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Application of dna carbon dot-silicon nano hydrogel material in the preparation of reagents for fluorescence detection of carcinoembryonic antigen in serum

A technology for fluorescence detection and carcinoembryonic antigen, which is applied in the application field of reagents, can solve the problems of poor stability, long construction time, and poor accuracy, and achieve good stability, short synthesis time, and reduce the cost of material construction

Active Publication Date: 2022-06-17
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these two methods have greatly improved the sensitivity of CEA detection, there are problems such as long construction time, poor stability, and poor accuracy in complex systems that need to be resolved urgently.

Method used

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  • Application of dna carbon dot-silicon nano hydrogel material in the preparation of reagents for fluorescence detection of carcinoembryonic antigen in serum
  • Application of dna carbon dot-silicon nano hydrogel material in the preparation of reagents for fluorescence detection of carcinoembryonic antigen in serum
  • Application of dna carbon dot-silicon nano hydrogel material in the preparation of reagents for fluorescence detection of carcinoembryonic antigen in serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Preparation of DNA carbon dots-silicon nanohydrogels

[0045] (1) Preparation of fluorescent carbon dots

[0046] S1. Weigh 0.6g of urea and 0.3g of citric acid in a beaker, add 10mL of deionized water to dissolve the powder completely, heat it in a microwave at 800W for 4min, and put it in a vacuum drying oven at 60℃ for 1h. Weigh the dry powder mass. Put the black powder into 40 mL of deionized water, centrifuge at 3000 rpm for 20 min, remove the filter residue, and take the supernatant, that is, the carbon dot stock solution.

[0047] (2) Preparation of SiNPs-CD-DNA

[0048] S2. Take 20 μL of the carbon dot stock solution prepared in step (1), dilute to 500 μL, and centrifuge at 12500 rpm for 20 min to prepare a carbon dot base solution for use.

[0049] S3. Add 20 μL 10 to the above carbon dot base solution -5 M 5' end-modified carboxyl group initiating DNA chain and 2 mg EDC, mixed with a mixer, and activated at room temperature for 15 min to obtain ...

Embodiment 2

[0067] Example 2: Detection of different concentrations of CEA by DNA carbon dots-silicon nanohydrogels

[0068] 1. Take 100 μL of the SiNPs-CD-MA-DNA carrier solution prepared in Example 1 into 18 centrifuge tubes, respectively numbered a to r.

[0069] 2. Add 20μL of 0ng / mL, 4x10 -6 ng / mL, 7x10 - 6 ng / mL, 1x10 -5 ng / mL, 4x10 -5 ng / mL, 7x10 -5 ng / mL, 1x10 -4 ng / mL, 4x10 -4 ng / mL, 7x10 -4 ng / mL, 1x10 -3 ng / mL, 4x10 -3 ng / mL, 7x10 -3 ng / mL, 1x10 -2 ng / mL, 4x10 -2 ng / mL, 7x10 -2 ng / mL, 0.1ng / mL, 0.4ng / mL, 1.0ng / mL analyte CEA standard solution.

[0070] 3. Put the above centrifuge tubes into which different concentrations of CEA standard solutions were added in sequence, and place them in a shaker at 37°C for 3 hours of shaking reaction.

[0071] 4. After the time is up, take out the above centrifuge tubes from the shaker, take 50 μL of the reaction solution from each centrifuge tube and place it in a fluorescence spectrophotometer cuvette. interval of the emissi...

Embodiment 3

[0074] Example 3: Fluorescence detection of carcinoembryonic antigen based on DNA carbon dots-silicon nanohydrogel materials Fluorescence signal contrast

[0075] Take 100 μL of the DNA carbon dots-silicon nanohydrogel prepared in Example 1 into 6 test tubes, one of which is a blank control, and add 20 μL of 4 test tubes to the concentration of 10 -6 ng / mL of bovine serum albumin (BSA), thrombin (thrombin), alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA), add 20 μL bovine serum albumin (BSA), thrombin to the last tube (thrombin), alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) concentrations were all 10 -6 ng / mL mixed solution, and put it into a shaker to react for 3 hours, and then measured the fluorescence intensity of 6 solutions with a molecular fluorometer. Repeat the experiment for many times and take the average value of the fluorescence intensity for subsequent data analysis.

[0076] The result is as Figure 5 As shown in the figure, the fluor...

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Abstract

The application provides a method for fluorescent detection of carcinoembryonic antigen in serum based on DNA carbon dot-silicon nano hydrogel material, which method includes the preparation of DNA carbon dot-silicon nano-hydrogel and DNA carbon dot-silicon nano-hydrogel Fluorescent detection of carcinoembryonic antigen, the preparation of DNA carbon dot-silicon nano hydrogel includes the preparation of fluorescent carbon dots, the preparation of SiNPs-CD-DNA, the combination of DNA and methacrylic acid, polymethacrylic acid (MAA)-DNA complex The synthesis of DNA carbon dot-silicon nano hydrogel. The DNA carbon dot-silicon nano hydrogel constructed by the present application requires simple reaction conditions, short construction time, and the use of it to detect CEA in serum has the advantages of wide detection range, high detection sensitivity, and good detection selectivity, and can be more Accurately detect the CEA content in serum samples, and provide help for early diagnosis of tumor patient lesions.

Description

technical field [0001] The invention relates to the application of DNA carbon dots-silicon nanometer hydrogel materials, in particular to the application of DNA carbon dots-silicon nanometer hydrogel materials in the preparation of reagents for fluorescence detection of carcinoembryonic antigens in serum. Background technique [0002] As a broad-spectrum tumor marker for clinical diagnosis of cancer, carcinoembryonic antigen (CEA) is closely related to colon cancer, rectal cancer, breast cancer, lung cancer and other malignant tumors. Determining its content in serum is of great significance for early diagnosis of malignant tumors and evaluation of curative effect. [0003] At present, enzyme-linked immunosorbent assay (ELISA) is commonly used in clinic to detect the content of CEA in serum. ELISA refers to immobilizing antigens or antibodies on solid carrier polyethylene microspheres, and qualitatively or quantitatively detecting target substances according to the enzyme-l...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/58G01N21/64
CPCG01N33/57473G01N33/582G01N33/587G01N21/6428G01N2021/6432
Inventor 纪小婷吕浩源丁彩凤
Owner QINGDAO UNIV OF SCI & TECH
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