Hematological Malignancies
Thrombocytosis Panel
Background:
The majority of patients with BCR-ABL-negative myeloproliferative neoplasms (MPNs) include polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). These patients carry an acquired mutation in either JAK2, CALR or MPL and these mutations are usually mutually exclusive. Screening for these mutations not only helps to establish a diagnosis but also have an impact on the clinical outcome of these neoplasms.
Reasons for Referral:
The JAK2, CALR and MPL testing is done to molecularly subclassify myeloproliferative neoplasms, which is important in making the correct diagnosis as well as determining prognosis, and, in some cases, 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, hotspot exon 9 in CALR, and hotspot exon 10 in MPL 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; CALR and MPL are 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:
7-14 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: