Digital Melt Curve Analysis Platform for Longitudinal Cancer Patient Monitoring

AbstractBuilding on its commercially available Absolute Q digital PCR platform, during the 24-month Phase 2 SBIR project, Combinati will complete the development of the first Digital Melt Curve Analysis (DMCA) platform to the market to further advance the adoption of digital genomics for highly accurate, sensitive and reproducible nucleic acid quantification for longitudinal patient monitoring. To provide the “whole product” solution and prove the function, we will demonstrate the DMCA platform with Luminex’s 11- plex ESR1 (Estrogen Receptor 1) assay and conduct Beta testing at Dana Farber Cancer Institute: 1. Three beta instruments and the analysis software capable of digital melt curve analysis. 2. Complete dMCA validation internally with commercially available melt calibration kits. 3. Demonstrate andlt;0.1% Mutation Allele Frequency of 11 cell-free DNA targets using LuminexCorporation’s discrete melt assay. 4. Beta testing at Dana Farber Cancer InstituteNarrativeSince PCR was invented back in 1983, it has become the gold standard for applications requiring quantification of nucleic acids. The continuous evolution of the technology enables PCR to be more quantitative (qPCR), more accurate, precise and reproducible (digital PCR). In parallel, with the invention of melt curve analysis in 1997, it opens another dimension in melt temperatures for applications requiring simple and inexpensive genotyping, high degree qualitative multiplexing without sequencing, and assay optimization. Despite that there is a handful of dPCR platforms in the market, none of them supports Melt Curve Analysis. By combining melt curve analysis with digital PCR, Combinati strives to accelerate the adoption of digital genomics for all nucleic acid quantification needs in research and clinical markets.