Qualitative PCR Detection of JAK2 V617F Mutation in Myeloproliferative Neoplasms

The TRUPCR® JAK2 Mutation Detection Kit (V2.0) is a real-time PCR-based in vitro diagnostic assay designed for the qualitative detection of the V617F mutation in the JAK2 (Janus Kinase 2) gene from genomic DNA extracted from whole blood. The assay uses ARMS-PCR technology, enabling highly sensitive detection of the mutant allele even in the presence of a high background of wild-type DNA. It includes an internal control to verify PCR integrity and reduce false negatives, and is compatible with major real-time PCR instruments.

This kit supports the diagnosis and molecular classification of myeloproliferative neoplasms (MPNs), including polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF).

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 Mutation Detection Kit offers highly specific detection of the V617F mutation, with a limit of detection as low as 0.5% mutant allele in a background of 99.5% wild-type DNA. The assay supports:

• Early diagnosis and confirmation of MPN subtypes
• Stratification of patients for targeted therapy or disease monitoring
• Reliable result interpretation via internal reference gene control
• Compatibility with standard real-time PCR platforms

Its ARMS-PCR approach ensures allele-specific amplification, with rapid turnaround and minimal hands-on time, making it ideal for routine use in clinical molecular labs.

Technical Specifications

The kit uses allele-specific real-time PCR with Taq polymerase, which discriminates between matched and mismatched nucleotides at the 3’-end of the primer. An internal control gene is included to assess sample quality and PCR efficiency.

• Sample Type: Genomic DNA from whole blood
• Target Mutation: JAK2 V617F (c.G1849T, p.Val617Phe)
• Detection Method: ARMS-PCR via real-time PCR
• Sensitivity: Detects as low as 0.5% mutant allele
• Internal Control: Included to monitor PCR inhibition
• Compatible Instruments: ABI 7500, Roche LightCycler®, Bio-Rad CFX96™, and similar systems