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Primers, probes and kit for detecting specific methylation of colorectal tumor

A technology specific to colorectal tumors, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., to save medical resources, increase the detection rate of colonoscopy, and avoid the effect of colonoscopy reexamination

Pending Publication Date: 2019-10-25
阿吉安(福州)基因医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Primers, probes and kit for detecting specific methylation of colorectal tumor
  • Primers, probes and kit for detecting specific methylation of colorectal tumor
  • Primers, probes and kit for detecting specific methylation of colorectal tumor

Examples

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Embodiment 1

[0040] A kit for human colorectal tumor-specific methylation detection, specifically including the following components:

[0041] dNTP mixed solution, Mg 2+ Solution, DNA polymerase, PCR buffer, PCR grade deionized water, SDC2 / ACTB reaction solution, SFRP2 / ACTB reaction solution, negative control, positive control and blank control;

[0042] The SDC2 / ACTB reaction solution is composed of forward primer, reverse primer, probe of SDC2 gene and forward primer, reverse primer, probe of ACTB internal reference gene;

[0043]The SFRP2 / ACTB reaction solution is composed of forward primer, reverse primer, probe of SFRP2 gene and forward primer, reverse primer, probe of ACTB internal reference gene;

[0044] The negative control is: SDC2 gene, SFRP2 gene both show colorectal tumor and precancerous lesion-specific methylation-negative genomic sequence or synthetic gene sequence;

[0045] The positive control is the SDC2 gene, and the SFRP2 gene all presents genomic sequences or synthe...

Embodiment 2

[0051] 1. Sample selection

[0052] The stool samples of 1 patient diagnosed with colon cancer by colonoscopy and pathological examination were clinically collected, and the stool samples of 20 patients confirmed normal by colonoscopy examination were collected.

[0053] 2. DNA extraction from stool samples

[0054] DNA extraction was carried out on the obtained stool samples, using the stool DNA extraction kit of Ajian (Fuzhou) Gene Medicine Laboratory Co., Ltd., the specific steps are as follows:

[0055] 1) After fully dissolving all the samples, shake the stool specimen collection and storage tubes until they are evenly mixed, transfer 1 mL of the samples to a 15 mL centrifuge tube with a tipped 1 mL pipette tip, and mark the tubes;

[0056] 2) Add 1.5mL lysate to each tube, tighten the cap tightly, vortex and mix well, place in a dry bath at 70°C for 10min, during which time, gently invert and mix 3 times discontinuously, and cool to room temperature;

[0057] 3) Place ...

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Abstract

The invention discloses primers, probes and a kit for detecting the specific methylation of a colorectal tumor, and belongs to the technical field of biological detection. The primers and the probes include the forward primer, the backward primer and the probe of an SDC2 gene, the forward primer, the backward primer and the probe of an SFRP2 gene, and the forward primer, the backward primer and the probe of an ACTB reference gene. The kit comprises the primers and the probe. The primers, probes and kit for detecting the specific methylation of the colorectal tumor are designed to Chinese and have the advantages of high specificity and sensitivity, sampling is free of invasion and convenient since excrements are used as the sample, the ACTB is selected as the reference gene and is detectedin the same tube together with the SFRP2 gene and the SDC2 gene, the detection result is more accurate, and the primers, probes and kit have great significance in early finding precancerous lesions and early colorectal cancers.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a primer, a probe and a kit for detecting specific methylation of human colorectal tumors. Background technique [0002] Colorectal cancer (CRC) is a common malignant tumor in the digestive tract, including colon cancer and rectal cancer, also known as colorectal cancer, which is one of the major cancers that threaten human health. The most common clinical symptoms of colorectal cancer are blood in the stool, abdominal pain, abdominal distension, changes in bowel habits and stool properties. [0003] According to the national cancer statistics released by the National Cancer Center in 2018, there were 370,000 new cases of colorectal cancer in my country in 2014, and the incidence rate was 27.08 / 100,000, ranking third in the incidence of malignant tumors; the death rate of colorectal cancer was 180,000, with a mortality rate of 13.13 / 100,000, ranking fifth...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154C12Q2600/166
Inventor 袁青郭文浒薛怡婷陈浩段佳丽
Owner 阿吉安(福州)基因医学检验实验室有限公司
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