Reagent, kit and device for detecting tumor micro-environment of colorectal cancer as well as application of reagent

A tumor microenvironment and detection reagent technology, which is applied in the field of colorectal cancer tumor microenvironment detection reagents

Pending Publication Date: 2020-06-05
ZHENYUE BIOTECHNOLOGY JIANGSU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The main purpose of the present invention is to provide a colorectal cancer tumor microenvironment detection reagent, kit, device and appl

Method used

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  • Reagent, kit and device for detecting tumor micro-environment of colorectal cancer as well as application of reagent
  • Reagent, kit and device for detecting tumor micro-environment of colorectal cancer as well as application of reagent
  • Reagent, kit and device for detecting tumor micro-environment of colorectal cancer as well as application of reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Four cases of stage II patients were selected: 2 cases of rectal adenocarcinoma, 1 case of colonic mucinous adenocarcinoma, and 1 case of colonic adenocarcinoma with mucinous adenocarcinoma.

[0097] Table 4:

[0098] Numbering tumor site Clinical stage T stage Follow-up time (year) whether to die 1 rectum II 4 6.7 no 2 high rectum II 3 4.0 no 3 Sigmoid colon II 4 1.7 no 4 ascending colon II 4 1.2 yes

[0099] Multiplex immunohistochemical analysis of immune cells and immune checkpoint molecular detection was performed on the paraffin sections of the tumor tissues of the above four cases. 2 slices per patient, a total of 2 staining panels, and a total of 10 molecules were labeled. The staining order of each molecule in 10 staining molecules and 2 panels is: Panel1: CD45RO, PD-L1, CD8, CD3, PD1; Panel2: FoxP3, CD163, CD68, CD4, PDL1, CD57.

[0100] Perform multiple immunohistochemical analysis of i...

Embodiment 2

[0133] DNA samples from 120 stage II CRC cases were used to construct a genome-wide sequencing library, and high-throughput sequencing was performed to detect whether there was a mutation in the POLE gene. The test results were as follows: 20 cases of POLE gene mutation samples among 120 samples, and 20 SNP site mutations were involved in total, and 100 cases of POLE gene wild type samples.

[0134] The POLE gene wild-type samples and POLE gene mutant samples were divided into two groups, and the expression of each molecular marker was detected by the same multiple immunohistochemical method as in Example 1, and the corresponding immune responses of the POLE gene mutant and wild-type samples were detected. The positive rate expression of microenvironmental marker cells. See the test results figure 1 , figure 1 The percentage of positive cells for the different genotypes of POLE and expression of each molecular marker or bimolecular marker is shown.

Embodiment 3

[0136] The same multiple immunohistochemical detection method as in Example 1 was used to detect the positive rate of cells of each molecular marker in the tumor epithelium (CT) in 120 cases of stage II CRC. The test results are shown in Table 2 and Table 3.

[0137] From the above description, it can be seen that the above-mentioned embodiments of the present invention have achieved the following technical effects: the use of the detection reagent of the POLE gene provided by the application and the molecular markers in any of the above-mentioned combinations and the combination thereof are effective against colorectal cancer. When testing, the tumor microenvironment of the corresponding patient can be detected more accurately, so that the survival and prognosis of the sample to be tested can be guided and predicted more effectively.

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Abstract

The invention provides a reagent, a kit and a device for detecting tumor micro-environment of colorectal cancer as well as application of the reagent. The detection reagent comprises a POLE gene mutation detection reagent and an antibody of at least one molecular marker of at least one of the following combinations such as a combination 1: CD3, CD8, CD45RO, PD-1 and PD-L1 and a combination 2: CD4,FOXP3, CD68, CD163, PD-L1 and CD57. By utilizing the reagent, the kit and the device for detecting tumor micro-environment of colorectal cancer as well as the application of the reagent disclosed bythe invention, findings prove that POLE gene mutation is obviously related with high expression of a part of the molecular markers, and a relation between the POLE gene mutation and the tumor micro-environment of the colorectal cancer is determined, so that a multi-immunohistochemical method is conveniently and comprehensively utilized, and simultaneously several molecular markers are marked, andthe advantages that the sensitivity of detection on the tumor micro-environment of a pancreatic patient is high and the specificity is high are also utilized, so that whether a detected object has 5-year survival rate of higher probability can be relatively and accurately predicted.

Description

technical field [0001] The invention relates to the field of multiple immunohistochemical detection, in particular to a detection reagent, kit, device and application of colorectal cancer tumor microenvironment. Background technique [0002] The existing immunohistochemistry commonly used diaminobenzidine (ie 3,3,-diaminobenzidine, DAB) color development, the principle is because diaminobenzidine is the chromogenic substrate of peroxidase, in the presence of hydrogen peroxide Under the condition of electron loss, the color change accumulates, forming a tan insoluble product. DAB is mainly combined with the NH2 or SH group of the protein to form a stable N-N bond or N-S bond, and then the chromogenic group in DAB can display the color and mark it on the exposed protein, thereby showing the protein distribution of the target cell and types etc. The prior art generally marks one molecule on one slice. [0003] Multiple immunohistochemistry (mIHC, multiple immnohistochemistry...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/574
CPCC12Q1/6886G01N33/57419G01N33/57484C12Q2600/156
Inventor 张恒辉李亚丹赵海涛林健振钱娟娟
Owner ZHENYUE BIOTECHNOLOGY JIANGSU CO LTD
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