b'Gel & Capillary | CDx IVD Assay - JapanCDX FLT3Gel and Capillary Assays andAssaysPMDA/MHLW Approved Assay Available in JapanCDx FLT3 Mutation AssayThe only internationally standardized assay for FLT3 Signal Ratio mutation analysis for assessment of acute myeloid leukemia (AML) patients eligible for treatment with Gilteritinib Fumarate or Quizartinib Hydrochloride.Intended Use are detected via capillary electrophoresis. FLT3 mutation status is determined by the LeukoStrat CDx FLT3 Software. A FLT3 ITD and/The LeukoStrat CDx FLT3 Mutation Assay is a PCR-based, in vitroor TKD mutation is reported as Positive if the mutant:wild-type signal diagnostic test designed to detect internal tandem duplication (ITD)ratio meets or exceeds the clinical cutoff of 0.05.mutations and tyrosine kinase domain (TKD) mutations D835 and I836 in the FLT3 gene in genomic DNA extracted from mononuclear cellsMethod Descriptionobtained from peripheral blood or bone marrow aspirates of patients diagnosed with acute myelogenous leukemia. ITD Mutations of FLT3 The LeukoStrat CDx FLT3 Mutation Assay is used as an aid in theThe LeukoStrat CDx FLT3 Mutation Assay uses fluorescently labeled assessment of patients with AML for whom Gilteritinib Fumarateprimers that are in the JM region. Wild-type FLT3 alleles will amplify treatment is being considered. and produce a product at 3271 bp as measured by this assay, while alleles that contain ITD mutations will produce a product that exceeds The LeukoStrat CDx FLT3 Mutation Assay is used as an aid in the3301 bp (please see Figure, right).assessment of patients with AML for whom Quizartinib HydrochlorideTKD Mutations of FLT3treatment is being considered. The LeukoStrat CDx FLT3 Mutation Assay uses primers that lie on either side of the TKD region. The FLT3 target region is amplified Summary and Explanation of the Test using PCR and then an EcoRV restriction digest is performed. Wild-type alleles of the FLT3 gene yield digestion products of 791 bp AML in general has a poor prognosis. Assessment of the mutationwhereas mutant alleles yield products of 1251 bp or 1271 bp from status of the FLT3 (fms related tyrosine kinase 3) receptor gene inthe original undigested amplicon product of 1451 bp or 1471 bp, as karyotype normal AML is the most important prognostic indicator ofmeasured by this assay (please see Figure, right).disease outcome, which is often substantial, as many studies in AML have shown that the presence of FLT3 activating mutations portends 1,2 Referencea poor prognosis.The LeukoStrat CDx FLT3 Mutation Assay targets regions of the FLT3 gene to identify ITD mutations and TKD mutations,1. Murphy KM et al., A Clinical PCR/Capillary Electrophoresis Assay such as the D835 and I836 mutations, and has been validated in anfor the Detection of Internal Tandem Duplication and Point Mutation international clinical trial. of the FLT3 Gene. J. Mol. Diag. 5:96-102 (2003).2. Yamamoto, Y., et al., Activating mutation of D835 within theThe LeukoStrat CDx FLT3 Mutation Assay includes reagents,activation loop of FLT3 in human hematologic malignancies. equipment, software and procedures for isolating mononuclear cellsBlood, 97(8):2434-9 (2001).and extracting DNA from patient specimens to determine if FLT3 mutations are present. DNA is amplified via PCR and the amplicons 92'