Sequences for detection of SARS-CoV-2 are provided and include example, SEQ ID NOs:4-8. The sequences are useful for amplifying the SARS-CoV-2 nsp8 gene in a sample, using Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP) and it is preferably used in combination with a Cas enzyme/sgRNA pair and a reporter nucleic acid.
The disclosed sequences can be use in methods of detecting SARS-CoV-2 nucleic acids in a sample. Generally, the specific gene sequence of SARS-CoV-2 RNA, herein nsp8, is amplified using RT-LAMP. The RT-LAMP products are scanned by the Cas12a-gRNA ribonucleoprotein (RNP) complex. The RNP binds to the specific complementary to gRNA, activating the transcleavage activity of Cas12a. The active Cas12a system cleaves a short ssDNA reporter that is labeled preferably, with a fluorophore and a quencher on either end. Cleavage of the reporter separates the quencher from the fluorophore, and fluorescence that is detectable with the naked eye is generated.