Fig. 2

EasyCatch is applicable to clinical samples and other cancer mutations. a Schematic diagram of the whole mutation diagnosis. b EasyCatch and FGS results of 32 AML samples with known FLT3-D835 mutation status, 10/32 cases are shown. D835Y/V/H/F-positive patients are marked by red IDs, and red triangles indicate mutant bases. c EasyCatch, FGS, and NGS results of 80 AML samples with unknown FLT3-D835 mutation status, 9/80 cases are shown. WT and mutated bases in NGS are marked by green and red, respectively. d Statistical table of the sensitivity and specificity of EasyCatch compared with FGS using NGS as a standard reference. e Sensitivity comparison between EasyCatch and CRISPR detection in the detection of IDH2-R172K, EGFR-e19del and L858R, and NRAS-G12D mutations. Genomic locations of these mutations were shown above, wherein exons and mutation sites were colored in blue and red, respectively. The tested samples were 1e5 copies of plasmid templates with a mutation rate of 0.1%. Each amplified product was detected by both WT-crRNA and mutation-crRNA induced Cas12a reaction. Fluorescence intensity and naked eye results were both recorded. f Statistic analysis of the MT/WT fluorescence ratio in EasyCatch and CRISPR detection. The results of 1 and 0.1% mutated samples were counted together. g, h, i The qPCR assay for EGFR-e19del, L858R and NRAS-G12D detection, respectively. The qPCRs were performed on 10, 1 and 0.1% mutated templates. A 100% WT template and ddH₂O (NC) served as control. j The statistical chart of restriction enzyme cuttable human disease-related genomic sites (mutation < 27 bp, which is the detection length of crRNA), wherein commercial available 37 °C restriction enzyme cuttable sites can be potentially detected by EasyCatch. The inclusion relation is shown in the upper-right corner