{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T20:19:51Z","timestamp":1790799591377,"version":"4.1.0"},"reference-count":44,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T00:00:00Z","timestamp":1768435200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T00:00:00Z","timestamp":1768435200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100003566","name":"Ministry of Oceans and Fisheries","doi-asserted-by":"publisher","award":["RS\u20102025\u201002373103"],"award-info":[{"award-number":["RS\u20102025\u201002373103"]}],"id":[{"id":"10.13039\/501100003566","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":[[2026,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Megalocytivirus pagrus 1<\/jats:italic>\n                    is a major viral pathogen that causes high mortality and economic losses in aquaculture worldwide. Although PCR\u2010based diagnostics are highly sensitive and specific, their equipment dependence and long assay times limit field applicability. In this study, a digital multienzyme isothermal rapid amplification with exonuclease probe (digital MIRA\u2010EXO) assay was developed and validated for rapid and specific detection of red sea bream iridovirus (RSIV), infectious spleen and kidney necrosis virus (ISKNV), and turbot reddish body iridovirus (TRBIV). The digital MIRA\u2010EXO assay specifically identified\n                    <jats:italic>Megalocytivirus pagrus 1<\/jats:italic>\n                    without cross\u2010reactivity with non\u2010target samples, including four fish cell lines and 13 different fish pathogens. Analytical sensitivity, expressed as 95% limit of detection, ranged from 146.87 to 201.6 copies\/\u03bcL across the three viruses. Diagnostic performance evaluation of 180 fish samples showed high sensitivity (92.22%), specificity (100%), and overall accuracy (96.11%), with an area under the receiver operating characteristic (ROC) curve of 0.971. Substantial agreement with reference assays was observed for experimentally infected (\u03ba\u2009=\u20090.786) and field samples (\u03ba\u2009=\u20090.792), and between column\u2010based and rapid DNA extraction methods (\u03ba\u2009=\u20090.783). Overall, the digital MIRA\u2010EXO assay provides a rapid, accurate, and field\u2010deployable diagnostic tool for\n                    <jats:italic>Megalocytivirus pagrus 1<\/jats:italic>\n                    detection.\n                  <\/jats:p>","DOI":"10.1111\/jfd.70125","type":"journal-article","created":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T10:12:14Z","timestamp":1768471934000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Development and Validation of a Multienzyme Isothermal Rapid Amplification\u2013Based Fluorescence Assay for Detection of\n                    <i>Megalocytivirus pagrus 1<\/i>"],"prefix":"10.1111","volume":"49","author":[{"given":"Eun Seo","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Aquatic Life Medicine Pukyong National University  Busan Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guk Hyun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Aquatic Life Medicine Pukyong National University  Busan Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7003-5696","authenticated-orcid":false,"given":"Kwang Il","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Aquatic Life Medicine Pukyong National University  Busan Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,1,15]]},"reference":[{"key":"e_1_2_9_2_1","first-page":"S49","article-title":"Limit of Blank, Limit of Detection and Limit of Quantitation","volume":"29","author":"Armbruster D. 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