Quantitative PCR Detection of JAK2 V617F Allele Burden in Myeloproliferative Neoplasms

The TRUPCR® JAK2 V617F Allele Burden Kit is a CE-IVD certified, real-time PCR assay developed for the quantification of the V617F mutation in the JAK2 gene from genomic DNA. Designed to measure the mutant allele burden in patients with myeloproliferative neoplasms (MPNs), this assay enables precise quantification of JAK2 V617F copies against a wild-type DNA background. The kit utilizes mutation-specific primers and a JAK2 V617F-targeted hydrolysis probe, achieving a detection limit as low as 1% mutant DNA. It is calibrated against the WHO 1st International Reference Panel and is compatible with standard real-time PCR platforms.

Molecular Background

JAK2 (Janus Kinase 2) is a non-receptor tyrosine kinase that plays a central role in cytokine signaling and hematopoietic cell regulation. It activates intracellular pathways via phosphorylation of downstream targets like STAT5, promoting cell proliferation, differentiation, and survival.

A single nucleotide mutation in JAK2—c.G1849T; p.Val617Phe (V617F)—is one of the most common somatic mutations associated with BCR-ABL1-negative MPNs. This gain-of-function mutation results in constitutive activation of JAK-STAT signaling, driving clonal expansion of hematopoietic cells.

The JAK2 V617F mutation is found in:

• 95–98% of polycythemia vera (PV) cases
• 50–60% of essential thrombocythemia (ET) and primary myelofibrosis (PMF)
• Occasionally in other myeloid neoplasms, such as chronic myelomonocytic leukemia and myelodysplastic syndrome

Detection of this mutation is included in the WHO diagnostic criteria for MPNs and is essential for diagnosis, risk stratification, and treatment monitoring. JAK2 allele burden may also decrease with successful therapy and reappear during relapse, making it a valuable tool for disease tracking.

Assay Performance & Utility

The TRUPCR® JAK2 V617F Allele Burden Kit delivers accurate, reproducible quantification of JAK2 mutant DNA in a background of wild-type DNA. The assay is optimized to:

• Detect as low as 1% mutant allele
• Quantify allele burden for treatment monitoring and risk assessment
• Provide high specificity using site-directed hydrolysis probes
• Align with WHO standards for clinical consistency

This assay is ideal for use in clinical molecular laboratories, hematology/oncology centers, and monitoring post-transplant or therapy response in MPN patients.

Technical Specifications

• Assay Type: Real-time PCR (quantitative)
• Target Mutation: JAK2 V617F (c.G1849T; p.Val617Phe)
• Sample Type: Genomic DNA from whole blood
• Input Requirement: 50–200 ng per reaction
• Sensitivity: Detects 1% mutant allele fraction
• Probe Chemistry: Mutation-specific hydrolysis probes
• Compatible Instruments: ABI 7500, Roche LightCycler®, Bio-Rad CFX96™, and others