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 electrophoresis. FLT3 mutation status is determined by the LeukoStrat The LeukoStrat CDx FLT3 Mutation Assay is a PCR-based in vitroCDx FLT3 Software. A FLT3 ITD and/or TKD mutation is reported as diagnostic test designed to detect internal tandem duplicationsPositive if the mutant:wild-type signal ratio meets or exceeds the clinical (ITD) and tyrosine kinase domain (TKD) mutations D835 and I836cutoff of 0.05. 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 (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 withprimers that are in and around the JM region. Wild-type FLT3 alleles AML for whom RYDAPT (midostaurin) treatment is being considered. will amplify and produce a product at 3271 bp as measured by this assay, while alleles that contain ITD mutations will produce a product In regions where gilteritinib fumarate is available, the LeukoStrat CDxthat exceeds 3271 bp (see Figure, right). FLT3 Mutation Assay is used as an aid in the assessment of patients 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 PCR and then an EcoRV restriction digest is performed. Wild-type Summary and Explanation of the Test alleles of the FLT3 gene yield digestion products of 791 bp whereas mutant alleles yield products of 1251 bp or 1271 bp from the original AML in general has a poor prognosis. Assessment of the mutation statusundigested amplicon product of 1451 bp or 1471 bp, as measured by of the FLT3 (fms related tyrosine kinase 3) receptor gene in karyotypethis assay (please see Figure, right).normal AML is the most important prognostic indicator of disease outcome, which is often substantial, as many studies in AML have shown that the presence of FLT3 activating mutations portends a poorReferencesprognosis. 1,2The LeukoStrat CDx FLT3 Mutation Assay targets regions of1. Murphy KM et al., A Clinical PCR/Capillary Electrophoresis Assay for the FLT3 gene to identify ITD mutations and TKD mutations, such as thethe Detection of Internal Tandem Duplication and Point Mutation of D835 and I836 mutations, and has been validated in an internationalthe FLT3 Gene. J. Mol. Diag. 5:96-102 (2003).clinical trial. 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. DNA is amplified via PCR and the amplicons are detected via capillary 90'