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Kit for detecting DNA methylation markers of early colorectal cancer and adenoma

A technology for methylation markers and colorectal cancer, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial measurement/testing, etc., can solve the problems of high technical requirements for examiners and high missed detection rate of endoscopic screening

Pending Publication Date: 2021-11-26
SHANGHAI GENECHEM CLINICAL LAB INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Intestinal adenoma can be screened by endoscopy, but the technical requirements for examiners are relatively high, and the missed detection rate of clinical endoscopy screening is high

Method used

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  • Kit for detecting DNA methylation markers of early colorectal cancer and adenoma
  • Kit for detecting DNA methylation markers of early colorectal cancer and adenoma
  • Kit for detecting DNA methylation markers of early colorectal cancer and adenoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0147] Example 1 TaqMan-PCR method for detection of methylation sites in stool samples

[0148] 1.1 Extraction and transformation of DNA samples

[0149] Human NDRG4 and ACTIN gene fragments were extracted from fecal samples using magnetic beads linked to specific probes, eliminating the interference of microorganisms and other human DNA. Among them, NDRG4 is used as the detection target, and ACTIN is used as the internal reference.

[0150] The sample used in this example is feces, which is convenient for sampling. The preservation of feces samples uses the prepared sample preservation solution, which can preserve the samples at room temperature for up to 7 days, which reduces the difficulty of sample transportation. Among them, the sequence of the specific probe linked on the magnetic beads is as follows:

[0151]

[0152] Sample Preservation Solution Formula:

[0153] ingredients Sodium acetate Sodium chloride EDTA SDS concentration 0.1M 0.5M...

Embodiment 2

[0217] Example 2 MS-HRM method for detection of methylation sites in tissue samples

[0218] Another method for detecting methylation sites in a sample—methylation-sensitive high-resolution melting curve analysis (MS-High Resolution Melting Curve Analysis) is also provided in this embodiment. The MS-HRM method is a simple and sensitive method for detecting gene methylation levels that does not require subsequent manipulation of PCR products. This method is mainly realized by comparing the melting temperature and peak shape of the curve and can detect a single methylation in a series of CpG sites, and at the same time, it can also detect the methylation of a series of CpG sites level to analyze. The methylation of individual CpG sites and the average methylation level will affect the shape of the melting curve. Before the PCR reaction, design primers at non-CpG island sites, so that the middle of the pair of primers contains meaningful methylated CpG islands. Once these CpG i...

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Abstract

The invention provides a DNA methylation marker for diagnosis, screening and risk prediction of early colorectal cancer and adenoma. The marker is characterized in that a CpG site in an NDRG4 gene is simultaneously methylated at least two sites of the 38th site, the 47th site, the 50th site and the 52th site in SEQ ID NO.:1. The invention also provides a method, a primer pair, a probe and a kit for detecting the DNA methylation marker. The method provided by the invention has good specificity and high sensitivity on early colorectal cancer and adenoma, is low in detection cost and simple to operate, and is beneficial to wide application of early colorectal cancer screening.

Description

[0001] This application is an invention patent with an application date of July 29, 2020, an application number of 202010745610.X, and an invention title of "DNA methylation markers for early colorectal cancer and adenoma, methods for detecting them and their applications" Divisional application of the application. technical field [0002] The invention relates to the field of gene detection, in particular, the invention relates to a methylation marker of early colorectal cancer and adenoma, a detection method and application thereof. Background technique [0003] In China, the incidence of colorectal cancer ranks fourth among all malignant tumors. Insufficient screening of colorectal cancer in China is the main reason for the low five-year survival rate of colorectal cancer in my country. The latest research shows that feces contain a small amount of nucleic acids derived from human intestinal epidermal exfoliated cells, and multi-target detection of these nucleic acids ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/6886C12Q1/6883C12Q1/686C12Q2600/154C12Q2600/112C12Q2561/101C12Q2563/107
Inventor 何逖吴帅来姚浩
Owner SHANGHAI GENECHEM CLINICAL LAB INC
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