Intended Use The LeukoStrat CDx FLT3 Mutation Assay is a PCR- based, in vitro diagnostic test designed to detect internal tandem duplication (ITD) mutations and tyrosine kinase domain (TKD) mutations in genomic DNA extracted from mononuclear cells obtained from peripheral blood or bone marrow aspirates of patients diagnosed with acute myelogenous leukemia. The LeukoStrat CDx FLT3 Mutation Assay is used as an aid in the assessment of patients with AML for whom Gilteritinib Fumarate treatment is being considered. LeukoStrat ® CDx FLT3 Mutation Assay - Japan Mutations in the FLT3 gene are the most common mutations found in AML Presence of a FLT3 mutation in patients with AML is both highly prognostic and clinically actionable Gilteritinib fumarate received manufacturing and marketing approval for the treatment of FLT3 mutation-positive relapse of refractory AML in Japan The LeukoStrat® CDx FLT3 Mutation Assay is the first PMDA approved test for assessment of AML patients eligible for treatment with gilteritinib fumarate 16 Test Name LeukoStrat® CDx FLT3 Mutation Assay Assay Type Capillary Electrophoresis Method Description The LeukoStrat® CDx FLT3 Mutation Assay is designed to detect ITD and TKD mutations in the FLT3 gene. The assay is performed on DNA isolated from mononuclear cells obtained from peripheral blood or bone marrow aspirates of patients diagnosed with AML. Primers targeting both the juxtamembrane region for ITD testing and kinase domain of the FLT3 gene are used to amplify DNA extracted from a patient sample. The TKD PCR product is further digested with a restriction enzyme. The ITD PCR products and the digested TKD PCR products are analyzed on a capillary electrophoresis instrument. FLT3 ITDs are detected by a change in the expected size of a wild type fragment. An amplicon larger than the wild type fragment indicates the presence of FLT3 ITD. The TKD digestion pattern identifies loss of the normal gene sequences and ensures that digestion occurred. Indications for Testing • At initial diagnosis of AML •  As a tool for the assessment of AML patients for whom gilteritinib fumarate treatment is being considered Interpretation Turn-around Time Specimen Requirements Shipping Conditions Storage Conditions An interpretive report will be issued, indicating whether the patient is eligible for gilterinib fumarate treatment 2 to 3 business days •  3 mL of peripheral blood in sodium heparin tubes only • 1 mL of bone marrow in sodium heparin tubes only  2°C to 8°C up to 72 hours; Do not freeze.  2°C to 8°C up to 7 days CDx Testing LabPMM Services Catalog 2019 | 17 Predictive test for the efficacy of Gilteritinib Fumarate