Background:
The majority of patients with chronic myeloproliferative neoplasms (MPNs)- polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF) carry an acquired V617F point mutation in the JAK2 kinase and in some patients, a lower frequency of JAK2 exon 12 mutation. Screening for these mutations in patients with myeloproliferative lesions in the bone marrow not only helps to establish a diagnosis but is also predicted to impact the clinical outcome of these neoplasms.
Reasons for Referral:
The JAK2 mutation testing is done on BCR-ABL negative myeloproliferative neoplasms for molecularly sub-classification and is consistent with the diagnosis of clonal hematopoeitic myeloproliferative stem cell disorder. It is also important in predicting responses to targeted and/or non-targeted therapy.
Methodology:
This test is performed by PCR-based Next Generation Sequencing of DNA extracted from peripheral blood or bone marrow. The JAK2 V617F exon and hotspot JAK2 exon 12 are sequenced using massively parallel sequencing (next-generation sequencing) with a combination of multiplexed PCR (customized QIAseq Targeted DNA panel) and sequencing on an Illumina platform. An in-house bioinformatics analysis pipeline has been used that employs multiple established variant calling tools (FreeBayes, MuTect2 and Scalpel) and variant annotation tools. The genomic variants have been interpreted in accordance with the 2017 guideline recommendations by AMP/ASCO/CAP (PMID: 27993330). The assay is validated in accordance with the AMP guidelines (PMID: 28341590).
Sensitivity:
The lower detection limit for JAK2 V617F is 1% VAF and JAK2 exon 12 is 1-2% VAF depending on read quality and quantity.
Specimen Requirements:
- Blood: 5-7mL EDTA or ACD (Solution A or B)
- Bone Marrow: 2-3mL EDTA or ACD (Solution A or B)
- DNA: 200ng at a minimum of 25ng/µL (DNA must be extracted in a CLIA-certified laboratory or a laboratory meeting equivalent requirements as determined by the CAP and/or CMS)
A REQUISITION FORM MUST ACCOMPANY ALL SAMPLES. Please include detailed clinical information.
Test Performed (Days):
Weekly
Turn Around Time:
7-10 days
Shipment Sensitivity Requirements:
Package and ship specimen to remain cold, but not frozen.
Ship via overnight express, using the FedEx priority overnight label provided.
Contact Client Services for shipping kits and instructions at (855) 535-1522.
References:
Additional Info: