b'Gel & Capillary | IVD Assays FLT3 Mutation Assay 2.0Gel and Capillary Assays andAssaysABI Fluorescence Detection FLT3 Mutation Assay 2.0 -ABI Fluorescence DetectionAssay Description Table 1. FLT3 ITD Percent Agreement with 454 SequencingThe LeukoStrat FLT3 Mutation Assay 2.0 is an in vitro diagnosticPercent Agreement Discordance # Concordance # *95% LLproduct intended for PCR-based detection of FLT3 activating mutations in patients with acute myelogenous leukemia (AML). Negative PA 100% 0 119 96.9%Specifically, the FLT3 Mutation Assay 2.0 can be used to: Positive PA 98.0% 4 200 95.1%Identify internal tandem duplications (ITD) in the FLT3 gene * 95% of results would be expected to agree with sequencing at a rate greater than or equal to the lower limit (LL).Identify tyrosine kinase domain (TKD) mutations in the FLT3 geneSummary and Explanation of the TestTable 2. FLT3 TKD Percent Agreement with 454 SequencingAML in general has a poor prognosis. 1,2Many studies in AML have shown that the presence of FLT3 (fms related tyrosine kinase 3)Percent Agreement Discordance # Concordance # *95% LLactivating mutations portends a poor prognosis making it an attractive target for treatment. For this reason, FLT3 mutation testing is required toNegative PA 100% 0 137 96.9%stratify disease and determine appropriate treatment options.Positive PA 100% 0 240 98.5%Using this assay, DNA is amplified via PCR with fluorophore labeled*95% of results would be expected to agree with sequencing at a rate greater thanprimers, TKD amplicon is enzymatically digested, and FLT3 mutationsor equal to the lower limit (LL).are detected via capillary electrophoresis. This test kit includes 2 PCR master mixes, along with positive and negative controls for mutant detection. Each master mix (FLT3 ITD Master Mix and FLT3 D835 Master Mix) contains a fluorophore-labeled PCR primer set for the respectiveReferencesdetection of internal tandem duplication or TKD mutation. 1. Murphy KM et al., A Clinical PCR/Capillary Electrophoresis Assay for the Detection of Internal Tandem Duplication and Point Mutation of Performance Characteristicsthe FLT3 Gene. J. Mol. Diag. 5:96-102 (2003).This assay can reliably detect mutations comprising more than 5% of2. Yamamoto, Y., et al., Activating mutation of D835 within the the total cell population. Also, as demonstrated herein, the LeukoStratactivation loop of FLT3 in human hematologic malignancies. Blood, FLT3 Mutation Assay 2.0 detects FLT3 ITD and TKD mutations with97(8):2434-9 (2001).excellent concordance to NGS methodologies (Table 1, Table 2).3.Acute Myeloid Leukemia, Clinical Practice Guidelines in Oncology, National Comprehensive Cancer Network (v.2.2014)86'