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. 2020 Sep;22(9):1162–1178. doi: 10.1016/j.jmoldx.2020.06.006

Table 2.

AMP/TSCA Performance on 1° and 2° ITDs

MGH cohort (AMP) NGS ITDs in total Same size as 1° CE (total 1° CE ITDs) Same size as 2° CE (total 2° CE ITDs) Other size Unknown size
Custom FLT3-ITD 80 42 (42) 17 (17) 21 0
Clinical NGS calls 22 15 (42) 3 (17) 4 0
Novoalign unfiltered 39 23 (42) 8 (17) 8 0
BWH cohort (TSCA)
 Custom FLT3-ITD 72 48 (54) 17 (20) 3 4
 Clinical NGS calls 53 34 (54) 3 (20) 12 4

AML cohort (TSCA) Same size as 1° HC (total 1° HC ITDs) Same size as 2° HC (total 2° HC ITDs)
Custom FLT3-ITD 7 5 (5) 2 (2) 0 0
Clinical NGS calls 5 3 (5) 2 (2) 0 0

The sensitivity of the custom FLT3-ITD algorithm and clinical standard pipelines within clinical NGS panels (AMP and TSCA) to detect ITDs of the same size as CE or HC was the highest (100%) when applying the custom algorithm to the MGH AMP data. The ability to detect the primary ITD is particularly important because it contributes the most to an assessment of overall allelic ratio.

1°, Primary; 2°, secondary; AML, acute myeloid leukemia; AMP, anchored multiplex PCR; BWH, Brigham and Women's Hospital; CE, capillary electrophoresis; HC, hybrid-capture; ITD, internal tandem duplication; MGH, Massachusetts General Hospital; NGS, next-generation sequencing; TSCA, TruSeq Custom Amplicon.