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Colorectal Cancer Molecular Marker Combination, Its Application, Primer Set and Detection Kit

A molecular marker, colorectal cancer technology, applied in the field of molecular biology, can solve the problem of low sensitivity, achieve high patient dependence, improve patient survival rate, and improve the effect of early diagnosis rate

Active Publication Date: 2020-12-01
SHENZHEN HAPLOX BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Septin9 gene detection is aimed at the detection of the methylation status of Septin9 gene in peripheral blood plasma. It is non-invasive, but its detection sensitivity is low, only 68%-79.3%.

Method used

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  • Colorectal Cancer Molecular Marker Combination, Its Application, Primer Set and Detection Kit
  • Colorectal Cancer Molecular Marker Combination, Its Application, Primer Set and Detection Kit
  • Colorectal Cancer Molecular Marker Combination, Its Application, Primer Set and Detection Kit

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Embodiment

[0052] In this embodiment, 17 subjects were selected, among which 11 were colorectal cancer stage I-II patients (CRC01-CRC11) and 6 were healthy people (H01-H06). Take peripheral blood samples from all people for testing. The specific testing steps and results are as follows:

[0053] (1) Separation of plasma samples

[0054] Peripheral blood samples were placed in 10 mL centrifuge tubes and centrifuged at 1,600 g for 10 min at 4°C. The supernatant was taken out, placed in a new 10 mL centrifuge tube, and centrifuged at 16,000 g for 10 min at 4°C. Remove the supernatant and place it in a new 10mL centrifuge tube.

[0055] (2) Extraction of cfDNA

[0056] Circulating nucleic acid (cfDNA) in plasma was extracted using Qiagen Circulating Nucleic Acid Kit according to the manufacturer's instructions.

[0057] (3) Bisulfite conversion

[0058] Take 20 ng of cfDNA, use the ZYMO EZ DNA Methylation Gold Kit, and convert the cfDNA to bisulfite according to the manufacturer's instr...

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Abstract

The invention provides a colorectal cancer molecular marker combination. The colorectal cancer molecular marker combination comprises at least two selected from the group consisting of an AKT1 gene shown as SEQ ID NO: 1 in a sequence table or a reverse complementary sequence thereof, an AXIN2 gene as shown in SEQ ID NO: 2 or a reverse complementary sequence thereof and a GNA12 gene as shown in SEQID NO: 3 or a reverse complementary sequence thereof, wherein the promoter regions of the AKT1 gene, the AXIN2 gene and the GNA12 gene are in a low methylation state in colorectal cancer people. Theinvention also provides an application of the colorectal cancer molecular marker combination in preparation of a reagent used for detecting the colorectal cancer, and a primer group and a test kit used for detecting the colorectal cancer. The colorectal cancer detection sensitivity of the colorectal cancer molecular marker combination reaches 90% or above, and even can reach 100%.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and relates to molecular markers for disease diagnosis, in particular to a colorectal cancer molecular marker combination, the use of the molecular marker combination in preparing reagents for detecting colorectal cancer, and the corresponding Primer sets and detection kits. Background technique [0002] Colorectal cancer is the third most common cancer in the world, with 1.4 million cases diagnosed and 700,000 deaths in 2012. Colorectal cancer is also one of the most common malignant tumors in my country. The 5-year survival rate of patients with early colorectal cancer is more than 90%, while the survival rate of patients with advanced colorectal cancer is only about 10%. However, since colorectal cancer often has no special symptoms in the early stage, it is easy to delay the timing of treatment, and about 60% of the patients are in the middle and late stages when they are diagnosed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 许明炎张晓妮屈宏越
Owner SHENZHEN HAPLOX BIOTECH
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