Eurofins Biomnis code

MYSMO

Specialty

Genetics

Clinical significance

The "CMML" NGS panel includes the analysis of 23 genes: ASXL1/BCOR/CALR/CBL/DNMT3A/EZH2/FLT3/IDH1/IDH2/JAK2/KRAS/MPL/NF1/NPM1/NRAS/RUNX1/SETBP1/SF3B1/SRSF2/TET2/TP53/U2AF1/ZRSR2. It can be used in the three areas of diagnosis, prognosis and theranostics. It must be associated with a bone marrow cytogenetic analysis. - The panel is most widely appreciated for it prognostic value, as it is used to calculate the CMML CPSS-Mol prognostic score. This score is calculated from clinico-biological data, i.e., transfusion requirements, CBC-platelet data (WBC count), percentage of bone marrow blasts. It also includes cytogenetic data and the presence or absence of mutations in the ASXL1, NRAS, RUNX1 and SETBP1 genes. This score classifies a patient as being at "low", "intermediate-1", "intermediate-2" or "high" risk. The four aforementioned mutations are considered "high risk" according to the ELN 2018. The ICC 2022 also assigns a pejorative prognostic value to NPM1 mutations in CMML with a high risk of acutisation. NPM1 and FLT3 mutations are therefore also analysed, because even though they are reported in less than 5% of CMMLs, the presence of mutations in these two genes should also lead to the diagnosis of M4-M5 AML being reconsidered. - This NGS panel also provides diagnostic support for the cytological diagnosis of CMML in blood and bone marrow. The association of TET2 and SRSF2 mutations is highly suggestive of CMML. New 2022 data from the World Health Organization (WHO) on the redefinition of the diagnostic threshold for monocyte count (¿ 0.5.109/L) will reinforce the diagnostic aid of NGS in the management of CMML, in the same way as immunophenotyping of blood monocytes. The NGS panel can also help to differentiate between the myeloproliferative (MP-CMML) and myelodysplastic (MD-CMML) forms of CMML. Mutations in the RAS (NRAS, KRAS and CBL), JAK2 and SETBP1 pathways point towards the myeloproliferative form, which has a poor prognosis. - The theranostic impact is also addressed with this panel (potential therapeutic targets: IDH1, IDH2 and FLT3).

Preanalytics

• 5 ml EDTA whole blood or 400 µL (minimum) EDTA bone marrow or DNA extracted: 1 ¿g (in a volume of 50 ¿l)
•   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
Please attach :
- the results of the last CBC/platelet count
- the bone marrow report
- the immunophenotyping results (if performed for suspected CMML - dedicated CMML Ac panel)

Specific equipment available

• S9L: Envelopes yellow for karyotypes LYON

Documents to download

• Specific request form B8-INTGB
• Panel form (DS71-INTGB-CMML)
• Informations destinées aux professionnels de santé (DS2-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)