Test Name NPM1 mutation analysis (qualitative) Assay Type Capillary Electrophoresis Method Description Primers targeting exon 12 on the NPM1 gene are used to amplify the patient’s DNA. The size of the NPM1 PCR product is determined by capillary electrophoresis. LabPMM offers the only internationally harmonized assay for NPM1 mutations and testing is performed pursuant to patents licensed from TrovaGene, Inc. of San Diego, CA. Indications for Testing • At initial diagnosis of AML • Stratification high and low risk AML •  Recurrence of leukemia after induction therapy on patients not initially screened for NPM1 mutations Interpretation Turn-around Time Specimen Requirements Shipping Conditions Storage Conditions An interpretive report will be issued indicating whether a NPM1 mutation was detected 1-3 business days •  3 mL peripheral blood in EDTA, ACD or Heparin •  1 mL bone marrow in EDTA, ACD or Heparin •  250 ng of previously isolated DNA  Ambient or Cool;  Do not freeze • Room Temp up to 72 hours •  2-8 °C up to 7 days Molecular Diagnostic Tests LabPMM Services Catalog 2019 | 27 Clinical Information The Nucleophosmin (NPM1) gene is one of the most commonly mutated genes in acute myeloid leukemia (AML), occurring in about 35% of AML patients at diagnosis1 . The vast majority of NPM1 mutations are insertions in exon 12 occurring near the C-terminus of the protein that result in cytoplasmic localization2 . Currently there are over 40 known NPM1 mutations, most of which will be detected with our assay. Clinical studies have found that NPM1 mutations are associated with increased blast counts, higher extramedullary involvement and increased platelet counts in AML3 . Furthermore, in the absence of a FLT3 ITD mutation (or FLT3 ITD with a low ratio), NPM1 mutations are associated with a favorable prognosis4 . It has been suggested that the identification of mutations in both NPM1 and FLT3 genes allows for the stratification of the AML patients into three different prognostic groups: • Favorable prognosis: NPM1 mutation without FLT3 ITD or with FLT3 ITDlow • Intermediate prognosis: NPM1 mutation and FLT3 ITDhigh ; NPM1- without FLT3 ITD or with FLT3 ITDlow (without adverse-risk genetic lesions) • Poor prognosis: NPM1 wild-type and FLT3 ITDhigh It is recommended that AML patients be screened for NPM1 mutations as an effort to assess prognosis and aid in treatment decisions. Results from NPM1 and FLT3 mutational screening should be available within 48 to 72 hours (at least in patients eligible for intensive chemotherapy). Utilizing both NPM1 and FLT3 (mutant:wild-type ratio) mutation status is the most common molecular method for stratification of the AML population. NPM1 References 1.  Thiede C, et al. (2006) Prevalence and prognostic impact of NPM1 mutations in 1485 adult patients with acute myeloid leukemia (AML). Blood 107:4011-4020. 2.  Falini B. et al. (2007) Translocations and mutations involving the nucleophosmin (NPM1) gene in lymphomas and leukemias. Haematologica 92(4):519-532. 3.  Döhner K, et al. (2005) Mutant nucleophosmin (NPM1) predicts favorable prognosis in younger adults with acute myeloid leukemia and normal cytogenetics: interaction with other gene mutations. Blood 106(12):3740-3746. 4.  Döhner H, et al. (2017) Diagnosis and management of AML in adults: 2017 ELN recommendations from an international expert panel. Blood 129:424-447. 26