Fig. 2

Combined analysis of variants with the DNA Amplicon and Pindel apps to evaluate the performance of the custom 22-gene NGS panel based on two identical but independent NGS runs using reference standards. The MPL W515L variant in the Seraseq Myeloid Mutation Mix (Seraseq) was not detected in one of the replicates, giving the custom NGS panel a sensitivity of 99.2%. One variant was detected in the wild-type reference standard TruQ0, giving the panel a specificity of 96.3%. The custom NGS panel also had a positive predictive value of 97.7%, an average intra-run and inter-run concordance of 98.8% [range 95.2–100%] and 99.0% [range 95.2–100%] respectively, and was able to detect variants at as low as 1% allele frequency. FP, False positive; TP, True positive; FN; False negative; Rep 1, Replicate 1; Rep 2, Replicate 2