a severe threat to the global largemouth bass aquaculture industry. To address the urgent demand for rapid, on\u2010site MSRV detection, two multienzyme isothermal rapid amplification (MIRA)\u2010derived methods\u2014multienzyme isothermal rapid amplification with fluorescence detection (MIRA\u2010FLU) and multienzyme isothermal rapid amplification with lateral flow dipstick (MIRA\u2010LFD)\u2014were established and optimized targeting the conserved nucleoprotein (N) gene of MSRV. The performance of both methods was systematically evaluated, and their practical applicability was verified using clinical samples collected from diseased fish farms. The results demonstrated that both methods exhibited high sensitivity, with a detection limit of 9 copies\/\u03bcL for the standard plasmid. Both detection methods demonstrated high specificity and showed no cross\u2010reactivity with other common aquatic viruses, indicating their high specificity for MSRV detection. MIRA\u2010FLU completed amplification within 25\u2009min at 42\u00b0C, while MIRA\u2010LFD achieved amplification within 12\u2009min at 37\u00b0C. MIRA\u2010FLU is used for real\u2010time fluorescent quantitative detection, whereas MIRA\u2010LFD enables visual, instrument\u2010free reading via lateral flow test strips. In comparative analyses of clinical samples, the diagnostic results of MIRA\u2010FLU and MIRA\u2010LFD were compared with those of the standard conventional PCR test and showed nearly 100% consistency. Therefore, our MIRA\u2010FLU and MIRA\u2010LFD detection methods demonstrate excellent specificity and sensitivity, providing two simple, rapid and reliable approaches for large\u2010scale field surveys of MSRV in resource\u2010limited settings.\n                  <\/jats:p>","DOI":"10.1111\/jfd.70191","type":"journal-article","created":{"date-parts":[[2026,4,22]],"date-time":"2026-04-22T04:29:56Z","timestamp":1776832196000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Development of Two Multienzyme Isothermal Rapid Amplification Techniques for the Rapid Visual Detection of\n                    <i>Micropterus salmoides<\/i>\n                    rhabdovirus"],"prefix":"10.1111","volume":"49","author":[{"given":"Qingyue","family":"Xu","sequence":"first","affiliation":[{"name":"School of Marine Science and Engineering Qingdao Agricultural University  Qingdao 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