CRISPR/Cas12a-based detection is a novel approach for the efficient, sequence-specific identification of viruses. In new research, CRISPR/Cas12a is used to identify the tomato brown rugose fruit virus (ToBRFV), a new and emerging tobamovirus that is causing substantial damage to the global tomato industry.
Specific CRISPR RNAs (crRNAs) were designed to detect either ToBRFV or the closely related tomato mosaic virus (ToMV). This technology enabled the differential detection of ToBRFV and ToMV.
Sensitivity assays revealed that viruses can be detected from 15–30 ng of RT-PCR product, and that specific detection could be achieved from a mix of ToMV and ToBRFV.
"In addition, we show that this method can enable the identification of ToBRFV in samples collected from commercial greenhouses. These results demonstrate a new method for species-specific detection of tobamoviruses," the researchers explain. "A future combination of this approach with isothermal amplification could provide a platform for efficient and user-friendly ways to distinguish between closely related strains and resistance-breaking pathogens."
Read the complete research here.
Alon, Dan & Hak, Hagit & Bornstein, Menachem & Pines, Gur & Spiegelman, Ziv. (2021). Differential Detection of the Tobamoviruses Tomato Mosaic Virus (ToMV) and Tomato Brown Rugose Fruit Virus (ToBRFV) Using CRISPR-Cas12a. Plants. 10. 1256. 10.3390/plants10061256.