b'Gel & Capillary | RUO AssaysGel and CapillaryFLT3 Mutation Assays Research Use Only (RUO) AssayFLT3 Mutation Assays These products are not currently available for sale or use in the United States. Assay Use Background FLT3 Mutation Assays are useful for the study of: Mutations in the fms related tyrosine kinase 3 (FLT3) gene are the most common mutations found in acute myeloid leukemia (AML), occurring Identifying FLT3 mutations in patients with AML in approximately 30% of patients at the time of diagnosis, and are Discriminating between high and low risk patients. FLT3 mutations characterized by an aggressive phenotype with a high prevalence of portend a worse prognosis for patients with AML. Therefore relapse. 1,2,3patients testing positive for FLT3 mutations may benefit from a The most prevalent and clinically significant type of FLT3 mutation is more aggressive treatment regimen an internal tandem duplication (ITD) in the juxtamembrane domain. 4Many clinical studies have found that FLT3 ITD mutations are Summary and Explanation of the Testassociated with higher concentrations of leukemic cells in both blood Acute myeloid leukemia (AML) in general has a poor prognosis. Recentand bone marrow, increased incidence of relapse, and decreased studies have described mutation of the FLT3 (fms-related tyrosineoverall survival. kinase 3) receptor to be the most important prognostic factor in AML,The second most common mutation type in the FLT3 gene is a tyrosine with FLT3 mutants having a worse outcome and response to standardkinase domain (TKD) point mutation in aspartate (D835) or isoleucine chemotherapeutic interventions. Accordingly, identification of an FLT3(I836). TKD mutations result in constitutive autophosphorylation and mutation in AML may indicate a need to reassess and modify standardactivation of FLT3. 5,6TKD mutations have been linked to poor overall treatment options. survival, but to a lesser extent as compared to ITD mutations.All types of AML can have activating mutations in the FLT3 gene. Mutation of the FLT3 receptor, either by internal tandem duplicationSpecimen Requirements (ITD) of the juxtamembrane domain or by point mutation of the aspartic acid residue (D835) or isoleucine (I836) in the activation loop of the5 cc of peripheral blood, bone marrow biopsy, or bone marrow kinase domain, causes constitutive activation of the FLT3 receptor. aspirate anti-coagulated with heparin or EDTA; or, Minimum 5 mm cube of tissue; or, This test kit includes three master mixes. The ITD and D835 master mixes2 g of genomic DNA; or, target the juxtamembrane and kinase domain regions (respectively).Formalin-fixed, paraffin-embedded tissue or slides. The third master mix, the Specimen Control Size Ladder, targets multiple genes and generates a series of amplicons of 100, 200, 300, 400, andReference600 base pairs to ensure that the quality and quantity of input DNA is adequate to yield a valid result.1. Acute Myeloid Leukemia, Clinical Practice Guidelines in Oncology, National Comprehensive Cancer Network (v.2.2014).PCR products can be analyzed by capillary electrophoresis with use2. Lowenberg, B. et al.Acute myeloid leukemia. N Engl J Med of ABI instruments or standard agarose TBE gel electrophoresis with341(14):1051-62 (1999).ethidium bromide staining. 3. Thiede, C. et al. Analysis of FLT3-activating mutations in 979 patients with acute myelogenous leukemia: association with FAB and identification of subgroups with poor prognosis. Blood 99(12): 4326-35 (2002).4. Nakao, M. et al. Internal tandem duplication of the FLT3 gene found in acute myeloid leukemia. Leukemia 10(12):1911-18 (1996).5. Yamamoto, Y et al. Activating mutation of D835 within the activation loop of FLT3 in human hematologic malignancies. Blood, 97(8):2434-9 (2001).6. Gilliland, DG et al. The roles of FLT3 in hematopoiesis and leukemia. Blood 100(5):1532-154 (2002).130'