Abstract
Background
Nucleophosmin 1 (NPM1) is one of the most commonly mutated genes in acute myeloid leukemia, with mutations observed in approximately 30% of all adult cases. The persistence of NPM1 mutations following chemotherapy is associated with a greater risk of relapse as well as a lower rate of survival, making NPM1 measurable residual disease (MRD) an informative clinical target.
Methods
Herein, we have developed a straightforward unique molecular identifier (UMI)-based amplicon next-generation sequencing method for the detection of NPM1-mutated MRD that addresses some of the limitations present in other assays.
Results
The NPM1 assay allowed for accurate counting of individual mutant and wild-type molecules down to 0.01% variant allelic frequency. In silico contamination experiments highlighted the ability of this UMI methodology to maximize specificity through dramatic reductions in sequencing/demultiplexing bleed-through error.
Conclusion
Performance and clinical utility of the NPM1 MRD assay are established via both validation experiments and analyses of live performance over 1.5 years of routine clinical service.
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Conflict of interest
JPS has received consulting fees for molecular diagnostics advisory boards from Bristol Myers Squibb, Novartis, Merck, AstraZeneca, and AbbVie, and has received grant funds from AbbVie for development of other NGS-based measurable residual disease assays. LLR has received honoraria for molecular diagnostics advisory boards from Bristol Myers Squibb, AbbVie, Loxo Oncology, and Personal Genome Diagnostics. MP, CJZ, MNW, RP, CMH, NZJ, MJH, RH, PW, LVF, and SK have no conflicts of interest to declare.
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No funding was received for the conduct of this study.
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Informed consent was waived as only retrospective blood and bone marrow samples obtained for diagnostic purposes were used for this study.
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Ritterhouse, L.L., Parilla, M., Zhen, C.J. et al. Clinical Validation and Implementation of a Measurable Residual Disease Assay for NPM1 in Acute Myeloid Leukemia by Error-Corrected Next-Generation Sequencing. Mol Diagn Ther 23, 791–802 (2019). https://doi.org/10.1007/s40291-019-00436-8
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DOI: https://doi.org/10.1007/s40291-019-00436-8