Abstract: n this study, an RPA-CRISPR/Cas12a-LFS assay was developed for the detection of white spot syndrome virus （WSSV） based on recombinase polymerase amplification （RPA） combined with CRISPR/Cas12a system and lateral flow strips （LFS）, which was rapid, sensitive and simple to operate. The lower limit of the detection method for RPA-CRISPR/Cas12a-LFS was 1.12×101copies of vitiligo syndrome virus DNA per reaction, and the whole process could be completed within 1 h. Furthermore, the assay showed no cross-reactivity with other common shrimp viruses or healthy shrimp genomes. When testing 110 clinical shrimp samples collected from shrimp farms, the results of this study were completely consistent with the results of real-time polymerase chain reaction （PCR） assays. In conclusion, the detection method （RPA-CRISPR/Cas12a-LFS） constructed in this study shows great potential as a novel detection scheme for the early diagnosis of WSSV in shrimp.

Key words: white spot syndrome virus, recombinant polymerase amplification, CRISPR/Cas12a, lateral flow strip, virus detection