Spike-in validation of MRD assay
| Cell line DNA . | Expected VAF . | Total replicates, N . | Replicates ITD not detected . | Replicates ITD detected . | Sensitivity, % . |
|---|---|---|---|---|---|
| Cell line A (30-bp ITD) | 1E−02 | 4 | 0 | 4 | 100.00 |
| 1E−03 | 4 | 0 | 4 | 100.00 | |
| 1E−04 | 36 | 0 | 36 | 100.00 | |
| 5E−05 | 68 | 0 | 68 | 100.00 | |
| Cell line B (126-bp ITD) | 1E−02 | 4 | 0 | 4 | 100.00 |
| 1E−03 | 4 | 0 | 4 | 100.00 | |
| 1E−04 | 36 | 1 | 35 | 97.22 | |
| 5E−05 | 68 | 19 | 49 | 72.06 | |
| Jurkat cell line (WT) | 0 | 38 | 38 | 0 | N/A |
| Cell line DNA . | Expected VAF . | Total replicates, N . | Replicates ITD not detected . | Replicates ITD detected . | Sensitivity, % . |
|---|---|---|---|---|---|
| Cell line A (30-bp ITD) | 1E−02 | 4 | 0 | 4 | 100.00 |
| 1E−03 | 4 | 0 | 4 | 100.00 | |
| 1E−04 | 36 | 0 | 36 | 100.00 | |
| 5E−05 | 68 | 0 | 68 | 100.00 | |
| Cell line B (126-bp ITD) | 1E−02 | 4 | 0 | 4 | 100.00 |
| 1E−03 | 4 | 0 | 4 | 100.00 | |
| 1E−04 | 36 | 1 | 35 | 97.22 | |
| 5E−05 | 68 | 19 | 49 | 72.06 | |
| Jurkat cell line (WT) | 0 | 38 | 38 | 0 | N/A |
DNA from 2 cell lines with known FLT3-ITD mutations (cell line A: MV4-11, 30 bp homozygous; cell line B: PL-21, 126 bp heterozygous) was diluted into background DNA from an FLT3 wild-type cell line (Jurkat) at the ratios shown (“Expected”). The VAF is the mutation to total reads from the MRD assay and is shown for each cell line used as a source of mutant DNA (“Detected”).
N/A, not applicable; WT, wild-type.