b'Gel & Capillary | IVD Assays FLT3 Mutation Assay - Gel DetectionGel and Capillary Assays andAssays FLT3 Mutation Assay - Gel DetectionGel and Capillary Assays andAssaysFLT3 Mutation Assay - Gel Detection ReagentsControls Concentration Units in 33 Reaction Assay Units in 330 Reaction AssayIVS-0050 Clonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubesIVS-P004 Clonal Control DNA 171 pg/mL 1 x 100 L tube 5 x 100 L tubesIVS-0000 Polyclonal Control DNA 200 g/mL 1 x 100 L tube 5 x 100 L tubesMaster Mixes Target Units in 33 Reaction Assay Units in 330 Assay MegaKitAssay DescriptionPerformance CharacteristicsFLT3 ITD Master Mix - Unlabeled FLT3 ITD 1 x 1500 L tube 10 x 1500 L tubesThe LeukoStrat FLT3 Mutation Assay is an in vitro diagnostic productThis LeukoStrat FLT3 Mutation Assay offers a rapid and reliableFLT3 ITD Master Mix - Unlabeled FLT3 TKD 1 x 1500 L tube 10 x 1500 L tubesintended for PCR-based detection of FLT3 activating mutations inmethod for detecting FLT3 mutations. This is shown by a validationSpecimen Control Size Ladder Multiple Genes 1 x 1500 L tube 10 x 1500 L tubespatients 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 TestGel Electrophoresis Detection LabPMM tested 57 blinded patient samples obtained from threeAgarose gel electrophoresis is used to resolve the different amplicon products based on their size, charge, AML in general has a poor prognosis. Assessment of the mutation statusindependent institutions. The institutions determined that 13 of theand conformation. Since DNA is negatively charged, when an electrical potential (voltage) is applied across of the FLT3 (fms related tyrosine kinase 3) receptor gene in karyotypesamples were FLT3 ITD positive, 33 were FLT3 ITD negative, 6 werethe gel containing PCR products, the electrical field causes the amplicons to migrate through the gel. Smaller normal AML is the most important prognostic indicator of diseaseFLT3 TKD positive, and 50 were FLT3 TKD negative. In addition 10DNA fragments are able to easily migrate through the gel matrix, whereas larger DNA fragments migrate outcome, which is often substantial, as many studies in AML havepositive blinded spiked samples and 10 negative samples were usedmore slowly. This causes a separation of the amplicon products based on size. Ethidium bromide or other shown that the presence of FLT3 activating mutations portends a poorfor the validation of FLT3 TKD. The LeukoStrat FLT3 Mutation AssayDNA intercalating dyes can then be used to stain and detect these products in the gel.prognosis. 1, 2For this reason, FLT3 activation mutation testing is requiredshowed a sensitivity and specificity of 100% with both master mixes. to stratify disease and determine appropriate treatment options. The analytical sensitivity of both master mixes was determined to be This PCR-based assay targets regions of the FLT3 gene to identify ITD5 positive cells out of 100 total cells.mutations and TKD mutations (such as the D835 and I836 mutations)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. Ordering InformationCatalog # Products Quantity9-412-0010 LeukoStrat FLT3 Mutation Assay - Gel Detection 33 reactions9-412-0020 LeukoStrat FLT3 Mutation Assay MegaKit - Gel Detection 330 reactionsThese are in vitro diagnostic products, and are not available for sale or use within North America.82Invivoscribe 2021|83'