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Master Thesis

Détermination de la fraction mutée de NPM1 par PCR digitale dans les leucémies aiguës myéloïdes

Abstract : The mutation in the NPM1 (Nucleophosmin) gene represents one of the most frequently acquired mutations in acute myeloid leukemias (AML). In particular among AMLs with normal karyotype. Among the many types of NPM1 mutants, the type A mutation represents 80% of the mutants. The expression of mutants NPM1 constitutes a reliable molecular marker signifying the presence of residual leukemia cells. Residual disease monitoring is currently based on quantification of real-time quantitative PCR (RT-qPCR) mutated NPM1 transcripts. The results are expressed as "mutated / normal" ratio of mutants NPM1, and require the use of a reference gene. Digital PCR is a recent technology that allows the absolute quantification of a rare genetic event. It is based on the principle of micro-compartmentalisation of the sample, involving a method of dilution of the sample, followed by a conventional end-point PCR. Such an approach consists of the creation of thousands of microdroplets (ddPCR). The advantages of digital PCR are numerous: the reference gene is identical to the desired target, the sensitivity is almost identical to RQ-PCR and no use of plasmid range is required (essential in RQ-PCR). In this work, we have developed a digital droplet PCR (ddPCR), showing its feasibility on both genomic DNA and complementary DNA. At the same time, we were able to validate the applicability of the probe designed for the detection of NPM1 mutation variant A. ddPCR has proven to be a feasible and attractive method for longitudinal monitoring of mutated NPM1 AML patients.
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Master Thesis
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Submitted on : Tuesday, October 16, 2018 - 10:40:44 AM
Last modification on : Wednesday, July 15, 2020 - 8:58:04 AM
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  • HAL Id : dumas-01896439, version 1


Arbab Ahmadreza. Détermination de la fraction mutée de NPM1 par PCR digitale dans les leucémies aiguës myéloïdes. Médecine humaine et pathologie. 2018. ⟨dumas-01896439⟩



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