AML Mutation Analysis (ASXL1, CEBPA*, DNMT3A, FLT3*, IDH1, IDH2, KIT, NPM1, RUNX1, TET2, TP53, WT1) / Acute Myeloid Leukemia Mutational Analysis
• FLT3* and CEBPA* components performed via send-out testing and billed separately
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.
The AML gene analysis is a qualitative diagnostic test containing a subset of genes (ASXL1, DNMT3A, IDH1, IDH2, KIT, NPM1, RUNX1, TET2, TP53, WT1) combined with send out testing for FLT3 and CEPBA. Recent updates in NCCN guidelines and WHO classification indicate multiple additional genes are now recommended for testing in newly diagnosed AML patients, in addition to traditional cytogenetic analysis and FISH. The selected genetic markers in this analysis are associated with AML and identified as clinically significant by NCCN (National Comprehensive Cancer Network, 2018) and WHO guidelines (Swerdlow, S. et al., 2017). In addition to the risk stratification and prognostic information provided by identification of mutations in ASXL1, NPM1, RUNX and TP53, mutations in FLT3, IDH1/IDH2, and TP53 are associated with sensitivity to one or more drugs. Given the improved CMS reimbursement for 81450 (5-50 gene panel analysis for hematologic malignancies), targeted NGS panels provides 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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