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Abstract
The limit of detection for the RPA-CRISPR/Cas12a detection system targeting the TiPV NS1 gene is 1.97 × 10 copies/μL, making it 100-fold more sensitive than traditional PCR.
- The RPA-CRISPR/Cas12a system showed a 50% positive detection rate in clinical validation with 20 tilapia samples.
- This detection rate was 15% higher than that achieved with PCR, which had a 35% positive detection rate.
- Specificity tests confirmed no cross-reactivity with other tilapia and aquatic pathogens.
- The assay was optimized for 15-minute amplification at 39°C using specific reagent concentrations.
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