Detection of the SARS‐CoV‐2 D614G mutation using engineered Cas12a guide RNA

Detection of pathogens with single-nucleotide variations is indispensable for the disease tracing, but remains technically challenging. The D614G mutation in SARS-CoV-2 spike protein known to markedly enhance viral infectivity difficult detect. Here, we report an effective approach called “synthetic mismatch integrated crRNA guided Cas12a detection” (symRNA-Cas12a) detect D614 and G614 variants effectively. Using this method, systemically screened a pool crRNAs that contain all possible nucleotide substitutions covering -2 +2 positions around identify one can efficiently increase detection specificity by 13-fold over ancestral crRNA. With selected crRNA, symRNA-Cas12a assay as low 10 copies synthetic mutant RNA results are confirmed be accurate Sanger sequencing. Overall, have developed method specifically, sensitively rapidly mutation.