Linearity and validation of the IGHV leader-based NGS assay. Comparisons between expected and observed MRD measurements, or cross-technique MRD comparisons. Each dot represents 1 measurement. All Pearson correlation coefficients were computed using the log10-transformed values. (A) Comparison between expected and observed MRD measurements obtained by measuring MRD on contrived samples using the IGHV leader-based NGS assay. The notch represents the median, the upper and lower hinges signify the first and third quartiles, respectively, and the whiskers extend to the largest value within 1.5 times the interquartile range. The value above the boxplots denotes the amount of input DNA used for this MRD depth. The black line indicates a linear regression between expected and observed values; the shaded area indicates the 95% confidence interval for the regression mean. The red dotted lines indicate a threefold difference between expected and observed data points. Data points have horizontal jitter to improve their visibility. (B) Comparison between measurements on contrived samples obtained by ASO-qPCR and IGHV leader-based NGS. The black dotted line represents a perfect linear relationship. The line between data points signifies the range of the replicates per rearrangement. (C) Comparison between expected and observed MRD measurements obtained by measuring MRD on contrived samples using ddPCR. The black line indicates a linear regression between expected and observed values; the shaded area indicates the 95% confidence interval for the regression mean. The red dotted lines indicate a threefold difference between expected and observed data points. (D) Comparison between measurements on contrived samples obtained by ASO-qPCR and IGHV leader-based NGS. The black dotted line represents a perfect linear relationship. The line between data points signifies the range of the replicates per rearrangement. (E) Comparison between measurements on contrived samples obtained by ASO-qPCR and IGHV leader-based NGS. The black dotted line represents a perfect linear relationship. The red dotted lines indicate a threefold difference between expected and observed data points. The value above the measurements denotes the amount of input DNA used for this MRD depth. ID, identifier.
