Patent ID: 9075063
Filing Date: 2015-07-07
Classification: C12Q,G01N

Abstract:
1. A method for determination of presence or absence of cancer cells in a tissue obtained from a colon of a subject comprising the steps of: extracting DNA from the tissue; carrying out bisulfite treatment to the DNA obtained from the step of extracting; amplifying DNA of a promoter region of COL4A2 by using the DNA from the bisulfite treatment step as a template, a first primer comprising the sequence of SEQ ID NO: 15 and a second primer comprising the sequence of SEQ ID NO: 16; obtaining a peak from a methylated DNA fragment and a peak from a non-methylated DNA fragment of the promoter region of COL4A2 by analyzing the amplified DNA by mass spectrometry; calculating a methylation rate of CpG sites in the promoter region of COL4A2 from an area ratio between a peak from the methylated DNA fragment and a peak from the non-methylated DNA fragment; and determining presence or absence of cancer cells in the tissue based on a comparison of the rate obtained from the step of calculating to a pre-determined cut-off value, and cancer cells are determined to be present in the tissue when the rate is higher than the cut-off value.