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A colorectal cancer screening marker composition, its selection method, and colorectal cancer screening kit

A colorectal cancer and kit technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/testing, etc., can solve the problems of unacceptable colonoscopy and low detection sensitivity of early lesions, and achieve improved inspection Sensitivity, increased acceptance, and reduced mortality effects

Active Publication Date: 2022-02-22
南京求臻基因科技有限公司
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Problems solved by technology

[0006] The main purpose of the present invention is to provide a colorectal cancer screening marker composition, which aims to solve the problems of low detection sensitivity of the existing colorectal cancer diagnosis methods for early lesions and unacceptable colonoscopy

Method used

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  • A colorectal cancer screening marker composition, its selection method, and colorectal cancer screening kit
  • A colorectal cancer screening marker composition, its selection method, and colorectal cancer screening kit
  • A colorectal cancer screening marker composition, its selection method, and colorectal cancer screening kit

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Embodiment

[0086] 1. Take 5 mL of plasma and use Qiagen Plasma Free DNA Extraction Kit (Cat: 55204) to extract cfDNA.

[0087] 2. Take 20 ng of extracted cfDNA and HinP1I (Cat: R0124, NEB), HpaII (Cat: R0171, NEB), AciI (Cat: R0551, NEB), HpyCH4IV (Cat: R0619, NEB) four methylation sensitive Endonuclease (final concentration 10 U / μL), 20 μL system, incubate at 37°C for 16 h, inactivate the enzyme at 80°C for 20 min, the enzyme digestion system is shown in Table 3, cfDNA from leukocytes is cut.

[0088] Table 3 Methylation-sensitive endonuclease digestion system

[0089]

[0090] 3. Use all the incubated products as templates, add the above 6 pairs of target-specific primers (50 nM each) and 1 pair of internal reference primers (10 nM), and configure the system for multiplex PCR. The reaction system is shown in Table 4. Multiplex PCR The reaction program is: 98°C / 45 s; 8cycles (98°C / 15 s, 55°C / 30 s, 72°C / 30 s); 72°C / 1 min; 4°C / hold, multiple PCR products are obtained from the reaction...

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Abstract

The present invention proposes a colorectal cancer screening marker composition, its selection method, and a colorectal cancer screening kit. The colorectal cancer screening marker composition is hypermethylated in colorectal cancer and hypomethylated in white blood cells , and has a methylation-sensitive endonuclease cleavage site, which can degrade non-tumor-derived cfDNA and specifically amplify tumor-derived ctDNA by using the property that methylation-sensitive endonuclease cannot cut methylated sites, greatly improving the inspection sensitivity. Moreover, the detection of ctDNA is non-invasive, fast, convenient, and non-invasive, which can improve patients' acceptance of colorectal cancer screening. Through more accurate and effective detection and screening of early colorectal cancer, the mortality rate of colorectal cancer and the harm of colorectal cancer can be reduced.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a colorectal cancer screening marker composition, a selection method thereof, and a colorectal cancer screening kit. Background technique [0002] Colorectal cancer (CRC) is the fourth most common cancer in men and the second most common cancer in women in the world, with the highest incidence in developed countries. In recent years, with the improvement of people's living standards in our country, the change of living habits and dietary structure, and the increasingly serious environmental pollution, the incidence of colorectal cancer in our country has been increasing. According to the 2013 research report of the National Cancer Early Diagnosis and Early Treatment Project Expert Committee, the rate of advanced adenoma and early bowel cancer among high-risk groups over the age of 40 in my country is 6%. According to the 2015 "Annual Report on Cancer Registration in C...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/154C12Q2600/166C12Q2600/16C12Q2531/113C12Q2521/301C12Q2545/101C12Q2537/143C12Q2563/107
Inventor 段小红张怡然王冰杨春燕邵茗泽王天阳
Owner 南京求臻基因科技有限公司
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