Novel Strategies for Development of a Multiplexed Test with Expanded Content for Spinal Muscular Atrophy

SummaryThe overall goal of this project is to develop a cost effective multiplexed diagnostic test for spinal muscular atrophySMAto rapidly and accurately analyze multiple classes of disease causingdisease modifying DNA variantsand carrier status markersUltimatelythe proposed test will have a far reaching impact by expanding the identification and stratification of patients who may benefit from new treatmentsand by improving the detection of SMA carriersThe proposed test will detect an expanded panel of variants that are not assessed by conventional testingThe broader content is important because age of onsetdisease severity and progression correlations with the nature of the SMNmutationsand the compensatory and modifying effects of SMNand other genesThe expanded SMA test will not only extend the molecular testing to benefit more prospective parents and patients with atypical symptoms including adult onset casesbut will also address SMA testing needs of underserved populationsThe expanded SMA test panel will build upon Asuragen s AmplideXPCR CE SMNKitRUO and CEIVDand take advantage of a novel strategy Multiplexed andampEfficient Resolution of Genetic ElementsMERGEPCR to assesscopy number variantsCNVsof SMNand SMNpathogenic mutations in SMNdisease modifying mutationsandsilent carrier markersWe will also incorporate a novel gene tagged genotyping strategy to determine whether a mutation is in the SMNor SMNgeneThe specific aims areAimDesign and optimize individual assays for a unified multiplex MERGE PCR CE test to detect multiple classes of disease causingdisease modifying DNA variants and carrier markersAimDevelop algorithms to support assay signal deconvolution and enable accurate genotype classification of CNVSNVand INDEL signals from multiple fluorophore channelsAimDevelop and verify a single tube multiplex PCR based testIn Phase IIwe will advance the technology into a cGMP kit development processenable market ready software for automatic variant callingand develop controls and standards for an in vitro diagnostic productThe project will benefit from Asuragen s years of experience optimizing multiplexing PCR chemistries to develop and commercialize high performance diagnostic PCR CE assays Project NarrativeWe propose to develop a novel diagnostic test kit for spinal muscular atrophy that will support informedtreatment decisions for more patientsThe assay has a novel design approach that will enable the detection ofa broad panel of genetic variants and the format will make broadreliableand cost effective testing readilyaccessible through most clinical laboratories