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A multiplex immunohistochemical analysis kit for liver cancer and its use and application

A technology of immunohistochemistry and analytical reagents, which is applied in the direction of analytical materials, biological testing, instruments, etc., can solve problems such as influence and inaccurate detection results, and achieve the effects of high detection efficiency, elimination of interference, and high accuracy

Active Publication Date: 2020-05-29
ZHENYUE BIOTECHNOLOGY JIANGSU CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are currently some immunohistochemical staining methods for multiple markers, these multiple recombination staining methods will affect the staining formed in the previous round during elution, often need to add staining enhancers, etc., and the detection results are not accurate enough

Method used

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  • A multiplex immunohistochemical analysis kit for liver cancer and its use and application
  • A multiplex immunohistochemical analysis kit for liver cancer and its use and application
  • A multiplex immunohistochemical analysis kit for liver cancer and its use and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] A total of 20 liver cancer patients with poor tumor tissue immunotherapy (shorter overall survival (OS) (number 1-10)) and better immunotherapy (longer OS (number 11-20)) were selected. 10 cases, the specific survival time is shown in Table 1.

[0106] Table 1 Survival period of 20 patients with liver cancer

[0107]

[0108] Multiplex immunohistochemical analysis of proteins in the tumor microenvironment was performed as follows. Each patient had 2 slices (panel 1 and panel 2), and each slice was labeled with 5 molecules. The staining sequence is: Panel1: CD66B, PanCK, PDL1, CD163, CD8; Panel 2: PD-1, PanCK, PDL1, CD8, CD68. The primary antibody, secondary antibody, and fluorescent dye information during the experiment are shown in Table 2:

[0109] Table 2 Primary antibody, secondary antibody and fluorescent dye information

[0110]

[0111] Example operation steps:

[0112] 1. Bake 20 paraffin sections in a 60°C incubator for 120 minutes;

[0113] 2. Dewa...

Embodiment 2

[0158] A multiple immunohistochemical analysis kit for liver cancer, including a monoclonal antibody group, antigen retrieval solution, horseradish peroxidase-labeled secondary antibody, hydrogen peroxide, fluorophore-labeled tyrosine and nuclear staining agent;

[0159] The monoclonal antibody group is: PanCK monoclonal antibody, PDL1 monoclonal antibody, CD8 monoclonal antibody, CD66B monoclonal antibody and CD163 monoclonal antibody;

[0160] The fluorescent groups are 520-FITC, 570-Cy3, 620-Cy3.5, 650-Cy5 and 690-Cy5.5;

[0161] The kit also includes an antiquencher, a washing agent and a blocking solution.

Embodiment 3

[0163] A multiple immunohistochemical analysis kit for liver cancer, including a monoclonal antibody group, antigen retrieval solution, horseradish peroxidase-labeled secondary antibody, hydrogen peroxide, fluorophore-labeled tyrosine and nuclear staining agent;

[0164] The monoclonal antibody group is: PanCK monoclonal antibody, PDL1 monoclonal antibody, CD8 monoclonal antibody, PD1 monoclonal antibody and CD68 monoclonal antibody;

[0165] The fluorescent groups are 520-FITC, 570-Cy3, 620-Cy3.5, 650-Cy5 and 690-Cy5.5;

[0166] The kit also includes an antiquencher, a washing agent and a blocking solution.

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Abstract

The invention provides a multiple immunohistochemical analysis kit for liver cancer, and a using method and application thereof, and relates to the technical field of multiple immunohistochemistry. The multiple immunohistochemical analysis kit limits that monoclonal antibody groups comprise a first monoclonal antibody group and a second monoclonal antibody group; the first monoclonal antibody group comprises a PanCK monoclonal antibody, a PDL1 monoclonal antibody, a CD8 monoclonal antibody, a CD66B monoclonal antibody and a CD163 monoclonal antibody; the second monoclonal antibody group comprises a PanCK monoclonal antibody, a PDL1 monoclonal antibody, a CD8 monoclonal antibody, a PD1 monoclonal antibody and a CD68 monoclonal antibody. According to the multiple immunohistochemical analysiskit provided by the invention, the paraffin sections of the human liver tissue in vitro are used as samples for detecting, and thus the effectiveness of the immune checkpoint inhibitor acting on theliver cancer is predicted.

Description

technical field [0001] The invention relates to the technical field of multiple immunohistochemistry, in particular to a multiple immunohistochemical analysis kit for liver cancer and its use method and application. Background technique [0002] Liver cancer is the second leading cause of cancer-related death worldwide. It is estimated that in 2012, there were 782,000 new cases of liver cancer worldwide, and 745,000 deaths from liver cancer, of which about 50% occurred in China. Hepatocellular carcinoma (HCC), which accounts for 90% of primary liver cancers, can be caused by chronic infection with hepatitis B virus (HBV) or hepatitis C virus (HCV), alcohol abuse, and metabolic syndrome associated with diabetes and obesity. Over the past decade, significant advances have been made in the molecular pathogenesis of HCC, but current therapeutic options are still very limited. Only a small number of patients with liver cancer are diagnosed at an early stage, when treatment meth...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/577G01N33/58
Inventor 张恒辉陈衍辉罗红丽王雅婷胡莹刘阳石伟陆吉麟
Owner ZHENYUE BIOTECHNOLOGY JIANGSU CO LTD
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