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Composition for detecting gastric cancer as well as kit and use thereof

The technology of a composition and a kit, which is applied in the field of compositions for detecting gastric cancer, can solve the problems of low diagnosis rate and high detection cost, and achieve the effect of effective detection and good detection sensitivity

Active Publication Date: 2018-06-29
BIOCHAIN BEIJING SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problems of low diagnosis rate and high detection cost in the detection of early gastric cancer patients in the prior art, the present invention provides a composition for detecting gastric cancer. The composition provided by the present invention can detect gastric cancer sensitively and specifically. The present invention also provides a kit containing the composition and its use in the detection of gastric cancer. The kit provided by the present invention has good detection sensitivity for gastric cancer and can detect gastric cancer conveniently, quickly and effectively

Method used

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  • Composition for detecting gastric cancer as well as kit and use thereof
  • Composition for detecting gastric cancer as well as kit and use thereof
  • Composition for detecting gastric cancer as well as kit and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Example 1 Detection of methylation status of SEPT9 gene and RNF180 gene

[0132]Plasma samples from 35 patients with gastric cancer and 67 normal subjects were obtained. All samples come from Boercheng Company. Then the genomic DNA of gastric cancer and normal people is extracted, and the genomic DNA samples are pretreated so that the unmethylated cytosine base at the 5' position is converted into uracil, thymine or not used in the hybridization behavior. Another base for pyrimidine. In this example, this pretreatment was achieved by bisulfite reagent treatment. The extraction and processing of the DNA can be carried out by any standard means in the prior art. Specifically, in this embodiment, all sample DNA extraction and bisulfite DNA modification are carried out by using Boercheng Company Extracted from plasma processing kit. Then, the aforementioned SEPT9 gene and RNF180 gene primers, probe combinations and internal reference gene ACTB gene primers and probe c...

Embodiment 2

[0135] The ELISA detection of embodiment two MG7 antigen

[0136] Peripheral blood samples from 35 patients with gastric cancer and 67 normal subjects were obtained. All samples come from Boercheng Company. First, the serum concentration of MG7 antigen was detected by enzyme-linked immunosorbent assay (ELISA). This ELISA detection technology realizes the detection of the serum concentration of MG7 antigen through the "sandwich method": first, coat the 96-well plate with MG7 antigen monoclonal antibody (mouse) (1 μg / well); then add 10-fold diluted clinical Serum samples and serially diluted (2.5ng / mL–0.04ng / mL) human MG7 antigen solution (50 μl / well); then 50 μl of 0.47 μg / mL horseradish peroxidase (horseradish peroxidase, HRP) labeled mouse anti-human MG7 antigen polyclonal antibody (IgG). The immunoreaction plate containing the above mixture was incubated at room temperature under shaking conditions for 3 hours; after each reaction well was washed with washing solution, ...

Embodiment 3

[0139] Example 3 Joint detection of SEPT9 gene and RNF180 gene methylated DNA and MG7 antigen

[0140] From the data obtained in Example 1 and Example 2, analyze the methylation detection of SEPT9 gene and RNF180 gene and the detection data of MG7 antigen, the results show that: the detection sensitivity of SEPT9 gene and RNF180 gene methylation and MG7 antigen alone as gastric cancer markers However, if the SEPT9 gene, RNF180 gene methylation and MG7 antigen are combined to detect gastric cancer, the positive rate of detection will be greatly improved, and the sensitivity will reach 65.71% ( Figure 4 ). At the same time, the combined detection of SEPT9 gene, RNF180 gene methylation and MG7 antigen can maintain a good specificity for gastric cancer. The detection of 67 normal people showed that the specificity of the combined detection of the three was 88.06%. Therefore, the combined detection of SEPT9 gene, RNF180 gene and MG7 antigen has significant clinical application ...

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Abstract

The invention provides a composition for detecting gastric cancer, wherein the composition comprises: 1) a nucleic acid for detecting a methylation state in at least one region of a target gene, wherein the target gene is selected from an SEPT9 gene or fragments thereof and / or an RNF180 gene or fragments thereof; 2) an agent for detecting a concentration of an MG7 antigen in serum / plasma; the gastric cancer is detected by the methylation state of the target gene and a protein expression level of the target gene. The invention further provides a kit comprising the composition and a method for detecting the gastric cancer in vitro. Therefore, a method for analyzing DNA in a plasma sample by real-time PCR in the invention can conveniently detect the SEPT9 gene or the fragments thereof and / orthe RNF180 gene or the fragments thereof, and can rapidly and conveniently judge whether the sample is positive or not according to a cycle threshold (Ct) of the real-time PCR; further, the inventionprovides an accurate in-vitro detection method for detecting the concentration of the MG7 antigen in the serum / plasma of the sample to be detected.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a composition and its use in disease detection. Specifically, the invention relates to a composition for detecting gastric cancer, its corresponding kit and its use. Background technique [0002] Gastric cancer is a malignant tumor with high incidence in China. According to the data released by the National Cancer Prevention and Control Office: In 2015, there were 6.79x10 new cases of gastric cancer in my country. 5 Example, death 4.98x10 5 cases; both new cases and deaths rank second among common cancers. The death-to-case ratio of gastric cancer in my country is 0.733, which is very high. An important factor causing the high mortality rate and low cure rate of gastric cancer is the low diagnosis rate of early gastric cancer. Due to the lack of obvious and specific symptoms in early gastric cancer, most patients have entered the advanced stage of gastric cancer when they are diagn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886G01N33/574
CPCG01N33/57446G01N33/57488C12Q1/6886C12Q2600/154
Inventor 韩晓亮马竣周光朋徐海宋乐乐王建铭
Owner BIOCHAIN BEIJING SCI & TECH
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