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<lom:langstring xml:lang="x-none">10.1111/jfd.70079</lom:langstring>

  
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<lom:langstring xml:lang="en">CRISPR‐Enhanced RAA‐SHERLOCK Assay for Point‐of‐Care Detection of Cyprinid Herpesvirus‐3: Development, Validation and Clinical Application</lom:langstring>

  
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<lom:langstring xml:lang="en">Koi herpesvirus disease (KHVD), caused by Cyprinid herpesvirus-3 (CyHV-3), poses a significant threat to global aquaculture due to its high mortality rates and economic impact. 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&#39;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.</lom:langstring>

  
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<lom:language>eng</lom:language>

  
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<lom:langstring xml:lang="en">CRISPR-Cas13a</lom:langstring>

  
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<lom:langstring xml:lang="en">CyHV-3</lom:langstring>

  
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<lom:langstring xml:lang="en">Lateral Flow Detection</lom:langstring>

  
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<lom:keyword>
  
<lom:langstring xml:lang="en">Point-of-care Diagnostics</lom:langstring>

  
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<lom:keyword>
  
<lom:langstring xml:lang="en">Recombinase-aided Amplification</lom:langstring>

  
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<lom:langstring xml:lang="en">SHERLOCK</lom:langstring>

  
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<lom:datetime>2026-08-05T08:43:55.001Z</lom:datetime>

  
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<lom:vcard>BEGIN:VCARD
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N:Soliman;Hatem;
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N:Akram;Naveed;
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<lom:vcard>BEGIN:VCARD
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N:Saleh;Mona;
FN:Mona Saleh
X-ORCID:https://orcid.org/0000-0001-9273-1502
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