Eurofins Biomnis code

MYSLA

Specialty

Genetics

Clinical significance

The ¿AML¿ NGS panel includes the analysis of 41 genes: ASXL1/BCOR/BCORL1/BRAF/CALR/CBL/CEBPA/CSF3R/DNMT3A/ETNK1/ETV6/EZH2/FLT3/GATA2/GNB1/HRAS/IDH1/IDH2/JAK2/KIT/KMT2A-MLL/KRAS/MPL/NF1/NPM1/NRAS/PHF6/PPM1D/PRPF8/PTPN11/RUNX1/SETBP1/SF3B1/SRSF2/STAG2/TET2/TP53/UBA1/U2AF1/WT1/ZRSR2. It can be used in the three areas of diagnosis, prognosis and theranostics and must be associated with a cytogenetic study to classify an AML and define its prognosis. - Two classifications exist for an AML diagnosis with mutational data: the WHO 2022 classification and the ICC/ELN 2022 classification. * WHO 2022: the NPM1 and CEBPA mutational status is used to define 2 entities of "AML with defining genetic abnormalities". Rq : for CEBPA, the concepts of biallelic mutations (whatever the type of mutation) (biCEBPA) or a monoallelic mutation of the bZIP domain (smbZIP-CEBPA) are retained. * ICC/ELN 2022: the NPM1, CEBPA and TP53 mutational status is used to define 3 entities of "AML with defining genetic abnormalities". Rq : for CEBPA, it is exclusively a bZIP domain mutation (monoallelic or biallelic). For both classifications, in the context of "AML with defining genetic abnormalities", 10% of blasts are sufficient (and no longer 20%) to make the diagnosis of AML (exception: for WHO 2022: 20% for CEBPA and BCR::ABL1 and whatever the percentage of blasts for NPM1 - for ICC/ELN 2022: 20% for TP53 and BCR::ABL1). * For the entity "AML, myelodysplasia-related¿, in addition to the search for defined cytogenetic abnormalities, analysis of mutations in the following 8 genes must be performed according to WHO 2022: ASXL1, BCOR, EZH2, STAG2, SF3B1, SRSF2, U2AF1 and ZRSR2. The ICC/ELN 2022 also includes a 9th gene: RUNX1. - The AML NGS panel also provides prognostic support (ICC/ELN 2022 recommendations) by allowing the definition of molecular factors for a favorable, intermediate or unfavorable prognosis: * Favorable : - NMP1 mutation without FLT3-ITD mutation - monoallelic or biallelic mutation of the bZIP domain of CEBPA * Intermediate : - FLT3-ITD mutation (irrespective of allelic ratio) with or without NPM1 mutation. * Unfavorable : - TP53 mutation (VAF of at least 10%), frequently associated with a complex karyotype. - ASXL1, BCOR, EZH2, RUNX1, SF3B1, SRSF2, STAG2, U2AF1 or ZRSR2 mutations. These markers should not be used as an adverse prognostic marker if they co-occur with favorable-risk AML subtypes. Rq : FLT3-ITD/FLT3wt ratio is no longer used. - In theranostic, testing for FLT3 mutation is already a prerequisite for a targeted treatment. IDH1, IDH2 and TP53 mutations may also represent therapeutic targets.

Preanalytics

• 2 ml EDTA bone marrow or 2 x 5 ml EDTA whole blood if blast infiltration into the periphery is significant or DNA extracted : (200ng of DNA minimum)
•   Ambient temperature

• A tube specifically for this analysis : No

Further information

Samples to be collected from Monday to Friday
Sample must be refrigerated if transport is > 48 hours
Use the specific request form B8-INTGB: Malignant blood disorders
Attach:
- the results of the FBC-platelets
- the myelogram report
- the immunophenotyping report

Specific equipment available

• S9L: Envelopes yellow for karyotypes LYON

Documents to download

• Specific request form B8-INTGB
• Panel form (DS69-INTGB-AML)
• Information aimed at healthcare professionals(DS3-INTGB)

Methodology

Method: Targeted sequencing of genes (NGS). - Library: Library Preparation Enzymatic Fragmentation Kit 2.0 Twist - NGS Platform: NovaSeq Illumina (2x150pb) - paired-end sequencing - Analysis software: Demultiplexage BCLconvert V3.10.5 / VarSome Pipeline version 11.8 (CE-IVD)

Turnaround time

10 days (Results may require an extended turnaround time, one week, depending on the confirmation tests required by Sanger sequencing)