graphical data file of Manuscript(Specific detection of RHDV2 by the RT-RPA-CRISPR/Cas14a1 system)

Rabbit hemorrhagic disease (RHD), also known as rabbit plague, is a peracute, highly contagious hemorrhagic disease caused by rabbit haemorrhagic disease virus (RHDV). recent years, the emerging variant RHDV2 has largely replaced the classical strains and become the dominant genotype, posing a serious threat to the rabbit industry. However,rapid and sensitive diagnostics methods suitable for field deployment remain limited. To address this need, we developed a nucleic-acid assay that combines reverse-transcription recombinase polymerase amplification (RT-RPA) with the CRISPR/Cas14a1 system for rapid detection of RHDV2. The assay operated at 32 °C - 46 °C and achieved a limit of detection of approximately 4.59 × 10¹ copies/μL. No cross-reactivity was observed with common rabbit pathogens, including rabbit rotavirus, Pasteurella multocida, Salmonella spp., Escherichia coli or rabbit coccidia. Results from clinical samples were fully concordant with RT-qPCR, indicating high specificity and sensitivity. This platform enables on-site rapid detection and epidemiological surveillance of RHDV2 and provides a new tool to support the prevention and control of this virus.