{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T07:27:29Z","timestamp":1791530849584,"version":"4.3.4"},"reference-count":52,"publisher":"SAGE Publications","issue":"2","license":[{"start":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T00:00:00Z","timestamp":1767744000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"funder":[{"DOI":"10.13039\/501100001503","name":"indian council of agricultural research","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001503","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Government of India"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["J VET Diagn Invest"],"published-print":{"date-parts":[[2026,3]]},"abstract":"<jats:p>\n                    White spot syndrome virus (WSSV; family\n                    <jats:italic toggle=\"yes\">Nimaviridae<\/jats:italic>\n                    ; taxon species\n                    <jats:italic toggle=\"yes\">White spot syndrome virus<\/jats:italic>\n                    ) is a major viral pathogen that poses a significant threat to the global shrimp industry, with early detection being the most effective strategy for disease control. We developed a CRISPR-Cas12a\u2013based dual-target detection assay for WSSV, specifically targeting the\n                    <jats:italic toggle=\"yes\">VP28<\/jats:italic>\n                    gene (gene product is a major envelope protein) and\n                    <jats:italic toggle=\"yes\">WSSV366<\/jats:italic>\n                    (a latency-associated gene), optimized using Indian WSSV isolates. Our CRISPR RNAs for both targets had high efficiency, and we evaluated the assay using fluorescence-based and lateral flow strip (LFS) endpoint detection. In fluorescence assays, the Cr-WSSV assay (without recombinase polymerase amplification, RPA) detected WSSV at 3\u2009\u00d7\u200910\u2075 copies\/\u03bcL; RPA integration significantly enhanced sensitivity, allowing detection at as low as 20 and 200 copies for\n                    <jats:italic toggle=\"yes\">VP28<\/jats:italic>\n                    and\n                    <jats:italic toggle=\"yes\">WSSV366<\/jats:italic>\n                    , respectively, with 100% specificity. We developed a CRISPR-based LFS assay with optimized FAM-biotin reporter concentrations of 100\u2009nM and 250\u2009nM, yielding robust and reproducible results for improved field applicability. Performance evaluation confirmed lack of cross-reactivity to other WOAH-listed shrimp pathogens, while maintaining detection limits of 20 and 200 copies of\n                    <jats:italic toggle=\"yes\">VP28<\/jats:italic>\n                    and\n                    <jats:italic toggle=\"yes\">WSSV366<\/jats:italic>\n                    . Clinical validation further demonstrated that the RPA-Cr-WSSV-LFS assay successfully detected\n                    <jats:italic toggle=\"yes\">WSSV366<\/jats:italic>\n                    even in\n                    <jats:italic toggle=\"yes\">VP28<\/jats:italic>\n                    -negative samples, underscoring the importance of detecting\n                    <jats:italic toggle=\"yes\">WSSV366<\/jats:italic>\n                    in latent infections. Our rapid, cost-effective, and highly sensitive CRISPR-Cas\u2013based assay enhances WSSV surveillance and biosecurity in shrimp aquaculture by incorporating structural and latency-associated gene markers, making it a promising alternative to conventional molecular testing.\n                  <\/jats:p>","DOI":"10.1177\/10406387251399147","type":"journal-article","created":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T07:04:08Z","timestamp":1767855848000},"page":"192-204","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":2,"title":["CRISPR-Cas12a\u2013based lateral flow detection of white spot syndrome virus: a dual-target approach for detection of early and latent infection"],"prefix":"10.1177","volume":"38","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-0865-4332","authenticated-orcid":false,"given":"Chandraprakasham","family":"Manojkumar","sequence":"first","affiliation":[{"name":"University of Tasmania, Hobart, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-0955-9963","authenticated-orcid":false,"given":"Maharshi","family":"Limbola","sequence":"additional","affiliation":[{"name":"ICAR\u2013Central Institute of Fisheries Education, Mumbai, Maharashtra, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samrat","family":"Paul","sequence":"additional","affiliation":[{"name":"ICAR\u2013Indian Agricultural Research Institute, New Delhi, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-5728-3769","authenticated-orcid":false,"given":"Kaliyamoorthy","family":"Thangadurai","sequence":"additional","affiliation":[{"name":"ICAR\u2013Central Institute of Fisheries Education, Mumbai, Maharashtra, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kooloth Valappil","family":"Rajendran","sequence":"additional","affiliation":[{"name":"Central University of Kerala, Kasaragod, Kerala, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anirban","family":"Roy","sequence":"additional","affiliation":[{"name":"ICAR\u2013Indian Agricultural Research Institute, New Delhi, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bikash","family":"Mandal","sequence":"additional","affiliation":[{"name":"ICAR\u2013Indian Agricultural Research Institute, New Delhi, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kezhedath","family":"Jeena","sequence":"additional","affiliation":[{"name":"ICAR\u2013Central Institute of Fisheries Education, Mumbai, Maharashtra, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3558-7529","authenticated-orcid":false,"given":"Megha Kadam","family":"Bedekar","sequence":"additional","affiliation":[{"name":"ICAR\u2013Central Institute of Fisheries Education, Mumbai, Maharashtra, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2026,1,7]]},"reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.aquaculture.2019.734340"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aar6245"},{"key":"e_1_3_3_4_2","first-page":"101016","article-title":"Probiotics expressing double-stranded RNA targeting VP28 efficiently protect shrimps from WSSV infection","volume":"23","author":"Dekham K","year":"2022","unstructured":"Dekham K, et al. Probiotics expressing double-stranded RNA targeting VP28 efficiently protect shrimps from WSSV infection. Aquac Rep 2022;23:101016.","journal-title":"Aquac Rep"},{"key":"e_1_3_3_5_2","unstructured":"Food and Agriculture Organization (FAO). The State of World Fisheries and Aquaculture. Blue Transformation in Action. 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CRISPR-Cas fluorescent cleavage assay coupled with recombinase polymerase amplification for sensitive and specific detection of Enterocytozoon hepatopenaei. Biotechnol Rep (Amst) 2020;27:e00485.","journal-title":"Biotechnol Rep (Amst)"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1111\/jfd.14059"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.18.10162-10167.2003"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.aquaculture.2023.739968"},{"key":"e_1_3_3_17_2","article-title":"Diagnostic validity of molecular diagnostic assays for white spot syndrome virus at different severity grades","volume":"9","author":"Kim MJ","year":"2023","unstructured":"Kim MJ, et al. Diagnostic validity of molecular diagnostic assays for white spot syndrome virus at different severity grades. 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