b'AML -MRD TestsFLT3 ITD MRD AssayClinical Information Test Name Indications for TestingMinimal residual disease (MRD) detection in patientsThe treatment of AML has become a paradigm forFLT3 ITD MRD Assay dentify tumor-specific markers for post-treatment Iwith leukemia has proven to be useful in the clinicalprecision medicine. This MRD assay is at least two ordersmonitoring management of disease and can facilitate the developmentof magnitude more sensitive than other commerciallyAssay Type Monitor and evaluate disease recurrenceof new therapies. Mutations in the fms-like tyrosine kinaseavailable FLT3 assays. It detects the persistence of a driver 3 (FLT3) gene are the most prevalent mutations found inmutation, FLT3 ITD, in patients with no overt evidenceNext-Generation Sequencing (NGS)acute myeloid leukemia (AML) 1and are characterized byof disease, allowing clinicians to identify those patientsCLIA-validated assayan aggressive phenotype with a high prevalence of relapse.that can benefit from continuation or modification of Internal tandem duplication (ITD) mutations within thetreatment. 3juxtamembrane domain are the most common mutationsMethod Descriptionof FLT3. 2The development of a sensitive and specificMRD detection by Next-Generation Sequencing hasTo track and identify previously detected FLT3 ITD mutations assay for FLT3 ITD mutations represents a significantdemonstrated utility in predicting clinical outcomesin post-treatment follow-up samples, a multiplex master advancement in guiding treatment decisions. and in generating clinically actionable results, allowingmix targeting the juxtamembrane domain of the FLT3 gene LabPMMs FLT3 ITD MRD test is an NGS-based, targeted,early intervention, confirmation of disease status prior tois used to amplify DNA extracted from a patient sample.deep-sequencing assay that detects ITDs ranging from 3transplant, and increased confidence in remission status.bp to over 200 bp in size. Once a specific ITD (length andNext-generation sequencing of the PCR products is used sequence) has been identified in a primary sample, it canto identify DNA sequences specific to previously identified easily be tracked in subsequent samples at a sensitivity mutations detected at diagnosis. Bioinformatics tools facilitate the detection of these specific sequences present of 5x10 -5 , provided sufficient DNA quantity is tested. at an allelic sensitivity level of 5x10 -5 .Interpretation TurnaroundSpecimenShippingSpecimen Time Requirements Conditions StabilityAn interpretive7 to 10 1-3 mL of peripheral Ambient or Cool; 2-8 C up report will bebusiness days blood in EDTA,do not freezeto 7 days priorissued indicating0.25-1 mL of bone marrow (peripheral blood orto testing whether FLT3 ITDbone marrow) MRD was detected in EDTA 1 g of previously Ambient or frozen on isolated DNAdry ice (isolated DNA) References1.The Cancer Genome Atlas Research Network (2013) Genomic and Epigenomic Landscapes of Adult De Novo Acute MyeloidLeukemia. N Engl J Med. 368: 20592074.2.Konig H. et al. (2015) Targeting FLT3 to treat leukemia. Expert Opin Ther Targets 19:37-54.3.Levis, M. J. et al (2018) A next-generation sequencingbased assay for minimal residual disease assessment in AML patientswith FLT3-ITD mutations. Blood Advances, 2: 825-83136 LabPMM Services Catalog 2021|37'