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Kit for identification of gastric cancer and/or gastric polyps and application thereof

A kit and technology for gastric polyps, applied in the biological field, can solve problems such as pain, improve sensitivity and specificity, and ensure correctness and reliability.

Inactive Publication Date: 2018-12-11
BEIJING EXELLON MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the process of gastroscopy is very painful, and less than 10% of patients can really do regular endoscopy
However, there is still a lack of means to effectively detect the methylation status of these cancer-related genes and process the detection results

Method used

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  • Kit for identification of gastric cancer and/or gastric polyps and application thereof
  • Kit for identification of gastric cancer and/or gastric polyps and application thereof
  • Kit for identification of gastric cancer and/or gastric polyps and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1: DNA Extraction

[0061] DNA Extraction Reagent consists of Lysis Buffer, Binding Buffer, Washing Buffer and Elution Buffer. Lysis buffer consists of protein denaturants, detergents, pH buffers and nuclease inhibitors. Binding buffers consist of protein denaturants and pH buffers. The cleaning buffer is divided into cleaning buffer A and cleaning buffer B, and cleaning buffer A is composed of protein denaturant, nuclease inhibitor, detergent, pH buffer and ethanol; cleaning buffer B is composed of nuclease inhibitor, pH buffer and ethanol composition. The elution buffer consists of nuclease inhibitors and pH buffers. The protein denaturant is: guanidine hydrochloride; the detergent is: Tween20; the pH buffer is: Tris-HCl; the nuclease inhibitor is: EDTA.

[0062] In this embodiment, the plasma samples of patients with gastric cancer are taken as an example to extract plasma DNA. The extraction method comprises the following steps:

[0063] (1) Take 1ml o...

Embodiment 2

[0075]Example 2: Bisulfite Treatment of DNA

[0076] Bisulfite treatment of DNA is treated with bisulfite reagent, which is composed of bisulfite buffer and protection buffer; bisulfite buffer is a mixed liquid of sodium bisulfite and water ; The protection buffer is a mixed liquid of oxygen free radical scavenger hydroquinone and water.

[0077] In this embodiment, the DNA extracted in Example 1 is used as the processing object, and the DNA is treated with bisulfite, and the specific steps include:

[0078] (1) Preparation of bisulfite buffer: weigh 1g of sodium bisulfite powder, add water to prepare 3M buffer;

[0079] (2) Preparation of protection buffer: weigh 1g of hydroquinone reagent, add water to prepare 0.5M protection buffer;

[0080] (3) Mix 100 μl DNA solution, 200 μl bisulfite buffer solution and 50 μl protection solution, shake and mix evenly;

[0081] (4) Thermal cycle: 95°C for 5 minutes, 80°C for 60 minutes, 4°C for 10 minutes;

[0082] (5) Add 1ml DNA bin...

Embodiment 3

[0092] Example 3: Detection of DNA methylation by real-time fluorescent PCR and verification of primer sets

[0093] In this embodiment, real-time fluorescent PCR is used as an example to measure the methylation level of biomarker genes. The detected genes were CDH1, DAPK, PAX5, RASSF1A, Reprimo, RNF180, RUNX3, SDC2, Septin9 and TCF4 genes, and the internal reference gene was ACTB. In this example, the DNA treated with bisulfite in Example 2 was used as a template for real-time fluorescent PCR amplification. The DNA samples to be tested, the negative quality control, the positive quality control, and the no-template control were all tested in triplicate holes.

[0094] For CDH1, DAPK, PAX5, RASSF1A, Reprimo, RNF180, RUNX3, SDC2, Septin9 and TCF4 genes, many sets of primer and probe combinations can be designed, and the performance of each set of probe primer combinations may be different, so it is necessary to pass Experiment to verify.

[0095] Therefore, the present inven...

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Abstract

The invention relates to a kit for the identification of gastric cancer and / or gastric polyps and application thereof. The kit comprises a primer pair, wherein the primer pair is used for detecting the methylation level of a biological marker gene or corresponding segments in a biological sample of a tester; the primer pair is used for performing PCR (polymerase chain reaction) amplification reaction by using the biological marker gene or segments thereof treated by bisulfite as a template; the biological marker gene is selected from one or multiple of CDH1, DAPK, PAX5, RASSF1A, Reprimo, RNF180, RUNX3, SDC2, Septin9 and TCF4. The kit has the advantage that by jointly detecting the methylation level of the biological marker gene or corresponding segments, the sensitivity and specificity indetection of the gastric cancer and / or gastric polyps is improved, so that the correctness and reliability of the detection results are guaranteed, and a quick, reliable and accurate novel path is provided for the predicting, diagnosis and evaluation on the gastric cancer and / or gastric polyps.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for identifying gastric cancer and / or gastric polyps and application thereof. Background technique [0002] Gastric cancer is one of the most common malignant tumors worldwide. According to the data provided by WHO: there are more than 1.5 million gastric cancer patients worldwide, and its mortality rate ranks third among cancer deaths worldwide. Gastric cancer occurs most commonly in East Asia and Eastern Europe. China accounts for more than 40% of the new gastric cancer cases and related deaths in the world, ranking first in the world. Adding Japan and South Korea, the gastric cancer rate accounts for about 2 / 3 of the global patients. The International Agency for Research on Cancer (IARC), a subsidiary of WHO, speculates that the high incidence of gastric cancer in the three Northeast Asian countries may be related to race (genetics), eating habits, and aging. In terms of...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6883
CPCC12Q1/6883C12Q1/6886C12Q2600/154C12Q2600/166
Inventor 李淑钰蒲珏李明明徐春叶陈彦利
Owner BEIJING EXELLON MEDICAL TECH CO LTD
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