Acute Myeloid Leukaemia (AML)/ Myelodysplastic Syndrome (MDS)

Myelodysplastic syndrome (MDS) is a group of diseases that affects normal blood cell production in the bone marrow. About 30% of people with MDS will progress to Acute Myeloid Leukaemia (AML). AML are diseases arising from myeloid cells (mostly white blood cells other than lymphocytes) and is more common in older people[1-2].

Mutations in the FLT3 gene (FLT3-ITD or FLT3-TKD) occur in approximately 25% to 30% of AML cases. Presence of FLT3 mutations is associated with a particularly aggressive form of the disease and shorter overall survival. FDA has approved the use of targeted drug, Midostaurin for AML patients with FLT3 mutations. Therefore, testing for FLT3 mutations is important to provide optimal treatment care for AML patients[2-4].

Recurring mutations in isocitrate dehydrogenase (IDH) genes are detected in approximately 20% of adult patients with AML. Several studies have suggested that the presence of these mutations confers an adverse prognosis. FDA has recently approved to use of targeted agent enasidenib for adults with relapsed or refractory AML associated with IDH2 mutations. On the other hand, mutations in other genes including NPM1 and CEPBA indicate good prognosis[2-3].

Detection of RUNX1/RUNX1T1, CBFB/MYH11 and PML/RARA fusion helps in diagnosis of AML. The presence of these fusions indicates good prognosis. However, the presence of KIT mutations (in exon 8 & 17) among patients with RUNX1/RUNX1T1 and CBFB/MYH11 fusions will have adverse impact on prognosis of the patients. Therefore, results of both fusion and mutation should be known for proper treatment. In addition, the presence of BCR/ABL1 fusions indicates adverse prognosis but also represents the availability of Tyrosine Kinase Inhibitor (TKI) treatment[2-3].

Identification of gene mutations or fusions in AML is important in diagnosis and therapeutic decision-making in AML as different mutations and fusions in different genes have different influences on treatment and prognosis. Oncode AML panel offers range of PCR and NGS panels to detect mutations and fusions in AML.

AML/ MDS Mutation (DNA Assays)

ONCODEduce AML PCR Panel
ONCODEduce AML PCR Panel are PCR based qualitative assays with amplicon identification by fragment analysis to detect mutation in FLT3-ITD & FLT3-TKD D835/I836 and NPM1. The FLT3-TKD assay employs restriction enzyme digestion to detect the D835 & I836 mutations. The detection limit of FLT3-TKD & NPM1 assays is ~5%; however, it is difficult to estimate detection limit for the FLT3-ITD assay given the variable size of the duplication.

Specimen Requirements:
Blood: 3ml in EDTA OR Marrow Aspirate: 1-2.5ml in EDTA

Turn Around Time: • 10 working days

Shipment Requirement:
• Keep specimen at room temperature. DO NOT expose to direct sunlight.
ONCODEduce AML Core NGS Panel
ONCODEduce AML Core (NGS) Panel includes a core set of 11 genes implicated in AML and with targeted therapy namely FLT3-ITD & TKD, NPM1, CEBPA, IDH1, IDH2, TP53, ASXL1, DNMT3A, KIT, JAK2 & RUNX1.

Specimen Requirements:
Blood: 3ml in EDTA OR Marrow Aspirate: 1-2.5ml in EDTA OR Aspirate: 2 smears with adequate cellularity

Turn Around Time:
• 10 working days

Shipment Requirement:
• Keep specimen at room temperature. DO NOT expose to direct sunlight.
ONCODEduce AML Comprehensive NGS Panel
ONCODEduce AML Comprehensive NGS Panel allow a broader exploration of the genes implicated in AML/MDS by targeting 37 genes . This assay has a detection limit of ~1%.

Specimen Requirements:
Blood: 3ml in EDTA OR Marrow Aspirate: 1-2.5ml in EDTA OR Aspirate: 2 smears with adequate cellularity

Turn Around Time:
• 15 working days

Shipment Requirement:
• Keep specimen at room temperature. DO NOT expose to direct sunlight.
ONCODEduce AML/MDS Extended NGS Panel
ONCODEduce AML/MDS Extended (NGS) Panel Panels allow a broader exploration of the genes implicated in AML/MDS by targeting 73 genes . This assay has a detection limit of ~1%.

Specimen Requirements:
Blood: 3ml in EDTA OR Marrow Aspirate: 1-2.5ml in EDTA OR Aspirate: 2 smears with adequate cellularity

Turn Around Time:
• 15 working days

Shipment Requirement:
• Keep specimen at room temperature. DO NOT expose to direct sunlight.

AML Fusion (RNA Assay)

ONCODEduce AML Fusion PCR Panel
ONCODEduce AML Fusion PCR Panel is a qualitative PCR assay to detect the commonest rearrangements in AML by targeting 10 gene fusions including RUNX1/RUNX1T1, CBFB/MYH11, PML/RARA and BCR/ABL1.
 
BCR/ABL1 KMT2A/ELL NPM1/MLF1 RUNX1/RUNX1T1
CBFB/MYH11 KMT2A/MLLT3 PICALM/MLLT10 KMT2A/PTD (MLL/PTD) *
DEK/NUP214 KMT2A/MLLT4 PML/RARA
* partial tandem duplication

Specimen Requirements:
Blood: 3ml in EDTA OR Marrow Aspirate: 1-2.5ml in EDTA

Turn Around Time:
• 15 working days

Shipment Requirement:
• Keep specimen at room temperature. DO NOT expose to direct sunlight.
ONCODEduce AML Fusion Core & KIT Mutations NGS Panel
ONCODEduce AML Fusion Core & KIT Mutations NGS Panel includes the commonest rearrangements in AML targeting 12 genes including RUNX1, CBFB, RARA, BCR, ABL1, MLL/KMT2A fusions. In addition, this assay detects mutations in exon 8 & 17 of KIT, important in the management of patients with RUNX1/RUNX1T1 & CBFB/MYH11 fusions.

Specimen Requirements:
Blood: 3ml in EDTA OR Marrow Aspirate: 1-2.5ml in EDTA OR Aspirate: 2 smears with adequate cellularity

Turn Around Time:
• 15 working days

Shipment Requirement:
• Keep specimen at room temperature. DO NOT expose to direct sunlight.
ONCODEduce AML Fusion Comprehensive NGS Panel
ONCODEduce AML Fusion Comprehensive NGS Panel is designed to detect almost all fusions implicated in AML by targeting 87 genes .

Specimen Requirements:
Blood: 3ml in EDTA OR Marrow Aspirate: 1-2.5ml in EDTA OR Aspirate: 2 smears with adequate cellularity

Turn Around Time:
• 15 working days

Shipment Requirement:
• Keep specimen at room temperature. DO NOT expose to direct sunlight.
References;
1. Leukaemia Foundation (2017). Myelodysplastic syndrome (MDS). READ MORE
2. Seegmiller, A., Jagasia, M., Wheeler, S. Vnencak-Jones, C. (2016). Molecular Profiling of Acute Myeloid Leukemia. My Cancer Genome. READ MORE
3. Taylor, J., Xiao, W., & Abdel-Wahab, O. (2017). Diagnosis and Classification of Hematologic Malignancies on the Basis of Genetics. Blood, blood-2017.
4. Levis, M. (2013). FLT3 Mutations in Acute Myeloid Leukemia: What Is The Best Approach in 2013?. ASH Education Program Book 1, 220-226.