A Rapid, Field-Deployable Diagnostic Platform for Getah Virus Based on RT-RAA and CRISPR EsCas13d

The emerging zoonotic Getah virus (GETV) poses an increasing threat to both animal and human health, underscoring the need for rapid, sensitive and field-deployable diagnostic tools. In this study, we developed and optimized a rapid, one-step, visual detection (ROSVD) platform for GETV by integrating reverse transcription recombinase-aided amplification (RT-RAA) with CRISPR-EsCas13d-mediated collateral RNA cleavage. Notably, the ROSVD assay uses a simplified sample-preparation strategy based on rapid nucleic acid release, eliminating the need for conventional nucleic acid extraction and purification. The entire workflow, including amplification and detection, is completed within 30 min at 37°C or ambient temperature (25°C) without specialized instrumentation. Detection results can be visualized directly under ultraviolet light or with a lateral flow assay. At 37°C, the ROSVD assay achieved sensitivity comparable to RT-qPCR, and evaluation of clinical specimens showed 100% concordance with RT-qPCR results. Collectively, these findings demonstrate that ROSVD is a rapid, sensitive, cost-effective and instrument-independent diagnostic platform, providing a practical solution for on-site surveillance of GETV and a versatile framework for the detection of other emerging RNA pathogens.