Ratiometric Quantification of Targeted Methylated DNA sites using MM-LAMP-OSD & Detection Devices

The DoD is seeking technology and systems to quantify regulating DNA methylation transcriptional activators or repressors, and chromatin remodeling enzymes. Fabrico Technology, Dr. Sanchita Bhadra, and Professor Andrew Ellington of the University of Texas at Austin, proposes to develop an in-field detection platform based upon Mismatch Methyl Loop Mediated Isothermal Amplification-One-step Strand Displacement (MM-LAMP-OSD), a method that should prove capable of the quantitative identification of methylated nucleotides at targeted sites. MM-LAMP-OSD is dependent upon the innovation that antibodies can specifically recognize methylated nucleotides where a mismatch occurs resulting in a steric block to isothermal amplification via LAMP. Fabrico Technology has developed a patent pending LAMP-OSD detection system which is ideally suited for determining quantitatively this MM-LAMP-OSD assay ratiometrically. In this proposal effort, Fabrico and the Ellington Lab intends to modify and optimize the device to ratiometrically measure multiple MM-LAMP-OSD assays. By ratiometrically comparing non-fluorescent methylated DNA sequences to the fluorescence of non-methylated DNA sequences, it possible to quantitatively determine methylation sites in nucleic acid bases from a mammalian genome resulting in a novel method for monitoring any stage of a disease, onset, clinical course, or response to treatment.