Amplification- free detection of zoonotic viruses using Cas13 and multiple CRISPR RNAs

This study developed an amplification-free CRISPR-Cas13 assay for detecting hantavirus and influenza A virus RNA. Cas13 was used to directly detect viral RNA without PCR amplification. Multiple crRNAs were designed to improve detection sensitivity. Both hantaviruses and influenza A virus were successfully detected. In hantaviruses, the use of multiple crRNAs increased detection sensitivity, whereas no significant improvement was observed for influenza A virus. The assay could detect viral RNA from infected cultured tissues and lung samples without amplification. In clinical samples, the positivity rate showed 85% agreement with RT-qPCR results, suggesting the potential utility of this method for rapid viral diagnostics and surveillance.
(AI)