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Autoantibody joint detection ELISA kit for gastric cardiac adenocarcinoma early screening

An autoantibody, combined detection technology, applied in the field of molecular biology and oncology, can solve problems such as no literature reports, achieve high sensitivity and specificity, reduce mortality, and reduce consumables.

Active Publication Date: 2019-08-30
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

P53 of the four TAAs has been reported to be related to the early diagnosis of cardiac adenocarcinoma, but Dock10, IVL, and CDH1 have not been reported in the literature in the diagnosis of cardiac adenocarcinoma
Application of Dock10, IVL, CDH1 and P53 four kinds of TAA to detect the expression of corresponding autoantibodies in the serum of patients with cardiac adenocarcinoma has not been reported

Method used

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  • Autoantibody joint detection ELISA kit for gastric cardiac adenocarcinoma early screening
  • Autoantibody joint detection ELISA kit for gastric cardiac adenocarcinoma early screening
  • Autoantibody joint detection ELISA kit for gastric cardiac adenocarcinoma early screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: the preparation of kit

[0028] 1. Experimental materials and reagents:

[0029] (1) 4 kinds of tumor-associated antigen proteins (Dock10, IVL, CDH1 and P53), purchased from Wuhan Aimeijie Technology Co., Ltd.;

[0030] (2) 96-well ELISA plate: 3590 (costar. USA);

[0031] (3) Coating solution: 50mM carbonate buffer solution, pH=9.6;

[0032] (4) Blocking solution: PBST buffer containing 2% (W / V) BSA;

[0033] (5) Sample diluent: PBST buffer containing 1% (W / V) BSA;

[0034] (6) Secondary antibody diluent: PBST buffer containing 1% (W / V) BSA;

[0035] (7) Enzyme-labeled secondary antibody: horseradish peroxidase-labeled RecA protein (Invitrogen);

[0036] (8) Washing solution: PBST (phosphate Tween) buffer containing 0.2% Tween 20;

[0037] (9) Positive control serum: P53 positive control serum, that is, the serum of patients with cardia adenocarcinoma who were positive for P53 antibodies by indirect ELISA and Western blot;

[0038] (10) Negative co...

Embodiment 2

[0063] Embodiment 2: the usage method of kit

[0064] 1. Serum sample incubation:

[0065] 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 wells of the 96-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.

[0066] 2. Enzyme-labeled secondary antibody incubation:

[0067] 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 96-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 well was discarded, and ...

Embodiment 3

[0072] Embodiment 3: Analysis of the diagnostic value of the kit of the present invention

[0073] Serum samples of patients with early cardiac adenocarcinoma and normal people were detected using the kit described in Example 1 of the present invention to evaluate and analyze the value of the kit of the present invention for screening and diagnosis of early cardiac adenocarcinoma.

[0074] 1. Sample source

[0075] A total of 160 serum samples were collected from the Henan Province Esophageal Cancer Key Open Laboratory of the First Affiliated Hospital of Zhengzhou University, including 80 normal human serum samples (control group) and 80 serum samples from early cardiac adenocarcinoma patients (early cardiac adenocarcinoma group). 80 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 80 normal subjects, there were 43 males and 37 females, with an averag...

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Abstract

The invention belongs to the technical field of the tumor medicine, and specifically discloses an autoantibody joint detection ELISA kit for gastric cardiac adenocarcinoma early screening. The kit comprises a solid-phase carrier and tumor-associated antigen coated on the solid-phase carrier; the tumor-associated antigen is composed of Dock10, IVL, CDH1 and P53; furthermore, the kit further comprises sample diluent, a second antibody, second antibody diluent, positive control serum, negative control serum, a developing solution, a stop solution and a washing solution. The ELISA kit disclosed bythe invention can effectively detect the gastric cardiac adenocarcinoma, especially the early gastric cardiac adenocarcinoma; the detection sensitivity is up to 87.5%, and the specificity reaches 75%; and the kit can be used for the large-scale screening of the asymptomatic population at the high incidence area of the gastric cardiac adenocarcinoma, thereby facilitating the gastric cardiac adenocarcinoma screening and early discovery of the asymptomatic high-risk group.

Description

technical field [0001] The invention relates to the fields of molecular biology and oncology, in particular to an autoantibody combined detection ELISA kit for early screening of cardia adenocarcinoma. Background technique [0002] Northern China, especially Linzhou in Henan Province and its adjacent Huixian, Anyang, Cixian and other places have the highest morbidity and mortality of cardia adenocarcinoma in the world. Due to the unclear pathogenesis of cardia adenocarcinoma, and the lack of early detection sensitive indicators and effective inspection methods for cardia adenocarcinoma, most patients have already developed the disease to the middle and late stages when they are diagnosed, and the prognosis is extremely poor. The 5-year survival rate of early cardia adenocarcinoma can reach more than 90%, while the 5-year survival rate of advanced patients is only about 10%. At present, cardiac adenocarcinoma is still one of the leading causes of tumor-related death in this ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57446
Inventor 王立东胡守佳赵学科宋昕韩月霞任景丽丁广成
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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