A multi-gene combined detection reagent

A joint detection and multi-gene technology, applied in the field of biomedicine, can solve the problems of the sensitivity not reaching more than 90%, the overlap of colorectal cancer detection value and performance, and the difficulty of gene methylation markers, so as to achieve easy and reliable acquisition High diagnostic and detection sensitivity and specificity

Active Publication Date: 2021-12-21
CREATIVE BIOSCIENCES (GUANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing method for detecting a single methylation marker of colorectal cancer has the following two limitations: First, the existing research and product detection methods all detect one marker in one PCR reaction well, and the detection method is not able to Enables multiplex detection of methylated markers in one PCR reaction well
Second, when existing methylation markers for colorectal cancer detection such as SDC2, ITAG4, and Septin 9 detect colorectal cancer by detecting the methylation level of a single gene, the detection sensitivity is limited by genetic factors
The specificity of these individual markers for the detection of colorectal cancer can reach more than 90%, but the sensitivity when the specificity reaches more than 90% cannot reach more than 90%.
[0007] Although a single methylation marker of colorectal cancer can detect part of colorectal cancer, it is necessary to combine colorectal cancer-related markers to detect as many colorectal cancer patients as possible through molecular biology methods. The problem is: 1. The detection value of most colorectal cancer gene methylation markers for colorectal cancer is overlapping, that is, the detection status of the same colorectal cancer patient sample is consistent among most methylation markers of
Therefore, it is very difficult to screen out gene methylation markers that can complement each other in the detection of colorectal cancer, requiring a lot of research work
In addition, the more gene methylation markers, the false positive phenomenon of the detection system will also be superimposed, and the specificity of detection will decrease

Method used

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  • A multi-gene combined detection reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] According to Table 1, three pairs of primers for SDC2 were designed to study the specificity of amplification.

[0138] Table 1 Primers for SDC2

[0139]

[0140]

[0141] The above three pairs of primers were used to detect the DNA of stool samples from 2 cases of colorectal cancer patients and 2 cases of normal human stool samples, and analyze the specificity of amplification.

[0142] The amplification curve of 3 pairs of primers amplifying SDC2 is as follows Figure 1-3 shown.

[0143] The results showed that SDC2 primers had a better amplification specificity for S0; while the other two pairs of primers had little difference between negative and positive samples in their amplification curves, and could not effectively distinguish between negative and positive samples.

Embodiment 2

[0145] According to Table 2, three pairs of COL4A1 / COL4A2 primers were designed to study the specificity of amplification.

[0146] Table 2 Primers for COL4A1 / COL4A2

[0147]

[0148] The above 3 pairs of primers were used to detect the DNA of stool samples from 3 cases of colorectal cancer patients and 3 cases of normal human stool samples, and analyze the specificity of amplification.

[0149] The amplification curves of 3 pairs of primers amplifying COL4A1 / COL4A2 are as follows: Figure 4-6 shown.

[0150] The results showed that COL4A1 / COL4A2 primer pair C0 and C1 had better amplification specificity; while primer pair C2 had no amplification for COL4A1 / COL4A2.

Embodiment 3

[0152] According to Table 3, three pairs of primers for ITGA4 were designed to study the specificity of amplification.

[0153] Table 3 Primers of ITGA4

[0154]

[0155]

[0156] The above 3 pairs of primers were used to detect the DNA of stool samples from 3 cases of colorectal cancer patients and 3 cases of normal human stool samples, and analyze the specificity of amplification.

[0157] The amplification curve of 3 pairs of primers to amplify ITGA4 is as follows Figure 7-9 shown.

[0158] The results showed that the primer pair I0 and I2 of ITGA4 had better amplification specificity; while the amplification curve of primer pair I1 had little difference between negative and positive samples, and could not effectively distinguish between negative and positive samples.

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Abstract

The invention belongs to the field of biomedicine, in particular to tumor markers, methylation detection reagents, kits and applications thereof. Through research, the present invention finds that colorectal cancer specimens can be well distinguished from feces specimens by detecting the methylation levels of the combined promoter regions of SDC2, COL4A1 / COL4A2 and ITGA4 genes. The present invention utilizes the methylation reagent containing the gene combination to detect colorectal cancer, and the detection sensitivity and specificity to colon cancer are very high, both of which can reach more than 90%.

Description

technical field [0001] The invention belongs to the field of biomedicine, and particularly relates to primers, capture reagents, nucleic acid probes, methylation detection reagents, kits and applications thereof. [0002] Background of the invention [0003] Colorectal cancer, also known as colorectal cancer, is a common malignant tumor of the digestive tract. Its incidence rate is increasing year by year in my country. In some coastal areas of my country such as Shanghai and Guangzhou, the incidence rate of colorectal cancer has jumped to the second place, second only to lung cancer. It is currently believed that the formation of colorectal cancer is the result of the accumulation of genetic and epigenetic defects. The onset of colorectal cancer is hidden in the early stage, and there are often no obvious symptoms. Symptoms such as blood in the stool, abdominal pain, and diarrhea may appear in the late stage. And when symptoms appear, it is often late, which brings great m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6886
CPCC12Q1/6886C12Q2600/118C12Q2600/154
Inventor 吴孝林刘相林罗茵邹鸿志
Owner CREATIVE BIOSCIENCES (GUANGZHOU) CO LTD
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