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Application of a group of tumor-associated antigens in the preparation of early screening kits for cardia cancer

A tumor-associated antigen, cardia cancer technology, applied in the fields of molecular biology and oncology, can solve the problems of rare application, achieve the effects of high sensitivity and specificity, less consumables, and convenient use

Active Publication Date: 2020-09-01
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of multiple autoantibody combined detection methods and corresponding kits for the early diagnosis of cardia cancer is relatively rare.

Method used

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  • Application of a group of tumor-associated antigens in the preparation of early screening kits for cardia cancer
  • Application of a group of tumor-associated antigens in the preparation of early screening kits for cardia cancer
  • Application of a group of tumor-associated antigens in the preparation of early screening kits for cardia cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The preparation of embodiment 1 kit

[0030] 1. Experimental materials and reagents:

[0031] (1) Six tumor-associated antigen proteins (CD38, CCND1, RB1, KMT2D, FAT1, EP300), purchased from Wuhan Amicate Technology Co., Ltd.;

[0032] (2) 48-well ELISA plate: purchased from Corning;

[0033] (3) Coating solution: 50mM carbonate buffer, pH=9.6;

[0034] (4) Blocking solution: PBST buffer containing 2% (W / V) BSA;

[0035] (5) Sample diluent: PBST buffer containing 1% (W / V) BSA;

[0036] (6) Secondary antibody diluent: PBST buffer containing 1% (W / V) BSA;

[0037] (7) Enzyme-labeled secondary antibody: horseradish peroxidase-labeled RecA protein (Invitrogen);

[0038] (8) Washing solution: PBST buffer solution with pH=7.4 and containing 0.05v% Tween-20.

[0039] (9) Positive control serum: KMT2D positive control serum, that is, the serum of patients with gastric cardia cancer whose KMT2D antibody is positive by indirect ELISA and Western blot;

[0040](10) Negative...

Embodiment 2

[0065] Embodiment 2: the usage method of kit

[0066] 1. Serum sample incubation:

[0067] Dilute the serum sample to be tested with the sample diluent at a ratio of 1:500, and then add the diluted serum sample to the reaction well of the 48-well microplate plate coated with antigen, with a sample volume of 100 μL / well, Place in a constant temperature incubator at 37°C and incubate for 1 hour, then discard the liquid in the reaction well, and wash with washing solution 3 times, 3 minutes each time.

[0068] 2. Enzyme-labeled secondary antibody incubation:

[0069] Dilute the horseradish peroxidase-labeled RecA protein with the secondary antibody diluent at a ratio of 1:40000, and then add the diluted horseradish peroxidase-labeled RecA protein to the reaction wells of the 48-well microtiter plate In this method, the sample volume was 100 μL / well, placed in a 37°C constant temperature incubator and incubated for 50 minutes, then discarded the liquid in the sample well, and wa...

Embodiment 3

[0074] Embodiment 3: Analysis of the diagnostic value of the kit of the present invention

[0075] The kit described in Example 1 of the present invention was used to detect the serum samples of patients with early cardiac cancer and normal people, so as to evaluate and analyze the value of the kit of the present invention for screening and diagnosis of early cardiac cancer.

[0076] 1. Sample source

[0077] Serum samples were collected from the Henan Province Key Open Laboratory of Esophageal Cancer, the First Affiliated Hospital of Zhengzhou University, including 190 normal human serum samples (control group) and 190 serum samples from early cardiac cancer patients (early cardiac cancer group). 190 cases of normal human serum were collected from the healthy physical examination center of the laboratory's cooperative hospital, without any tumor-related evidence. Among the 190 normal subjects, there were 108 males and 92 females, with an average age of 58.7±9.2 years and a r...

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Abstract

The invention belongs to the field of molecular biology and oncology, and particularly relates to an application of a group of tumor-associated antigens in preparation of a cardia cancer early screening kit. The invention discloses a combination of tumor-associated antigens CD38, CCND1, RB1, KMT2D, FAT1 and EP300 for preparing a cardia cancer early screening kit. The ELISA kit for screening the early cardia cancer comprises a solid-phase carrier and tumor-associated antigens coating the solid-phase carrier, and the tumor-associated antigens are CD38, CCND1, RB1, KMT2D, FAT1 and EP300. According to the invention, the six tumor-associated antigens including CD38, CCND1, RB1, KMT2D, FAT1 and EP300 are taken as a combination for the first time, the combination is used for detecting the cardiacancer, especially the early cardia cancer, and the kit has the detection sensitivity of 94.74% and the specificity of 84.21%.

Description

technical field [0001] The invention belongs to the fields of molecular biology and oncology, and specifically relates to the application of a group of tumor-associated antigens in the preparation of an early screening kit for cardiac cancer. Background technique [0002] With the development of society and the change of environment, cardia cancer has become a kind of malignant tumor that seriously threatens human life, health and safety. Cardiac cancer refers to the cancer that occurs in the anatomical part of the cardia, which is generally considered to be located within 2 cm below the esophagogastric junction in China. Linzhou City, Henan Province is one of the areas with a high incidence of esophageal cancer in China. Studies have found that there is also a high incidence of cardia cancer in areas with a high incidence of esophageal cancer. This phenomenon suggests that there may be strong carcinogenic factors in the local living environment. These carcinogenic factors ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/558G01N33/535
CPCG01N33/535G01N33/558G01N33/57407
Inventor 王立东杨媛啧宋昕赵学科袁翎杨道科李健徐瑞华韩文莉雷玲玲
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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