b'Gel & Capillary | IVD Assays CDx FLT3 Mutation Assay - AUSGel and Capillary Assays andAssaysARTG-Approved Assay Available in AustraliaCDx FLT3 Mutation AssayThe only internationally standardized IVD assay for FLT3 Signal Ratio mutation analysis for selection of acute myeloid leukemia (AML) patients eligible for treatment with midostaurin.Intended Use Method DescriptionThe LeukoStrat CDx FLT3 Mutation Assay is a PCR-based in vitroITD Mutations of FLT3 diagnostic test designed to detect internal tandem duplicationsThe LeukoStrat CDx FLT3 Mutation Assay uses fluorescently labeled (ITD) and tyrosine kinase domain (TKD) mutations D835 and I836primers that are in the JM region. Wild-type FLT3 alleles will amplify in the FLT3 gene in genomic DNA extracted from mononuclear cellsand produce a product at 3271 bp as measured by this assay, while obtained from peripheral blood or bone marrow aspirates of patientsalleles that contain ITD mutations will produce a product that exceeds diagnosed with acute myelogenous leukemia (AML).3301 bp (please see Figure, right). In regions where midostaurin is available, theLeukoStrat CDx FLT3TKD Mutations of FLT3Mutation Assay is used as an aid in the selection of patients with AML for whom midostaurin treatment is being 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 haveReferenceshown that the presence of FLT3 activating mutations portends a poor prognosis. 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 activation loop of FLT3 in human hematologic malignancies. Blood, The LeukoStrat CDx FLT3 Mutation Assay includes reagents, equipment,97(8):2434-9 (2001).software and procedures for isolating mononuclear cells and extracting DNA from patient specimens to determine if FLT3 mutations are present. DNA is amplified via PCR and the amplicons are detected via capillary electrophoresis. FLT3 mutation status is determined by the LeukoStrat CDx FLT3 Software. A FLT3 ITD and/or TKD mutation is reported as Positive if the mutant:wild-type signal ratio meets or exceeds the clinical cutoff of 0.05. 90'