Qualitative PCR Detection of Key Mutations in Myeloproliferative Neoplasms (MPNs)

The TRUPCR® MPN PCR Panel Kit is a real-time PCR-based in vitro diagnostic assay designed for the qualitative detection of key genetic alterations associated with myeloproliferative neoplasms (MPNs), including the JAK2-V617F, CALR (Type 1 & 2), MPL (W515K/L/A and S505N), and BCR-ABL1 (p210/p190) fusion transcripts. This multi-analyte panel combines both RNA- and DNA-based mutation analysis, utilizing specific primers and hydrolysis probes to deliver accurate and sensitive detection from peripheral blood or bone marrow samples.

The assay requires extraction of both RNA and DNA, with provided reagents for cDNA synthesis to detect RNA-based translocations (BCR-ABL1). The kit streamlines diagnostic workflows for MPN screening by combining the most clinically relevant biomarkers in a single format.

Molecular Background

Myeloproliferative neoplasms (MPNs) are clonal hematopoietic disorders characterized by overproduction of one or more blood cell lines. They are divided into:

• Philadelphia-positive (CML) — defined by BCR-ABL1 fusion
• Philadelphia-negative MPNs — including Polycythemia Vera, Essential Thrombocythemia, and Primary Myelofibrosis, often associated with mutations in JAK2, CALR, and MPL genes.
• JAK2 V617F is a somatic mutation seen in ~95% of PV and ~50–60% of ET and PMF.
• CALR mutations, especially in exon 9, are highly recurrent in JAK2-negative MPNs.
• MPL W515 mutations are common in ET and PMF and promote megakaryocyte proliferation.
• BCR-ABL1, most often associated with CML, is also present in rare AML or ALL subtypes.

These mutations contribute to pathogenesis through constitutive activation of intracellular signaling, particularly JAK-STAT pathways.

Assay Performance & Utility

The TRUPCR® MPN PCR Panel Kit enables simultaneous analysis of key diagnostic and prognostic mutations with high sensitivity:

• Detects BCR-ABL1 at 10 copies sensitivity.
• Detects JAK2, CALR, MPL mutations down to 1% mutant allele frequency.
• Provides qualitative results within a streamlined workflow.

Ideal for:

• Comprehensive MPN mutation profiling
• Differentiating Philadelphia-positive vs. -negative MPNs
• Confirming clonal markers per WHO criteria

Technical Specifications

• Assay Type: Real-time PCR (mutation detection), RT-PCR (fusion detection)
• Target Mutations:
  • JAK2 V617F (DNA)
  • CALR Type-1 (L367fs46), Type-2 (K385fs47) (DNA)
  • MPL W515K, W515L, W515A, S505N (DNA)
  • BCR-ABL1 p210 (e13a2/e14a2), p190 (e1a2) (RNA/cDNA)
• Sample Types: Peripheral blood or bone marrow
• Input Requirement: Genomic DNA and total RNA (cDNA synthesis reagents included)
• Detection Sensitivity:
  • Up to 10 copies for BCR-ABL1
  • 1% mutant allele frequency for others
• Compatible Instruments: ABI 7500, QuantStudio 3/5, Rotor-Gene Q, Bio-Rad CFX96/384, AriaMx