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An ELISA kit for early diagnosis of esophageal squamous cell carcinoma

A technology of esophageal squamous cell carcinoma and a kit, which is applied in the field of medicine and biology, can solve the problems of high cost, limited popularization and application of endoscopic screening, low efficiency, etc., and achieve the effect of improving the detection rate and the correct rate.

Active Publication Date: 2020-10-09
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the trauma, high cost and low efficiency of endoscopic screening (for example, endoscopic screening in routine high-incidence areas is as above "high-risk groups", the detection rate of early cancer is only about 2%, and about 90% of asymptomatic "high-risk groups" "crowd" are "companion inspection"), which limits the promotion and application of endoscopic screening in the early detection of esophageal squamous cell carcinoma in asymptomatic high-risk groups

Method used

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  • An ELISA kit for early diagnosis of esophageal squamous cell carcinoma
  • An ELISA kit for early diagnosis of esophageal squamous cell carcinoma
  • An ELISA kit for early diagnosis of esophageal squamous cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: the preparation of kit

[0037] According to the principle of indirect ELISA, the invention prepares a multi-link ELISA kit for rapid detection of esophageal squamous cell carcinoma. The principle of indirect enzyme-linked immunoassay is to connect the antigen to a solid-phase carrier, and the antibody to be tested in the sample combines with it to form a solid-phase antigen-test antibody complex, and then use the enzyme-labeled secondary antibody and the solid-phase antigen-test antibody to The antibody in the complex is combined to form a solid-phase antigen-test antibody-enzyme-labeled secondary antibody complex, and then measure the degree of color development after adding the substrate to determine the content of the antibody to be tested (see figure 2 ).

[0038] 1. Experimental materials and reagents:

[0039] (1) 6 tumor-associated antigen proteins (HMGB1, TLR4, TAK1, ATG5, SLC22A3, CD38), purchased from Wuhan Amicate Technology Co., Ltd.;

[0...

Embodiment 2

[0073] Embodiment 2: the usage method of kit

[0074] 1. Serum sample incubation:

[0075] Dilute the serum sample to be tested with the sample diluent at a ratio of 1:100, and then add the diluted serum sample into the reaction wells 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.

[0076] 2. Enzyme-labeled secondary antibody incubation:

[0077] Dilute the horseradish peroxidase-labeled RecA protein with the secondary antibody diluent at a ratio of 1:80000, 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 well was discarded, an...

Embodiment 3

[0082] Embodiment 3: Analysis of the diagnostic value of the kit of the present invention

[0083] Serum samples from patients with early esophageal squamous cell carcinoma and normal people were detected with the kit described in Example 1 of the present invention, so as to evaluate and analyze the value of the kit of the present invention for screening and diagnosis of early esophageal squamous cell carcinoma.

[0084] 1. Sample source

[0085] Serum samples were collected from the Henan Key Open Laboratory of Esophageal Squamous Cell Carcinoma in the First Affiliated Hospital of Zhengzhou University, including 150 normal human serum samples (control group) and 150 serum samples from patients with early ESCC (early ESCC group). Among the 150 normal subjects, there were 79 males and 71 females, with an average age of 55.8±9.1 years and a range of 39-80 years. Among the 150 patients with esophageal squamous cell carcinoma, there were 82 males and 68 females, with an average a...

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Abstract

The invention belongs to the technical field of medical biology, and particularly discloses an ELISA kit for early diagnosis of esophageal squamous cell carcinoma. The kit comprises a solid phase carrier and a tumor-associated antigen coated on the solid phase carrier, and the tumor-associated antigen is composed of HMGB1, TLR4, TAK1, ATG5, SLC22A3 and CD38. Furthermore, the kit also 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. The ELISA kit provided by the invention can effectively detect esophageal squamous cell carcinoma, especially early esophageal squamous cell carcinoma, has detection sensitivity as high as 92.7% and specificity as high as 84.0%,can be used for large-scale screening and diagnosis of asymptomatic people in esophageal squamous cell carcinoma high-incidence areas, and is beneficial to screening and early discovery of asymptomatic high-risk people.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and in particular relates to an ELISA kit for early diagnosis of esophageal squamous cell carcinoma. Background technique [0002] Esophageal squamous cell carcinoma (ESCC) is a malignant tumor that occurs in human esophageal epithelial tissue. It reduces people's quality of life, seriously endangers people's life and health, and brings heavy disasters to patients' families and the country. come a heavy burden. ESCC is a common malignant tumor of the digestive system worldwide, and there are typical high-incidence areas of esophageal squamous cell carcinoma in all countries in the world. The high-incidence regions of esophageal squamous cell carcinoma in China have a prominent clustering characteristic. Since the 1970s, many typical high-incidence regions of esophageal squamous cell carcinoma have been discovered, such as Linxian County in Henan Province, Cixian County in Hebei Prov...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/543
CPCG01N33/543G01N33/57407
Inventor 王立东吉佳佳赵学科宋昕雷玲玲韩雪娜范宗民王苒李贝王盼盼
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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