Polygene combined methylation detection method and application thereof

Abstract

The invention provides a polygene combined methylation detection method and application thereof, and the method comprises the following steps: step 1): carrying out first PCR amplification on a DNA sample treated by hydrosulfite to obtain a target template DNA fragment; 2) carrying out PCR amplification on the target template DNA fragment obtained in the step 1) for the second time, and carrying out biotin labeling on a methylation sequence in the DNA fragment; 3) hybridizing the PCR product labeled by biotin in the step 2) with a label complementary sequence on the hybridized magnetic bead, and carrying out fluorescence labeling by using a fluorescent dye; and 4) detecting the fluorescence labeled product obtained in the step 3), and judging the methylation condition of the target gene in the DNA sample. A plurality of methylated DNA samples can be amplified, a PCR product is hybridized with different magnetic bead complementary tag sequences, the gene methylation condition is determined through a fluorescence labeling product, and the whole detection process is rapid and high in accuracy; and the detection method is widely applied to early screening of canceration genes of diseases such as colon straight cancer, bladder cancer and cervical cancer.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a multi-gene joint methylation detection method and its application. Background technique [0002] Methylation is a modification method of gene epigenetics, which participates in the regulation of gene expression, and then participates in the process of cell growth and development, adversity stress response and so on. More and more studies have shown that abnormal DNA methylation and changes in DNA methylation patterns are closely related to the occurrence and occurrence of cancer. DNA methylation has been regarded as a promising molecular biomarker of cancer, which provides new insights for early diagnosis of cancer, assessment of cancer risk, formulation of treatment plan and prognosis judgment. [0003] Methylation occurs mainly at CpG dinucleotides in the human genome. The DNA methylation process is catalyzed by DNA methyltransferase, the methyl group on S-aden...

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to provide a multi-gene combined methylation detection method and its application, aiming at different genes and using primers to carry out PCR reaction, hybridizing the amplified PCR product with the label complementary sequence on the magnetic beads, through Fluorescent markers are used to detect the methylation of multiple gene sites. In this technical solution, two PCR amplifications can be used to amplify the methylation signal, thereby solving the problem of low sensitivity and accuracy in the existing gene methylation detection process. low technical issues

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