Overall, our results demonstrate that the use of the LAIP-based DfN-method improves the accuracy of MFC-MRD analysis in AML patients with the NPM1 mutation, and emphasize the key role of differentiated evaluation of MRD samples according to therapeutic setting and the definition of different cut-offs to improve MFC-MRD performance. The gene discussed is NPM1; the disease is acute myeloid leukemia.