b'Gel & Capillary | IVD Assays FLT3 Mutation Assay - Gel DetectionGel and Capillary Assays andAssaysFLT3 Mutation Assay - Gel DetectionAssay DescriptionPerformance Characteristics The LeukoStrat FLT3 Mutation Assay is an in vitro diagnostic productThis LeukoStrat FLT3 Mutation Assay offers a rapid and reliable intended for PCR-based detection of FLT3 activating mutations inmethod for detecting FLT3 mutations. This is shown by a validation patients with acute myelogenous leukemia (AML). performed by the Laboratory for Personalized Molecular Medicine (LabPMM). Specifically, the FLT3 Mutation Assay can be used to:Identify internal tandem duplications (ITD) in the FLT3 gene Concordance with three independent labsIdentify tyrosine kinase domain (TKD) mutations in the FLT3 gene ITD 100% sensitivity / 100% specificity TKD 100% sensitivity / 100% specificitySummary and Explanation of the Test AML in general has a poor prognosis. Assessment of the mutation statusLabPMM tested 57 blinded patient samples obtained from three of the FLT3 (fms related tyrosine kinase 3) receptor gene in karyotypeindependent institutions. The institutions determined that 13 of the normal AML is the most important prognostic indicator of diseasesamples were FLT3 ITD positive, 33 were FLT3 ITD negative, 6 were outcome, which is often substantial, as many studies in AML haveFLT3 TKD positive, and 50 were FLT3 TKD negative. In addition 10 shown that the presence of FLT3 activating mutations portends a poorpositive blinded spiked samples and 10 negative samples were used prognosis. 1, 2For this reason, FLT3 activation mutation testing is requiredfor the validation of FLT3 TKD. The LeukoStrat FLT3 Mutation Assay to stratify disease and determine appropriate treatment options. showed a sensitivity and specificity of 100% with both master mixes. This PCR-based assay targets regions of the FLT3 gene to identify ITDThe analytical sensitivity of both master mixes was determined to be mutations and TKD mutations (such as the D835 and I836 mutations) 5 positive cells out of 100 total cells.in sample human genomic DNA. DNA is amplified by PCR, TKD amplicon is enzymatically digested, and FLT3 mutations are detected via gel electrophoresis. References This test kit includes 3 master mixes, along with positive and negative1. Murphy KM et al., A Clinical PCR/Capillary Electrophoresis Assay for controls for mutant detection. (1) FLT3 ITD Master mix tests for internalthe Detection of Internal Tandem Duplication and Point Mutation of tandem duplication mutations. (2) FLT3 D835 Master Mix tests forthe FLT3 Gene. J. Mol. Diag. 5:96-102 (2003).TKD region mutations. (3) The Specimen Control Size Ladder master2. Yamamoto, Y., et al., Activating mutation of D835 within the mix targets multiple genes and generates a series of amplicons ofactivation loop of FLT3 in human hematologic malignancies. Blood, approximately 100, 200, 300, 400, and 600 basepairs to ensure that the97(8):2434-9 (2001).quality and quantity of input DNA is adequate to yield a valid result.84'