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An autoantibody combined detection ELISA kit for early screening of esophageal squamous cell carcinoma

A technology of combined detection of autoantibodies, which is applied in the fields of molecular biology and oncology, can solve problems that have not been reported before, and achieve the effects of convenient use, reduced mortality, and less consumables

Active Publication Date: 2020-02-21
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 esophageal squamous cell carcinoma, but TP53, CDKN2B and NPM1 have not been reported in the literature in the diagnosis of esophageal squamous cell carcinoma
The application of four TAAs, P53, TP53, CDKN2B and NPM1, to detect the expression of corresponding autoantibodies in the serum of patients with esophageal squamous cell carcinoma has not been reported.

Method used

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  • An autoantibody combined detection ELISA kit for early screening of esophageal squamous cell carcinoma
  • An autoantibody combined detection ELISA kit for early screening of esophageal squamous cell carcinoma
  • An autoantibody combined detection ELISA kit for early screening of esophageal squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: the preparation of kit

[0031] 1. Experimental materials and reagents:

[0032] (1) Four kinds of tumor-associated antigen proteins (P53, TP53, CDKN2B and NPM1), purchased from Wuhan Aimeijie Technology Co., Ltd.;

[0033] (2) 96-well ELISA plate: 3590 (costar. USA);

[0034] (3) Coating solution: 50mM carbonate buffer solution, pH=9.6;

[0035] (4) Blocking solution: PBST buffer containing 2% (W / V) BSA;

[0036] (5) Sample diluent: PBST buffer containing 1% (W / V) BSA;

[0037] (6) Secondary antibody diluent: PBST buffer containing 1% (W / V) BSA;

[0038] (7) Enzyme-labeled secondary antibody: horseradish peroxidase-labeled RecA protein (Invitrogen);

[0039] (8) Washing solution: PBST (phosphate Tween) buffer containing 0.2% Tween 20;

[0040] (9) Positive control serum: P53 positive control serum, that is, the serum of patients with esophageal squamous cell carcinoma who were positive for P53 antibodies by indirect ELISA and Western blot;

[0041]...

Embodiment 2

[0066] Embodiment 2: the usage method of kit

[0067] 1. Serum sample incubation:

[0068] 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.

[0069] 2. Enzyme-labeled secondary antibody incubation:

[0070] 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, and incubated in a constant temperature incubator at 37°C for 50 minutes, then the liquid in the reaction well was discarded, and wa...

Embodiment 3

[0075] Embodiment 3: Analysis of the diagnostic value of the kit of the present invention

[0076] Serum samples from patients with early esophageal squamous cell carcinoma 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 esophageal squamous cell carcinoma.

[0077] 1. Sample source

[0078] A total of 160 serum samples were collected from the Henan Key Open Laboratory of Esophageal Cancer, the First Affiliated Hospital of Zhengzhou University, including 80 normal human serum samples (control group) and 80 serum samples from early ESCC patients (early ESCC group). The sera of 80 normal people came from the healthy check-up population of the physical examination center of the laboratory's cooperative hospital, without any evidence of tumor-related. 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 medical oncology, and particularly discloses an autoantibody joint detection ELISA kit for early screening of esophageal squamous carcinoma. The kit comprises a solid-phase carrier and a tumor-associated antigen coated on the solid-phase carrier, wherein the tumor-associated antigen consists of P53, TP53, CDKN2B and NPM1. Furthermore, the kit comprises sample diluent, a second antibody, second antibody diluent, positive control serum, negative control serum, developing solution, stop solution and washing solution. The ELISA kit is capable of effectively detecting the esophageal squamous carcinoma, especially the early esophageal squamous carcinoma, has detection sensitivity up to 88.8% and specificity up to 86.3%, can be used for the large-scale screening of asymptomatic groups in high-risk areas of the esophageal squamous carcinoma, and is beneficial for the esophageal squamous carcinoma screening and early discovery of the asymptomatichigh-risk groups.

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 esophageal squamous cell carcinoma. Background technique [0002] Esophageal cancer is one of the most common malignant tumors in the world, and its mortality rate ranks sixth. More than half of the approximately 500,000 new cases of esophageal squamous cell carcinoma in the world occur in China, which is 100 times higher than that in Western countries. China is the country with the highest morbidity and mortality of esophageal squamous cell carcinoma. According to statistics, there are about 250,000 newly diagnosed cases in my country every year, accounting for half of the total number of new cases of esophageal squamous cell carcinoma in the world. In my country, the incidence of esophageal squamous cell carcinoma is not only high, but also has significant regional differences. For example, northe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574
CPCG01N33/57407
Inventor 王立东程锟宋昕赵学科冯常炜朱文亮周建炜
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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