Fig. 3: Clonality assessment of classic Hodgkin lymphoma by next-generation sequencing as compared to BIOMED-2/GeneScan assay.

The results of clonality analysis for (A) fresh frozen (FF) and (B) formalin-fixed paraffin-embedded (FFPE) tissue of case 12 are shown for all IG targets (IGHV-IGHD-IGHJ, IGHD-IGHJ, IGKV-IGKJ, and IGKV/Intron RSS-KDE) based on next-generation sequencing of immunoglobulin gene rearrangements (IG-NGS) (left panels) and BIOMED-2/GeneScan assay (right panels). IG-NGS clonality results: junction length in amino acids (junction aa length) is shown on the x-axis, the abundancy of clonotypes is shown in percentages on the y-axis. Each bar represents a total of all detected clonotypes with the same junction length, each color represents a unique clonotype based on their nucleotide sequence. The 50 most-abundant clonotypes are shown in colors, all other clonotypes are depicted together in gray. Clonotypes are described by the 5′ and 3′ gene annotation and the junctional nucleotide sequence of the rearrangement. BIOMED-2/GeneScan clonality results: clonal product is marked with black arrow for IGKV/Intron RSS-KDE in FFPE sample; ^*non-specific IGHD-IGHJ non-specific germline product; ^#IGKV-IGKJ polyclonal signal, the narrow Gaussian curve in the IGKV 1f/6/7-IGKJ size window is due the high homology of the IGKV genes and explains why minor clonal rearrangements can be easily missed.