Differential Detection of the Tobamoviruses Tomato Mosaic Virus (ToMV) and Tomato Brown Rugose Fruit Virus (ToBRFV) Using CRISPR-Cas12a

CRISPR/Cas12a-based detection is a novel approach for the efficient, sequence-specific identification of viruses. Here we adopt use CRISPR/Cas12a to identify tomato brown rugose fruit virus (ToBRFV), new and emerging tobamovirus which causing substantial damage global industry. Specific CRISPR RNAs (crRNAs) were designed detect either ToBRFV or closely related mosaic (ToMV). This technology enabled differential ToMV. Sensitivity assays revealed that viruses can be detected from 15–30 ng RT-PCR product, specific could achieved mix ToMV ToBRFV. In addition, show this method enable in samples collected commercial greenhouses. These results demonstrate species-specific tobamoviruses. A future combination with isothermal amplification provide platform efficient user-friendly ways distinguish between strains resistance-breaking pathogens.