DNA integrity assay (DIA) for cancer diagnostics, using confocal fluorescence spectroscopy

摘要

The present invention relates, e.g., to a method for determining the size distribution of DNA molecules in a sample comprising cell-free nucleic acid, comprising labeling the DNA with a fluorescent dye in a stoichiometric manner, subjecting the DNA to molecular spectroscopy (e.g., cylindrical illumination confocal spectroscopy), analyzing suitable fluorescent burst parameters of the labeled DNA, and conducting single molecule DNA integrity analysis of the labeled DNA molecules in the sample. In one embodiment of the invention, the method is used as a diagnostic method for detecting cancer.

主权项

1. A method for determining the size distribution of cell-free DNA molecules in a sample, comprising
labeling the cell-free DNA molecules in the sample with a fluorescent dye in a stoichiometric manner, subjecting the labeled DNA to single molecule spectroscopy, and measuring fluorescent burst parameters and fluorescent burst rate of the labeled DNA, wherein the fluorescent burst parameters, which comprise burst size, burst height and/or transit time, indicate the length of each of the DNA molecules; and the fluorescent burst rate indicates the abundance of each size of DNA molecule, and conducting single molecule DNA integrity analysis (smDIA) of the labeled DNA molecules in the sample, thereby determining the size distribution of the cell-free DNA molecules.