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Current diagnostic methods, such as PCR, are limited by equipment dependency and procedural complexity, hindering point-of-care (POC) applications. To address this, we developed an integrated assay combining recombinase-aided amplification (RAA) with CRISPR-Cas13a-mediated SHERLOCK technology and lateral flow detection (LFD) for rapid and visual detection of CyHV-3 in clinical samples. The KHV-SHERLOCK assay targets a conserved region of the CyHV-3 thymidine kinase (TK) gene, demonstrating exceptional specificity with no cross-reactivity to related pathogens or host DNA. Sensitivity evaluations revealed a detection limit of 100 ag\/μL for CyHV-3 plasmid DNA, tenfold more sensitive than the conventional PCR (1 fg\/μL) assay, even in the presence of 100 ng of carp genomic DNA as background interference. Clinical validation using 50 archived samples showed 100% concordance with reference PCR results, confirming diagnostic reliability. The assay's isothermal RAA step (37°C, 40 min) and CRISPR-Cas13a detection (37°C, 1 h) enable equipment-free operation, while LFD provides unambiguous visual results within minutes. This platform merges high sensitivity with POC practicality, offering a transformative tool for field-based KHVD surveillance."}]}],"bf:provisionActivity":[{"@type":"bf:Publication","bf:agent":[{"@type":"schema:Organization","schema:name":[{"@value":"Wiley"}]}]}],"dce:rights":[{"@language":"eng","@value":"© 2025 The Author(s)"}],"dce:subject":[{"@type":"skos:Concept","skos:prefLabel":[{"@language":"eng","@value":"CRISPR-Cas13a"}]},{"@type":"skos:Concept","skos:prefLabel":[{"@language":"eng","@value":"CyHV-3"}]},{"@type":"skos:Concept","skos:prefLabel":[{"@language":"eng","@value":"Lateral Flow Detection"}]},{"@type":"skos:Concept","skos:prefLabel":[{"@language":"eng","@value":"Point-of-care Diagnostics"}]},{"@type":"skos:Concept","skos:prefLabel":[{"@language":"eng","@value":"Recombinase-aided Amplification"}]},{"@type":"skos:Concept","skos:prefLabel":[{"@language":"eng","@value":"SHERLOCK"}]}],"dce:title":[{"@type":"bf:Title","bf:mainTitle":[{"@language":"eng","@value":"CRISPR‐Enhanced RAA‐SHERLOCK Assay for Point‐of‐Care Detection of Cyprinid Herpesvirus‐3: Development, Validation and Clinical Application"}]}],"dcterms:accessRights":[{"@type":"skos:Concept","skos:exactMatch":["https:\/\/pid.phaidra.org\/vocabulary\/QW5R-NG4J"],"skos:prefLabel":[{"@language":"eng","@value":"open access"},{"@language":"ita","@value":"Open Access"}]}],"dcterms:issued":["2026"],"dcterms:language":["eng"],"dcterms:type":[{"@type":"skos:Concept","skos:exactMatch":["https:\/\/pid.phaidra.org\/vocabulary\/69ZZ-2KGX"],"skos:prefLabel":[{"@language":"eng","@value":"Text"},{"@language":"deu","@value":"Text"},{"@language":"ita","@value":"Documento PDF"}]}],"ebucore:filename":["Journal of Fish Diseases - 2025 - Soliman - CRISPR‐Enhanced RAA‐SHERLOCK Assay for Point‐of‐Care Detection of Cyprinid.pdf"],"ebucore:hasMimeType":["application\/pdf"],"edm:hasType":[{"@type":"skos:Concept","skos:exactMatch":["https:\/\/pid.phaidra.org\/vocabulary\/VKA6-9XTY"],"skos:prefLabel":[{"@language":"eng","@value":"journal article"},{"@language":"deu","@value":"Wissenschaftlicher Artikel"},{"@language":"ita","@value":"Articolo scientifico"}]}],"edm:rights":["http:\/\/creativecommons.org\/licenses\/by\/4.0\/"],"oaire:version":[{"@type":"skos:Concept","skos:exactMatch":["https:\/\/pid.phaidra.org\/vocabulary\/PMR8-3C8D"],"skos:prefLabel":[{"@language":"eng","@value":"version of record (published version)"},{"@language":"ita","@value":"Versione editoriale\/Version of Record"}]}],"rdam:P30004":[{"@type":"ids:doi","@value":"10.1111\/jfd.70079"}],"rdau:P60193":[{"@type":"schema:CreativeWork","bibo:issue":[{"@value":"4"}],"bibo:volume":[{"@value":"49"}],"dce:title":[{"@type":"bf:Title","bf:mainTitle":[{"@language":"eng","@value":"Journal of Fish Diseases"}]}],"dcterms:issued":["2026"],"ids:issn":["0140-7775"]}],"role:aut":[{"@type":"schema:Person","schema:affiliation":[{"@type":"schema:Organization","schema:name":[{"@value":"University of Veterinary Medicine Vienna"}],"skos:exactMatch":["https:\/\/ror.org\/01w6qp003"]}],"schema:familyName":[{"@value":"Soliman"}],"schema:givenName":[{"@value":"Hatem"}],"skos:exactMatch":[{"@type":"ids:orcid","@value":"0000-0003-1641-6759"}]},{"@type":"schema:Person","schema:affiliation":[{"@type":"schema:Organization","schema:name":[{"@value":"University of Veterinary Medicine Vienna"}],"skos:exactMatch":["https:\/\/ror.org\/01w6qp003"]}],"schema:familyName":[{"@value":"Akram"}],"schema:givenName":[{"@value":"Naveed"}]},{"@type":"schema:Person","schema:affiliation":[{"@type":"schema:Organization","schema:name":[{"@value":"University of Veterinary Medicine Vienna"}],"skos:exactMatch":["https:\/\/ror.org\/01w6qp003"]}],"schema:familyName":[{"@value":"Saleh"}],"schema:givenName":[{"@value":"Mona"}],"skos:exactMatch":[{"@type":"ids:orcid","@value":"0000-0001-9273-1502"}]}],"schema:numberOfPages":["8"]}