Qualitative Real-Time PCR Detection of Pediatric Leukemia Fusion Markers

The TRUPCR® Paediatric Leukemia Panel Kit is a CE-IVD certified, real-time RT-PCR assay developed for the qualitative detection of 18 diagnostic fusion gene markers and 38 transcript variants commonly associated with pediatric leukemia, including both acute lymphoblastic leukemia (ALL) and subsets of acute myeloid leukemia (AML). This multi-target panel enables precise molecular characterization of leukemic subtypes using cDNA generated from extracted RNA.

The assay supports accurate fusion transcript detection for critical gene rearrangements such as TEL-AML1, BCR-ABL1, MLL fusions, E2A-PBX1, and PAX5-JAK2, aiding in diagnostic confirmation, risk stratification, and therapeutic decision-making. The inclusion of the ABL1 gene as an endogenous internal control ensures sample quality validation and test integrity.

Molecular Background

Pediatric leukemia is the most common type of cancer in children, accounting for approximately 30% of all childhood malignancies. It primarily manifests as acute lymphoblastic leukemia (ALL) or less frequently as acute myeloid leukemia (AML). These are clonal hematologic disorders characterized by the accumulation of immature lymphoid or myeloid cells due to disruptions in normal hematopoiesis.

At the molecular level, pediatric leukemias are driven by a variety of chromosomal rearrangements and gene fusions that alter transcriptional regulation, signal transduction, or differentiation pathways. Many of these fusions serve as key diagnostic, prognostic, and therapeutic markers.

• TEL-AML1, E2A-PBX1, and TCF3-HLF are hallmark translocations in B-cell precursor ALL.
• MLL (KMT2A) rearrangements, such as MLL-AF4, MLL-AF9, and MLL-ENL, are particularly common in infant leukemias and associated with poor prognosis.
• Fusion genes involving JAK2, ABL1, or CSF1R play roles in activating tyrosine kinase signaling pathways, and are potential targets for kinase inhibitors.
• Rare but clinically relevant rearrangements like PAX5-JAK2, ETV6-ABL1, and SSBP2-CSF1R can guide risk stratification and therapy adjustment.

Accurate identification of these fusion transcripts is critical for confirming diagnosis, guiding treatment (e.g., TKI therapy in BCR-ABL1-positive cases), and determining risk groups for therapy intensification or de-escalation.

Assay Performance & Utility

• Simultaneous detection of 18 fusion gene markers and 38 known transcript variants
• Enables precise subtype classification of pediatric leukemias (ALL & select AML)
• Detects rare and cryptic rearrangements relevant to therapy and prognosis
• Minimal hands-on time with ready-to-use reagents
• Built-in quality control using ABL1 housekeeping gene
• Validated for clinical use in routine diagnostics

Technical Specifications

• Assay Type: Two-step RT-PCR (qualitative detection)
• Markers Detected: TEL-AML1, BCR-ABL1 (p190/p210/p230), E2A-PBX1, MLL fusions, P2RY8-CRLF2, TCF3-HLF, JAK2 rearrangements, and others
• Transcript Variants: 38 fusion transcript types
• Sensitivity: Detects as low as 10 copies of fusion transcript
• Sample Type: Peripheral blood or bone marrow
• Input Material: cDNA from RNA extracted samples
• Controls: Internal RNA quality control via ABL1 gene