AML Mutation Panel (ASXL1, CEBPA*, DNMT3A, FLT3*, IDH1, IDH2, KIT, NPM1, RUNX1, TET2, TP53, WT1) / Acute Myeloid Leukemia Mutational Analysis Panel by Next Generation Sequencing, FLT3 and CEBPA by PCR and PCR/Sequencing, respectively.
Acute Myeloid Leukemia (AML)
- AML mutational testing will be performed simultaneously with cytogenetic testing.
- FLT3 testing by PCR (send-out assay) will be initiated in parallel with AML mutation testing by NGS.
- CEBPA* testing will be performed as a reflex send out test, if both AML mutation testing by NGS and FLT3 by PCR testing are negative.
Next Generation Sequencing on the Illumina MiSeqDx
PCR and PCR/Sequencing for FLT3 and CEBPA testing, respectively
5.0 mL (min 3.0 mL) peripheral blood or (min 2.0 mL) bone marrow in EDTA, Sodium Heparin or ACD
18-25°C for 1 week; 2 -8°C for up to 14 days
Storage & Handling
Ship at ambient temperature, cool in summer. (Do not allow cool/refrigerated pack to directly contact sample.)
Causes for Rejection
Improper specimen labeling; Insufficient sample volume; Clotted specimen; Specimen older than 7 days
SNVs and small to mid-size insertion/deletions (<100 bp mutations) in the following genes: ASXL1, CEBPA, DNMT3A, FLT3, IDH1, IDH2, KIT, NPM1, RUNX1, TET2, TP53, WT1. The assay does not detect whole gene or exon insertion and deletions, copy number variants or gene rearrangements (fusions). The potential for false negatives for genetic mutations increases in samples with <10% disease type cellularity.
Genetic testing in AML has traditionally included classical cytogenetic analysis and FISH. The role of molecular testing has been primarily limited to risk stratification in normal karyotype AMLs (FLT3, NPM1, CEBPa); KIT mutation testing has also been performed to further refine prognosis among core binding factor leukemias - AML with t(8;21), and AML with inv(16). Given the recent updates in NCCN guidelines and WHO recommendations, multiple additional genes are now recommended / strongly recommended for testing in AML. Given also the improved CMS reimbursement for 81450 (5-50 gene panel analysis for hematologic malignancies), targeted NGS panels provide a practical and feasible solution for optimal molecular genetic characterization of AML at initial diagnosis in the routine clinical laboratory setting.
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