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Primer and kit based on human colorectal cancer specific methylation detection

A colorectal cancer and methylation technology, applied in the field of disease diagnosis and prevention, disease diagnosis and detection, can solve the problems of high false positive rate, cumbersome operation, high cost, etc., and achieve the effect of sensitivity and specificity

Active Publication Date: 2014-11-26
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have disadvantages such as cumbersome operation, high false positive rate, and high cost, which limit their wide application in clinical laboratories.

Method used

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  • Primer and kit based on human colorectal cancer specific methylation detection
  • Primer and kit based on human colorectal cancer specific methylation detection
  • Primer and kit based on human colorectal cancer specific methylation detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 A kind of specific primer design for human colon cancer detection

[0034] The PCR primers for human colon cancer-specific methylation of the present invention are designed according to the human whole genome sequence disclosed by NCBI (National Center for Biotechnology Information), using Primer Premier3. Synthetic Industry Co., Ltd.

[0035] Forward primer: 5-CGTAGTTGGATGGGATTATTT-3 (SEPTIN9-Forward, SEQ ID NO.1)

[0036] Reverse primer: 5-GCCAAACCCACCCGCAAA-3 (SEPTIN9-Reverse, SEQ ID NO.2)

Embodiment 2

[0037] Example 2 A kit for methylation detection of human colon cancer

[0038] Including the following primer sequences and PCR system:

[0039]Forward primer: 5-CGTAGTTGGATGGGATTATTT-3 (SEPTIN9-Forward, SEQ ID NO.1)

[0040] Reverse primer: 5-GCCAAACCCACCCGCAAA-3 (SEPTIN9-Reverse, SEQ ID NO.2)

[0041] Also included in the described test kit: 1×Master, 25mM MgCl 2 , methylated standard DNA and unmethylated standard DNA.

Embodiment 3

[0042] Example 3 Application of the kit of the present invention in the diagnosis of human colon cancer

[0043] The Master used in the present invention was purchased from Roche Co., Ltd.; the DNA extraction kit was QIAmp DNA Mini Kit (QIAGEN); the transformation kit was EZ DNA Methylation kit (Zymo Research, Orange, CA, USA); other reagents were domestic Analytical reagents.

[0044] The biological materials used in the present invention are all from Shanghai Ruijin Hospital.

[0045] method:

[0046] 1. Biological samples:

[0047] The tumor tissues and normal tissues of 33 patients with colorectal cancer who were treated in Shanghai Ruijin Hospital from March to September 2013 were collected.

[0048] 2. Extract tissue DNA:

[0049] 1. Take about 25mg of colorectal cancer tissue or normal tissue, chop it up with surgical scissors after autoclaving and put it into a 1.5ml centrifuge tube, then add 180μl of Buffer ATL;

[0050] 2. Take 20 μl of QIAGEN Protease and add i...

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Abstract

The invention discloses a primer and system for human colorectal cancer specific methylation detection as well as an application thereof. A pair of PCR primers and a kit (a system) used for human colorectal cancer specific methylation detection are designed in a promoter area aiming at SEPTIN9 genes in a human colorectal cancer genome. A sample to be detected and vulcanized is subjected to PCR detection by using the primers and system, and whether the sample is a human colorectal cancer sample is judged according to the relative fluorescence intensity after amplification. Compared with a conventional colorectal cancer diagnosis method, the method disclosed by the invention has the characteristics of rapidness, high sensitivity, high specificity and the like and can be used for timely diagnosing the human colorectal cancer at early stage.

Description

technical field [0001] The invention relates to the field of disease diagnosis and detection, in particular to a primer, system and application for colorectal cancer-specific methylation detection, belonging to the field of disease diagnosis and prevention. Background technique [0002] Colorectal cancer is a malignant tumor that seriously threatens human health, and its incidence is second only to lung cancer and breast cancer, ranking third among malignant tumors. WHO statistics show that the 5-year survival rate of patients with colorectal cancer is 61% in North America, but only 32% in my country. According to the study, the 5-year survival rate is significantly correlated with the severity of the disease at the time of diagnosis. The 5-year survival rate of patients with advanced stage is less than 10%, while the 5-year survival rate of patients with early-diagnosed colorectal cancer can reach 92%. %. From 2008 to 2013, the incidence and mortality of colorectal cancer...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 张宏莲毛红菊郭慧卫旭宇金庆辉赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI