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

Area of Science:

• Veterinary Virology
• Molecular Diagnostics
• Public Health

Background:

• The zoonotic Getah virus (GETV) presents a growing risk to animal and human health.
• There is a critical need for rapid, sensitive, and field-deployable diagnostic tools for GETV surveillance.

Purpose of the Study:

• To develop and optimize a rapid, one-step, visual detection (ROSVD) platform for GETV.
• To create a diagnostic assay that is simple, fast, and does not require specialized equipment.

Main Methods:

• Integrated reverse transcription recombinase-aided amplification (RT-RAA) with CRISPR-EsCas13d-mediated collateral RNA cleavage for GETV detection.
• Employed a simplified sample preparation strategy involving rapid nucleic acid release, bypassing conventional extraction and purification.
• Completed the entire workflow, including amplification and detection, within 30 minutes at 37°C or ambient temperature (25°C).

Main Results:

• The ROSVD assay demonstrated sensitivity comparable to RT-qPCR at 37°C.
• Evaluated clinical specimens showed 100% concordance with RT-qPCR results.
• Detection results were visualized using ultraviolet light or a lateral flow assay.

Conclusions:

• The ROSVD platform is a rapid, sensitive, cost-effective, and instrument-independent diagnostic solution for GETV.
• This technology provides a practical approach for on-site GETV surveillance.
• The ROSVD platform serves as a versatile framework for detecting other emerging RNA pathogens.