Journal: BMC Medical Genomics
Article Title: Detection of leukemia gene fusions by targeted RNA-sequencing in routine diagnostics
doi: 10.1186/s12920-020-00739-4
Figure Lengend Snippet: A cryptic KMT2A -rearranged AML. The figure shows an AML with a KMT2A - MLLT4 gene fusion which is likely caused by an unbalanced translocation between chromosome 6 and 11. a . FISH-analysis using the KMT2A BA-probe ( KMT2A 5′ = green FISH-probe, KMT2A 3′ = red FISH-probe) could detect that a suspected KMT2A -rearrangement was present since deletion of the 3′-part (red) of the KMT2A -gene was seen. However, because of the lack of the reciprocal fusion, no fusion partner could be identified. b . The translocation was not visible with G-banding or FISH-analysis using KMT2A / MLLT4 dual fusion-probe ( KMT2A = red FISH-probe, MLLT4 = green FISH-probe). c . Archer anchored multiplex PCR and MiSeq sequencing revealed a KMT2A - MLLT4 exon 8-exon 2 fusion. The figure is a schematic overview of the sequences, a total of 924 reads spanning the breakpoint was scored. d . RT-PCR verified the KMT2A - MLLT4 gene fusion. P1 and P2 = patient 1 and 2 carrying KMT2A - MLLT4 e8-e2 gene fusions, P3 = patient 3 with a KMT2A - AFF1 gene fusion (negative control), NTC = non template control. For the original full length gel image see Supplementary Fig. 1
Article Snippet: Interphase FISH analysis was performed for screening of genomic aberrations depending on diagnosis and age at diagnosis using either an AML FISH probe panel (including probes for inv/t(16) Vysis LSI CBFB Break Apart (BA) rearrangement, t(15;17)(q22;q21) Vysis LSI PML/RARA Dual Color, Dual Fusion (DF) Translocation Probe kit, t(8;21)(q21;q22) Vysis LSI AML1/ETO Dual Color, DF Translocation Probe and 11q23-rearrangements Vysis LSI MLL Dual Color, BA Rearrangement probe, Abbott Laboratories, Chicago, Illinois) or the ALL FISH probe panel (including probes for del(9)(p21) Vysis LSI p16/CEP9, t(1;19)(q23;p13) Vysis LSI TCF3/PBX1 Dual Color, DF Translocation Probe, t(12;21)(p13;q22) Vysis LSI ETV6/RUNX1 Dual Color, DF Translocation Probe, t(9;22)(q34;q11) Vysis LSI BCR/ABL Dual Color, DF Translocation Probe, 11q23-rearrangements Vysis LSI MLL Dual Color, BA Rearrangement probe, Abbott Laboratories, Chicago, Illinois).
Techniques: Translocation Assay, Multiplex Assay, Sequencing, Reverse Transcription Polymerase Chain Reaction, Negative Control, Control