b'Gel & Capillary | CDx IVD Assay - JapanCDX FLT3Gel and Capillary Assays andAssaysCDX 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 onDepicted is a representation of the FLT3 juxtamembrane (JM) region (TM = transmembrane) and the activating loop of the tyrosine kinase Summary and Explanation of the Test either side of the TKD region. The FLT3 target region is amplified(TK) domain. Black arrows represent the relative positions of primers that target in and around the JM region for ITD or the activating loop of using PCR and then an EcoRV restriction digest is performed. Wild- the kinase domain for TKD. Colored dots represent fluorophores on labeled primers. The yellow box has vertical black lines that represent the AML in general has a poor prognosis. Assessment of the mutationtype alleles of the FLT3 gene yield digestion products of 791 bpposition of the EcoRV restriction digest sites.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). Reagentshave shown that the presence of FLT3 activating mutations portends a poor prognosis. 1,2The LeukoStrat CDx FLT3 Mutation Assay targetsReagent Name Units in Assayregions of the FLT3 gene to identify ITD mutations and TKD mutations,References FLT3 Extraction Control 1 x 1800 L tubesuch as the D835 and I836 mutations, and has been validated in an1. Murphy KM et al., A Clinical PCR/Capillary Electrophoresis AssayFLT3 ITD Master Mix 1 x 1500 L tubeinternational clinical trial. for the Detection of Internal Tandem Duplication and Point Mutation of the FLT3 Gene. J. Mol. Diag. 5:96-102 (2003). FLT3 TKD Master Mix 1 x 1500 L tubeThe LeukoStrat CDx FLT3 Mutation Assay includes reagents,2. Yamamoto, Y., et al., Activating mutation of D835 within the FLT3 ITD Positive Control 1 x 100 L tubeequipment, software and procedures for isolating mononuclear cellsactivation loop of FLT3 in human hematologic malignancies. and extracting DNA from patient specimens to determine if FLT3Blood, 97(8):2434-9 (2001). FLT3 TKD Positive Control 1 x 100 L tubemutations are present. DNA is amplified via PCR with fluorescentlyFLT3 No Template Control 1 x 200 L tubeTaq DNA Polymerase Enzyme 1 x 200 L tubeEcoRV Enzyme 1 x 200 L tubeAll reagents should be stored at -15 to -30 degrees C.Ordering InformationCatalog # Products QuantityK-412-0331 LeukoStrat CDx FLT3 Mutation Assay (Japan) 33 reactionsK-412-0341 LeukoStrat CDx FLT3 Mutation Assay Software (Japan) 1 CD complimentary with purchaseThese are in vitro diagnostic products, and are available for sale or use within Japan.92Invivoscribe 2021|93'