An ELISA kit for screening early cardia cancer

A kit and technology for cardiac cancer, applied in the fields of molecular biology and oncology, can solve the problems of rare applications, achieve high sensitivity and specificity, improve the detection rate, and achieve good technical reproducibility

Active Publication Date: 2020-02-11
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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  • An ELISA kit for screening early cardia cancer
  • An ELISA kit for screening early cardia cancer
  • An ELISA kit for screening early cardia cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: the preparation of kit

[0032] 1. Experimental materials and reagents:

[0033] (1) 6 kinds of tumor-associated antigen proteins (EYA4, ABL1, CD38, P53, NOTCH1, DLC1), purchased from Wuhan Amicate Technology Co., Ltd.;

[0034] (2) 48-well ELISA plate: purchased from Corning Company;

[0035] (3) Coating solution: 50mM carbonate buffer solution, pH=9.6;

[0036] (4) Blocking solution: PBST buffer containing 2% (W / V) BSA;

[0037] (5) Sample diluent: PBST buffer containing 1% (W / V) BSA;

[0038] (6) Secondary antibody diluent: PBST buffer containing 1% (W / V) BSA;

[0039] (7) Enzyme-labeled secondary antibody: horseradish peroxidase-labeled RecA protein (Invitrogen);

[0040] (8) Washing solution: PBST (phosphate Tween) buffer containing 0.2% Tween 20;

[0041] (9) Positive control serum: P53 positive control serum, that is, the serum of patients with cardia cancer whose P53 antibody is positive by indirect ELISA and Western blot;

[0042] (10) Nega...

Embodiment 2

[0067] Embodiment 2: the usage method of kit

[0068] 1. Serum sample incubation:

[0069] Dilute the serum sample to be tested with the sample diluent at a ratio of 1:500, and then add the diluted serum sample into 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 h, then discard the liquid in the reaction well, and wash with washing solution 3 times, each time for 3 min.

[0070] 2. Enzyme-labeled secondary antibody incubation:

[0071] 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 the liquid in the sample wells was discarded, an...

Embodiment 3

[0076] Embodiment 3: Analysis of the diagnostic value of the kit of the present invention

[0077] 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.

[0078] 1. Sample source

[0079] Serum samples were collected from the Henan Key Open Laboratory of Esophageal Cancer in the First Affiliated Hospital of Zhengzhou University, including 230 normal human serum samples (control group) and 230 serum samples from early cardiac cancer patients (early cardiac cancer group). 230 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 230 normal subjects, there were 123 males and 107 females, with an average age of 58.7±9.2 years and a range o...

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Abstract

The invention belongs to the technical field of tumor medicine, and particularly discloses an ELISA kit for screening of early cardia cancer. The kit comprises a solid phase carrier and a tumor-associated antigen coating the solid phase carrier, wherein the tumor-associated antigen is formed by EYA4, ABL1, CD38, P53, NOTCH1 and DLC1. Furthermore, the kit further comprises a sample diluent, a second antibody, a second antibody diluent, positive control serum, negative control serum, a developing solution, a stop solution and a washing solution. According to the ELISA kit disclosed by the invention, the cardia cancer, especially the early cardia cancer, can be effectively detected, the detection sensitivity is up to 92.2%, the specificity is up to 79.6%, the ELISA kit can be used for large-scale screening of an asymptomatic population in a high-incidence area of the cardia cancer, and is beneficial to screening and early discovery of the cardia cancer of the asymptomatic high-risk population.

Description

technical field [0001] The invention relates to the fields of molecular biology and oncology, in particular to an ELISA kit for early cardia cancer screening. Background technique [0002] With the development of society and the deterioration of living environment, cardiac cancer has become a type of malignant tumor that seriously threatens the health and safety of human life. 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, China is one of the areas with high incidence of esophageal cancer in my country. 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 act on the upper gastrointestinal mucosa for a long t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574
CPCG01N33/57446
Inventor 王立东王伟宋昕赵学科徐瑞华伍玥王启鸣
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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