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In-situ electrochemical immunosensor for simultaneously detecting four markers of breast cancer

An in-situ electrochemical and immunosensor technology, which is applied in the fields of biotechnology and medical treatment, can solve the problems of inaccurate detection results, harm to the human body, and long detection time, and achieve the effects of improved sensitivity, low detection limit, and high accuracy

Inactive Publication Date: 2021-06-29
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the deficiencies of the above-mentioned existing detection technologies, the purpose of the present invention is to provide an in-situ electrochemical immunosensor for simultaneous detection of four markers of breast cancer, which solves the problem that the existing detection methods need to extract living tissue, which is harmful to the human body, and the detection time Long time, high cost and inaccurate test results

Method used

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  • In-situ electrochemical immunosensor for simultaneously detecting four markers of breast cancer
  • In-situ electrochemical immunosensor for simultaneously detecting four markers of breast cancer
  • In-situ electrochemical immunosensor for simultaneously detecting four markers of breast cancer

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1 A method for preparing an in situ electrochemical immunosensor for simultaneous detection of four markers of breast cancer (1) Preparation of capture molecule silica-metal ion-antibody

[0039] 1) Take 28mL of distilled water, 65mL of ethanol and 7mL of ammonia water to form a mixed solution, then add 4mLTEOS and mix evenly, then magnetically stir at room temperature for 30min, and centrifuge and wash with absolute ethanol several times to obtain silica spheres.

[0040] 2) Add 0.022g of silica spheres to 2mL of ethanol solution, ultrasonicate for 30min to obtain a uniform silica sphere mixed solution, then add 0.4mL APTES dropwise to the above mixed solution, stir for 6h, mix and centrifuge to obtain The precipitate was washed 4 times with ethanol solution to obtain aminated silica spheres.

[0041] 3) Add 0.015 g of aminated silica spheres to 2 mL of Cd at a concentration of 10 mg / mL 2+ 、Cu 2+ , Pb 2+ and Hg 2+ and shaken at 30°C for 2 hours. After adsor...

Embodiment 2

[0050] Example 2. Method for molecular typing detection of breast cancer based on in situ electrochemical immunosensor for simultaneous detection of four markers of breast cancer

[0051] 1) Before use, use 1.0 μm and 0.05 μm alumina-water on a smooth polishing cloth to thoroughly polish the bare glassy carbon electrode (GCE) until a mirror surface is obtained, and then ultrasonicate the GCE in distilled water for 5 minutes to remove residual The alumina pellets were rinsed with distilled water and dried under nitrogen flow.

[0052]2) Incubate 10ul magnetic bead (MB) probes with proteins representing antigens of different breast cancer subtypes, shake gently at 37°C for 1 hour, magnetically separate and enrich the magnetic bead probes, and wash with PBS buffer to remove unfixed on the magnet Then disperse the immobilized magnetic bead probe in the mixed solution of 10ul silica-metal ion-antibody to carry out signal modification. After gently shaking and incubating for 0.5h, t...

Embodiment 3

[0057] Embodiment 3 condition optimization

[0058] The incubation times of magnetic bead probes and proteins representing antigens of different types of breast cancer were 20min, 30min, 40min, 50min, 60min, 70min, 80min, 90min and 100min, and other experimental steps were the same as in Example 2, and the experimental results were as follows: Figure 4 shown.

[0059] As can be seen from the figure, with the increase of the incubation time between the magnetic bead probe and the protein, HER2, ER, PR, Ki67 (Cd 2+ , Pb 2+ 、Cu 2+ , Hg 2+ ) The current signal also increased accordingly, and the current signal reached a plateau at 60min, 50min, 60min, 40min, and 70min respectively when the incubation time was 60min, 50min, 60min, 70min. In order to obtain sufficient detection current signals, the maximum incubation time of 70 min was selected as the optimal incubation time.

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Abstract

The invention discloses an in-situ electrochemical immunosensor for simultaneously detecting four markers of breast cancer. Molecular typing is performed on the breast cancer by detecting the four markers HER2, ER, PR and Ki67 in serum, and different metal ions are used as detection signals. And a magnetic bead-protein-silicon dioxide sandwich structure is formed for simultaneous detection of four markers. The sensor takes serum as a detection sample, breast cancer tissues do not need to be extracted, the operation is simple, the anti-interference capability is strong, a detection result is typed through an electrochemical signal of an electrochemical workstation, the repeatability is good, the accuracy is high, the detection limit is low, the whole reaction process can be shortened to 140 minutes, and the detection efficiency is greatly improved. The method has potential application value in diagnosis of different types of cancers.

Description

technical field [0001] The invention relates to the fields of biotechnology and medical technology, in particular to an in-situ electrochemical immunosensor for simultaneously detecting four markers of breast cancer. Background technique [0002] Breast cancer is one of the most common malignant tumors that endanger women's health. The incidence rate has increased significantly worldwide. According to statistics, the incidence rate accounts for 7-10% of various malignant tumors in the whole body, second only to uterine cancer in women. Cancer, its incidence is often related to heredity, and women between the ages of 40 and 60 have a higher incidence rate before and after menopause. It is one of the most common malignant tumors that seriously affects women's physical and mental health and even threatens their lives. Although the systemic treatment of breast cancer has made remarkable progress, many patients still suffer from tumor recurrence, metastasis and drug resistance, w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26
CPCG01N27/26
Inventor 侯长军张亚霍丹群徐发良李宁
Owner CHONGQING UNIV
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