b'Gel & Capillary | IVD Assays CDx FLT3 Mutation Assay (CE-marked)Gel and Capillary Assays andAssaysCE-marked AssayCDx FLT3 Mutation AssayThe only internationally standardized CE-IVD assay for FLT3 Signal Ratio mutation analysis for assessment of acute myeloid leukemia (AML) patients eligible for treatment with RYDAPT (midostaurin) or XOSPATA (gilteritinib fumarate).Intended Use DNA is amplified via PCR and the amplicons are detected via capillary The LeukoStrat CDx FLT3 Mutation Assay is a PCR-based in vitroelectrophoresis. FLT3 mutation status is determined by the LeukoStrat diagnostic test designed to detect internal tandem duplicationsCDx FLT3 Software. A FLT3 ITD and/or TKD mutation is reported as (ITD) and tyrosine kinase domain (TKD) mutations D835 and I836Positive if the mutant:wild-type signal ratio meets or exceeds the clinical in the FLT3 gene in genomic DNA extracted from mononuclear cellscutoff of 0.05. obtained from peripheral blood or bone marrow aspirates of patientsMethod Descriptiondiagnosed with acute myelogenous leukemia (AML).ITD Mutations of FLT3 In regions where midostaurin is available, the LeukoStrat CDx FLT3The LeukoStrat CDx FLT3 Mutation Assay uses fluorescently labeled Mutation Assay is used as an aid in the assessment of patients with AML for whom RYDAPT (midostaurin) treatment is being considered. primers that are in the JM region. Wild-type FLT3 alleles will amplify and produce a product at 3271 bp as measured by this assay, while In regions where gilteritinib fumarate is available, the LeukoStrat CDxalleles that contain ITD mutations will produce a product that exceeds FLT3 Mutation Assay is used as an aid in the assessment of patients3301 bp (please see Figure, right).with AML for whom XOSPATA (gilteritinib fumarate) treatment isTKD Mutations of FLT3being considered.The LeukoStrat CDx FLT3 Mutation Assay uses primers that lie on either side of the TKD region. The FLT3 target region is amplified using Summary and Explanation of the Test PCR and then an EcoRV restriction digest is performed. Wild-type alleles of the FLT3 gene yield digestion products of 791 bp whereas AML in general has a poor prognosis. Assessment of the mutation statusmutant alleles yield products of 1251 bp or 1271 bp from the original of the FLT3 (fms related tyrosine kinase 3) receptor gene in karyotypeundigested amplicon product of 1451 bp or 1471 bp, as measured by normal AML is the most important prognostic indicator of diseasethis assay (please see Figure, right).outcome, which is often substantial, as many studies in AML have shown that the presence of FLT3 activating mutations portends a poor 1,2 Referenceprognosis.The LeukoStrat CDx FLT3 Mutation Assay targets regions of the FLT3 gene to identify ITD mutations and TKD mutations, such as the1. Murphy KM et al., A Clinical PCR/Capillary Electrophoresis Assay for D835 and I836 mutations, and has been validated in an internationalthe Detection of Internal Tandem Duplication and Point Mutation of clinical trial. the FLT3 Gene. J. Mol. Diag. 5:96-102 (2003).2. Yamamoto, Y., et al., Activating mutation of D835 within the The LeukoStrat CDx FLT3 Mutation Assay includes reagents, equipment,activation loop of FLT3 in human hematologic malignancies. Blood, software and procedures for isolating mononuclear cells and extracting97(8):2434-9 (2001).DNA from patient specimens to determine if FLT3 mutations are present. 88'