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Group of esophageal cancer detection markers and application thereof in preparation of esophageal cancer screening kit

A kit and technology for esophageal cancer, applied in the fields of molecular biology and oncology, can solve the problems of high false negative ratio, specificity and sensitivity to be improved, and achieve high yield, good photochemical stability and strong luminous intensity Effect

Active Publication Date: 2020-05-22
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PCR is not suitable for rapid detection. Although immunochromatography is fast, it has a large proportion of false negatives and can only be qualitative or semi-quantitative. Its specificity and sensitivity need to be improved

Method used

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  • Group of esophageal cancer detection markers and application thereof in preparation of esophageal cancer screening kit
  • Group of esophageal cancer detection markers and application thereof in preparation of esophageal cancer screening kit
  • Group of esophageal cancer detection markers and application thereof in preparation of esophageal cancer screening kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation of liquid-phase protein chip

[0033] The preparation process of the liquid phase protein chip of the present invention is as follows: figure 1 As shown, it mainly includes the following steps:

[0034] (1) Preparation of monoclonal antibody, including the following steps:

[0035] 1. Cell Fusion and Preparation of Hybridoma Cells

[0036] Female Balb / c mice aged 6-8 weeks were selected, and recombinant proteins of CA19-9, CYFRA21-1, CA125, FBXW7, and FAT1 were injected intraperitoneally as antigens, so that the antigens entered the peripheral immune organs through blood circulation or lymphatic circulation, stimulating the corresponding B lymphocytes activate, proliferate, and differentiate into sensitized B lymphocytes. After three days, adopt CO 2 The mice were killed by gas, and the spleen was removed under aseptic operation to prepare a spleen cell suspension. Take mouse myeloma cells and splenocytes in the logarithmic growth phase at a r...

Embodiment 2

[0053] Example 2 The method of using the protein chip

[0054] Take the liquid-phase protein chip stored in PBS buffer and shake it sufficiently to form a microparticle suspension. Add the serum sample to be tested and the protein chip in suspension state to the petri dish, mix well, and react in a 37°C incubator in the dark for 45 minutes; take out the petri dish, add an appropriate amount of microparticle suspension again, and mix well, React in a 37°C incubator protected from light for 30 minutes. The incubated samples were placed on a multifunctional flow cytometer analyzer for detection, and the relative expression levels of the five esophageal cancer markers in the serum samples to be tested were calculated.

Embodiment 3

[0055] Example 3 Analysis of the diagnostic value of the protein chip

[0056] The protein chip described in Example 1 of the present invention was used to detect the serum samples of patients with early esophageal cancer and normal people, so as to evaluate and analyze the value of the protein chip of the present invention for screening and diagnosis of early esophageal cancer.

[0057] 1. Sample source

[0058] Serum samples were collected from the Henan Province Esophageal Cancer Key Open Laboratory of the First Affiliated Hospital of Zhengzhou University, including 245 normal human serum samples as the control group and 245 early esophageal cancer patient serum samples as the early esophageal cancer group. Sera from 245 normal people came from the healthy check-up center of the laboratory's cooperative hospital, without any tumor-related evidence. Among the 245 normal subjects, there were 136 males and 109 females, with an average age of 57.6±8.5 years and an age range of...

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Abstract

The invention belongs to the field of molecular biology and oncology, and particularly relates to a group of esophageal cancer detection markers and application thereof in preparation of an esophagealcancer screening kit. The esophageal cancer detection markers serve a reagent capable of specifically detecting CA19-9, CYFRA21-1, CA125, FBXW7 and FAT1 or a group of gene expression products; the esophageal cancer early diagnosis kit is a liquid-phase protein chip kit. According to the esophageal cancer marker, a combination of the CA19-9 monoclonal antibody, the CYFRA21-1 monoclonal antibody, the CA125 monoclonal antibody, the FBXW7 monoclonal antibody and the FAT1 monoclonal antibody is used as the esophageal cancer marker for the first time, and is applied to preparation of a liquid-phaseprotein chip with high detection sensitivity and specificity for early esophageal cancer, the detection sensitivity of the liquid-phase protein chip to esophageal cancer is 93.5%, and the specificityof the liquid-phase protein chip to esophageal cancer is 73.9%.

Description

technical field [0001] The invention belongs to the fields of molecular biology and oncology, and in particular relates to a group of esophageal cancer detection markers and their application in the preparation of esophageal cancer screening kits. 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 esophageal cancer in the world. Extremely poor and still the leading cause of cancer-related death in the region. [0003] At present, the diagnosis of esophageal cancer is still based on gastrointestinal endoscopy. However, gastrointestinal endoscopy is invasive, inconvenient to operate, more painful for patients, and there are risks such as infection and bleeding, and the detection cost is relatively expensive. In recent years, scholars at home and abroad have begun to work on finding sensitive, specific, reliable and effective biomarkers for eso...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/577G01N33/574G01N33/543
CPCG01N33/54313G01N33/57407G01N33/57484G01N33/577G01N33/588
Inventor 王立东孟超龙宋昕赵学科范宗民王苒李贝韩雪娜靳艳杨苗苗
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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