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Lab tests for MPN
Alice Moncada1, Alessandro Pancrazzi1
1Laboratory Medicine Department, Molecular and Clinical Pathology Sector, Azienda USL Toscana Sudest, Ospedale San Donato, Arezzo, Italy.
International Review of Cell and Molecular Biology
|February 14, 2022
Summary
Molecular investigations for myeloproliferative neoplasms (MPN) detect BCR-ABL for chronic myeloid leukemia and driver gene variants for Philadelphia-negative MPN. Next-generation sequencing aids in diagnosing complex cases and determining prognosis.
Area of Science:
- Hematology
- Molecular Diagnostics
- Oncology
Background:
- Myeloproliferative neoplasms (MPN) require molecular investigations for accurate diagnosis and subtyping.
- Distinguishing between BCR-ABL positive and Philadelphia-negative MPN is critical for treatment strategies.
- Identifying specific driver gene mutations is essential for prognostication in MPN.
Purpose of the Study:
- To outline molecular laboratory investigations for MPN, covering both BCR-ABL positive and negative forms.
- To detail methods for detecting key driver gene mutations (JAK2, CALR, MPL, cKIT, FIP1L1-PDGFRalpha) in MPN.
- To highlight the role of Next-Generation Sequencing (NGS) in identifying high-risk mutations and improving prognostic accuracy.
Main Methods:
- BCR-ABL detection using RT-Polymerase Chain Reaction (RT-PCR) and droplet digital PCR (ddPCR), complemented by CBA and FISH.
- Driver gene variant analysis for MPN Ph-negative forms, including JAK2V617F, JAK2 exon 12, CALR exon 9, MPL exon 10, cKIT D816V, and FIP1L1-PDGFRalpha.
- Application of Next-Generation Sequencing (NGS) panels for comprehensive analysis of high molecular risk genes (ASXL1, EZH2, TET2, IDH1/IDH2, SRSF2) in triple-negative cases.
Main Results:
- Established methods for BCR-ABL quantification and driver gene mutation detection in various MPN subtypes.
- Demonstrated the importance of allelic burden and variant type (e.g., CALR) for prognostic assessment.
- Highlighted NGS's capability to detect subclonal mutations and high-risk variants, crucial for prognostic scoring and excluding false negatives.
Conclusions:
- Accurate molecular diagnostics are fundamental for classifying MPN and guiding therapeutic decisions.
- Quantitative analysis of driver mutations provides significant prognostic information.
- NGS represents a significant advancement for investigating complex MPN cases, especially triple-negative scenarios, and refining prognostic evaluation.