b'Gel & Capillary | IVD Assays CDx FLT3 Mutation Assay (IVD)FDA Approved Companion DiagnosticCDx FLT3 Mutation Assay (IVD)FDA Approved Companion Diagnostic InVitro Diagnostic Kit|PN: K4120361InVitro Diagnostic Kit|PN: K4120361 FDA Approved Assay Available in USA Method Schematic: FLT3 ITD & TKD Mutant DetectionCDx FLT3 Mutation AssayFDA approved assay for assessment of acute myeloid leukemia (AML) patients eligible for treatment with RYDAPT (midostaurin) or XOSPATA (gilteritinib fumarate), now available as US distributed kit.This FLT3 companion diagnostic includes reagents along with software that identifies ITD and TKD mutations, generates mutant/wildtype signal ratios, and predicts response to gilteritinib and midostaurin.Clinical Significance of FLT3 Mutation Status: Each year approximately 21,000 patients in the United States are diagnosed with AML. Of those diagnosed with AML, ~1 out of 3 are expected to have presence of FLT3 mutations, (FLT3mut+). Since FLT3mut+ AML is clinically actionable, stratification of AML patients by testing for FLT3 mutation status has become a standard of care.Intended UseThe LeukoStrat CDx FLT3 Mutation Assay is a PCR-based in vitroMethod Descriptiondiagnostic test designed to detect internal tandem duplication (ITD) and tyrosine kinase domain (TKD) mutations D835 and I836ITD Mutations of FLT3 in the FLT3 gene in genomic DNA extracted from mononuclear cellsThe LeukoStrat CDx FLT3 Mutation Assay uses fluorescently labeled obtained from peripheral blood or bone marrow aspirates of patientsprimers that are in and around the JM region. Wild-type FLT3 alleles diagnosed with acute myelogenous leukemia (AML). 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 productDepicted is a representation of the FLT3 juxtamembrane (JM) region (TM = transmembrane) and the activating loop of the tyrosine kinase assessment of patients with AML for whom RYDAPT (midostaurin)that exceeds 3271 bp (see Figure, right). (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 treatment is being considered. the kinase domain for TKD. Colored dots represent fluorophores on labeled primers. The yellow box has vertical black lines that represent the TKD Mutations of FLT3 position of the EcoRV restriction digest sites.The LeukoStrat CDx FLT3 Mutation Assay is used as an aid in theThe LeukoStrat CDx FLT3 Mutation Assay uses primers that lie on assessment of patients with AML for whom XOSPATA (gilteritinib)either side of the TKD region. The FLT3 target region is amplifiedReagentstreatment is being considered.using PCR and then an EcoRV restriction digest is performed.Wild-type alleles of the FLT3 gene yield digestion products of 791 bpReagent Name Units in AssayThe test is for use on the 3500xL Dx Genetic Analyzer. whereas mutant alleles yield products of 1251 bp or 1271 bp from the original undigested amplicon product of 1451 bp or 1471 bp, FLT3 Extraction Control 1 x 1800 L tubeas measured by this assay (see Figure, right).Summary and Explanation of the Test FLT3 ITD Master Mix 1 x 1500 L tubeAcute myelogenous leukemia (AML) in general has a poor prognosis. References FLT3 TKD Master Mix 1 x 1500 L tubeMany studies in AML have shown that the presence of FLT3 activating1. Murphy KM et al., A Clinical PCR/Capillary Electrophoresis Assay forFLT3 ITD Positive Control 1 x 100 L tubemutations portends a poor prognosis making it an attractive target forFLT3 TKD Positive Control 1 x 100 L tubetreatment. 1,2 the Detection of Internal Tandem Duplication and Point Mutation of the FLT3 Gene. J. Mol. Diag. 5:96-102 (2003). FLT3 No Template Control 1 x 200 L tubeThe LeukoStrat CDx FLT3 Mutation Assay targets regions of the FLT32. Yamamoto, Y., et al., Activating mutation of D835 within theTaq DNA Polymerase 1 x 200 L tubegene to identify internal tandem duplication (ITD) mutations andactivation loop of FLT3 in human hematologic malignancies. Blood, tyrosine kinase domain (TKD) mutations, such as the D835 and I83697(8):2434-9 (2001). EcoRV Enzyme 1 x 200 L tubemutations. NEBuffer 3.1 1 x 1250 L tubeThe LeukoStrat CDx FLT3 Mutation Assay includes reagents, software and procedures for isolating mononuclear cells and extracting DNA from patient peripheral blood or bone marrow specimens to determineOrdering Informationif FLT3 mutations are present. Catalog # Products QuantityDNA is amplified via PCR and the amplicons are detected via capillary electrophoresis. FLT3 mutation status is determined by the LeukoStratK-412-0361 LeukoStrat CDx FLT3 Mutation Assay33 reactionsCDx FLT3 Software. A FLT3 ITD and/or TKD mutation is reported asK-412-0371 LeukoStrat CDx FLT3 Software CD complimentary with purchasePositive if the mutant:wild-type signal ratio meets or exceeds the clinical cutoff of 0.05. K-412-0401 LeukoStrat CDxAssay Installer USB complimentary with purchaseThese are in vitro diagnostic (IVD) products, and are available for sale or use in the United States only86Invivoscribe 2021|87'