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 labeled primers, amplicon is enzymatically digested (TKD), and the amplicons are detected via capillary electrophoresis. FLT3 mutation The LeukoStrat CDx FLT3 Mutation Assay is a PCR-based, in vitrostatus is determined by the LeukoStrat CDx FLT3 Software. A FLT3 ITD diagnostic test designed to detect internal tandem duplication (ITD)and/or TKD mutation is reported as Positive if the mutant:wild-type mutations and tyrosine kinase domain (TKD) mutations D835 and I836signal ratio meets or exceeds the clinical cutoff of 0.05.in the FLT3 gene in genomic DNA extracted from mononuclear cells obtained from peripheral blood or bone marrow aspirates of patientsMethod Descriptiondiagnosed 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 and around the JM region. Wild-type FLT3 alleles treatment is being considered. will amplify and produce a product at 3271 bp as measured by this The LeukoStrat CDx FLT3 Mutation Assay is used as an aid in theassay, while alleles that contain ITD mutations will produce a product assessment of patients with AML for whom Quizartinib Hydrochloridethat exceeds 3271 bp (see Figure, right). treatment is being considered.TKD Mutations of FLT3 The LeukoStrat CDx FLT3 Mutation Assay uses primers that lie on Summary and Explanation of the Test either side of the TKD region. The FLT3 target region is amplified using PCR and then an EcoRV restriction digest is performed. Wild-AML in general has a poor prognosis. Assessment of the mutationtype alleles of the FLT3 gene yield digestion products of 791 bp status of the FLT3 (fms related tyrosine kinase 3) receptor gene inwhereas mutant alleles yield products of 1251 bp or 1271 bp from karyotype normal AML is the most important prognostic indicator ofthe original undigested amplicon product of 1451 bp or 1471 bp, as disease outcome, which is often substantial, as many studies in AMLmeasured by this assay (see Figure, right).have shown that the presence of FLT3 activating mutations portends a poor prognosis. 1,2The LeukoStrat CDx FLT3 Mutation Assay targets regions of the FLT3 gene to identify ITD mutations and TKD mutations,Referencessuch as the D835 and I836 mutations, and has been validated in an1. Murphy KM et al., A Clinical PCR/Capillary Electrophoresis Assay international clinical trial. for the Detection of Internal Tandem Duplication and Point Mutation The LeukoStrat CDx FLT3 Mutation Assay includes reagents,of the FLT3 Gene. J. Mol. Diag. 5:96-102 (2003).equipment, software and procedures for isolating mononuclear cells2. Yamamoto, Y., et al., Activating mutation of D835 within theand extracting DNA from patient specimens to determine if FLT3activation loop of FLT3 in human hematologic malignancies. mutations are present. DNA is amplified via PCR with fluorescentlyBlood, 97(8):2434-9 (2001).94'