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Colorectal cancer blood detection marker and application thereof

A technology for colorectal cancer and blood detection, which is applied in the field of tumor detection, can solve the problems of unsatisfactory blood detection results for colorectal cancer, and achieve the effect of improving sensitivity, high specificity, and maintaining specificity

Active Publication Date: 2022-03-01
ZHONGNAN HOSPITAL OF WUHAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the problem that the existing colorectal cancer blood detection effect is not ideal, and to provide a colorectal cancer blood detection marker and its application

Method used

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  • Colorectal cancer blood detection marker and application thereof
  • Colorectal cancer blood detection marker and application thereof
  • Colorectal cancer blood detection marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Extraction of DNA template:

[0030] When the sample used is a whole blood sample, the blood / cell / tissue genomic DNA extraction kit (catalogue number: DP304) of Tiangen Biochemical Technology (Beijing) Co., Ltd. is used. The volume of the used whole blood sample is 1 ml, and the extracted See the kit instructions for steps.

[0031] When the sample used is a plasma sample, use the nucleic acid extraction reagent of Wuhan Amison Life Science and Technology Co., Ltd. (Ehan Jibei No. 20210740) to extract plasma cfDNA. The volume of plasma used is 600 μL. For specific operations, refer to the kit instruction manual.

[0032] 2. Conversion of sulfite

[0033] The extracted whole blood genomic DNA or plasma cfDNA was subjected to bisulfite conversion, and the nucleic acid conversion kit used was the nucleic acid purification reagent of Wuhan Amison Life Science and Technology Co., Ltd.

[0034] 3. Methylation-specific PCR reaction

[0035] Perform methylation-specific P...

experiment example 1

[0049] 30 plasma samples from patients with advanced colorectal adenoma, 50 plasma samples from patients with colorectal cancer, and 30 plasma samples from healthy people from blood donation were collected from Zhongnan Hospital of Wuhan University. All patients were anonymized. According to the method described in Example 1, the plasma sample DNA was extracted, the extracted DNA was subjected to bisulfite conversion, and the converted DNA was used as a template to perform a methylation-specific PCR reaction, and the ITGA4 gene, ZNF829 gene and BEND5 gene were tested. The sensitivity of these three genes in adenoma and cancer samples and the specificity in healthy samples were calculated for the methylation levels in various plasma samples, and the results are shown in Table 3.

[0050] Table 3 Sensitivity and specificity data of three genes in adenoma, cancer and healthy plasma

[0051]

[0052] It can be seen from Table 3 that the sensitivity of methylation of the three g...

experiment example 2

[0054] 60 cases of whole blood samples from healthy people (different from the healthy people in Experimental Example 1) were collected from Zhongnan Hospital of Wuhan University, and all patients were anonymized. According to the method described in Example 1, the whole blood sample DNA was extracted, the extracted DNA was converted to bisulfite, and the converted DNA was used as a template to perform a methylation-specific PCR reaction, and the ITGA4 gene, ZNF829 gene and BEND5 were tested. For the methylation levels of the genes in 60 samples, the methylation positive rates of the three genes in the healthy samples were counted respectively, and the specificity was calculated. The results are shown in Table 4.

[0055] Table 4 The specificity of three genes in 60 cases of healthy people whole blood samples

[0056] gene name positive number specificity ITGA4 5 91.67％ ZNF829 12 80.00％ BEND5 6 90.00％

[0057] As shown in Table 4, the spe...

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Abstract

The invention discloses a colorectal cancer blood detection marker and application thereof, and belongs to the field of tumor detection. The colorectal cancer blood detection marker is a combination of BEND5 and TFPI2 genes, and the marker or a methylation detection reagent thereof can be used for preparing a colorectal cancer or precancerous lesion detection kit. When the marker is used for detecting colorectal cancer, high sensitivity and specificity are achieved, in a plasma sample, the sensitivity of BEND5 and TFPI2 methylation combination detection on I / II stage colorectal cancer is larger than 71%, the sensitivity of BEND5 and TFPI2 methylation combination detection on III / IV stage colorectal cancer is larger than 89%, and meanwhile the combination can effectively avoid false positive generated by gastric cancer, esophageal cancer and other digestive tract cancer species. The invention provides a new idea for noninvasive diagnosis of colorectal cancer.

Description

technical field [0001] The invention relates to the field of tumor detection, in particular to colorectal cancer blood detection markers and applications thereof. Background technique [0002] Colorectal cancer is one of the most common malignant tumors in my country and even in the world, mainly including colon cancer and rectal cancer. According to the national cancer statistics released by the National Cancer Center in 2019, the number of new cases of colorectal cancer in my country in 2015 was 376,300, and the number of deaths due to colorectal cancer was 191,000, ranking third in the incidence and mortality of malignant tumors. fifth place. The prognosis of colorectal cancer is related to the severity of the disease at the time of diagnosis. The 5-year survival rate of colorectal cancer patients in the United States is 67%, while it is only 31% in my country. From 2004 to 2013, the incidence of colorectal cancer in the United States decreased by 3% per year, and the ov...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6883C12N15/11
CPCC12Q1/6886C12Q1/6883C12Q2600/154
Inventor 林军张良禄卢长明
Owner ZHONGNAN HOSPITAL OF WUHAN UNIV
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