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 an aid in the selection of patients with AML for whom RYDAPT® (midostaurin) treatmentis being considered. • As a tool to identify AML patients eligible for treatment with XOSPATA® (gilteritinib fumarate). Interpretation Turn-around Time Specimen Requirements Shipping Conditions Storage Conditions An interpretive report will be issued, indicating whether the patient is eligible for midostaurin or gilteritinib 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 | 13 LeukoStrat ® CDx FLT3 Mutation Assay - USA 12 Predictive test for the efficacy of midostaurin and gilteritinib fumarate Intended Use The LeukoStrat CDx FLT3 Mutation Assay is a PCR-based in vitro diagnostic test designed to detect internal tandem duplications (ITD) and tyrosine kinase domain (TKD) mutations D835 and I836 in the FLT3 gene in genomic DNA extracted from mononuclear cells obtained from peripheral blood or bone marrow aspirates of patients diagnosed with acute myelogenous leukemia (AML). The LeukoStrat CDx FLT3 Mutation Assay is used as an aid in the selection of patients with AML for whom RYDAPT® (midostaurin) treatment is being considered. The LeukoStrat CDx FLT3 Mutation Assay is used as an aid in the selection of patients with AML for whom XOSPATA® (gilteritinib) treatment is being considered. The LeukoStrat CDx FLT3 Mutation Assay is to be performed only at Laboratory for Personalized Molecular Medicine (LabPMM) LLC, a single laboratory site, located at 10222 Barnes Canyon Rd., Bldg. 1, San Diego, CA 92121. 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 Midostaurin was approved by the FDA for the treatment of adult patients with newly diagnosed AML who are FLT3 mutation positive The LeukoStrat® CDx FLT3 Mutation Assay is the first FDA approved (PMA# P160040) predictive test for the efficacy of midostaurin therapy in all AML patients, regardless of cytogenetics Gilteritinib fumarate was approved by the FDA to identify AML patients eligible for treatment with (XOSPATA® ) gilteritinib fumarate.