High-Resolution Melting Point Assay for Rapid, Simultaneous Detection of JAK2, MPL and CALR Variants
Rapid detection of gene variants in myeloproliferative neoplassms.
Introduction
Sande et al. conducted a comprehensive evaluation of the clinical utility of the HRM-based HemeScreen MPN assay as a generic diagnostic approach for Philadelphia-negative myeloproliferative neoplasms (MPNs). The majority of mutations identified in MPNs are the canonical JAK2 p.V617F, with fewer cases presenting CALR exon 9 mutations, and even fewer carrying MPL exon 10 or JAK2 exon 12 mutations. There is a growing demand for a test capable of simultaneously detecting all prevalent mutations found in MPNs. High-resolution melting (HRM) emerges as an economically efficient and highly reliable diagnostic technique, facilitating multiplex testing.
Main Points
- Paired peripheral blood (90%) and bone marrow aspirate (10%) specimens were analysed by HRM and Sanger seguencing for gene all targets.
- All positive cases detected by HRM were confirmd by Sanger sequencing.
- The study cohort showed 14% of specimens carrying 139 variants: 114 were single nucleotide variants, 3 insertions (5bp), 21 deletions (3-52bp) and 1 insertion/deletion with a VAF from <5% to 95%.
- The HRM assay overall had a sensitivity of 96.7% and specificity of 98.7% with a PPV of 92.3% and NPV of 99.5%.
Additionally, variants not known to be associated with the disease were identified by HRM and confirmed through Sanger sequencing.
Conclusions
The HRM-based MPN assay demonstrated a 99.5% NPV in a large real-world patient cohort. Its clinical performance, accuracy, cost-effectiveness, and detection of recurrent somatic variants render the HRM-based platform an outstanding screening tool.
Epistem Services
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