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A fluorescent detection device assembled with induced DNA and a method for detecting the content of small molecules using it

A technology of small molecules and DNA sequences, which is applied in the field of fluorescent detection devices based on binding-induced DNA assembly and the detection of small molecule content, can solve the problem of high fluorescence background value, and achieve the effects of simple operation, ingenious and reasonable structure, and easy synthesis

Active Publication Date: 2022-04-15
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method has the disadvantage of high fluorescence background value

Method used

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  • A fluorescent detection device assembled with induced DNA and a method for detecting the content of small molecules using it
  • A fluorescent detection device assembled with induced DNA and a method for detecting the content of small molecules using it
  • A fluorescent detection device assembled with induced DNA and a method for detecting the content of small molecules using it

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0052] Preparation of recognition elements modified by DNA and antibodies; preparation of signal elements for fluorescently labeled antigens to obtain detection devices;

[0053] And the detection device obtained above is used for the detection of the small molecule content of the sample.

Embodiment

[0056] Detection device and detection method based on binding-induced DNA assembly fluorescence detection of aflatoxin B1:

[0057] Prepare the identification element:

[0058] Colloidal gold modified with anti-aflatoxin B1 monoclonal antibody and auxiliary DNA, particle size 20nm;

[0059] The sequence of the helper DNA is:

[0060] 5'-Mercapto-TTTTT AGCAGTG T-3';

[0061] The colloidal gold was first adjusted to pH 8.8 with potassium carbonate solution, then the monoclonal antibody against aflatoxin B1 was added, and reacted at room temperature for 30 minutes. The molar ratio of monoclonal antibody and colloidal gold is 110:1. Add helper DNA and incubate at room temperature for 10 min. The molar ratio of helper DNA to colloidal gold is 500:1. Add polyethylene glycol 20000 with a final concentration of 2%, and incubate at room temperature for 30 minutes; add sodium chloride with a final concentration of 500 mM, and incubate at room temperature in the dark for 3 hours; ce...

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Abstract

The invention relates to a method for assembling a fluorescence detection device based on binding-induced DNA and detecting the content of small molecules. A combination-induced DNA assembly fluorescence detection device, including recognition elements and signal elements, the recognition elements include colloidal gold, colloidal gold surface-modified specific antibodies corresponding to the small molecules to be detected and a DNA sequence that is the auxiliary probe AO The signal element includes a DNA sequence, one end of the DNA sequence is marked with a fluorophore, the other end is connected to a small molecule antigen to be detected, a section of DNA sequence C1* at one end of the fluorophore is marked and a free end of the auxiliary probe AO A sequence of C1 is complementary, and the middle part of the signal element can be bent freely. The method of the invention is a homogeneous detection, which is simple and convenient, and can be applied to the detection of different small molecules by using different antibodies.

Description

technical field [0001] The invention belongs to the technical field of detection, and in particular relates to a method for assembling a fluorescent detection device based on binding-induced DNA and detecting the content of small molecules. Background technique [0002] Binding-induced DNA assembly technology refers to that when the recognition probe containing nucleotides specifically binds to the target molecule, a DNA assembly reaction is triggered and a fluorescent detection signal is generated. That is, DNA assembly can only be achieved when a specific target molecule elicits a specific binding reaction. This technique requires two affinity ligands to recognize the same target molecule, and is mostly used for the detection of macromolecules such as proteins and nucleic acids. The existing binding-induced DNA assembly technology is not suitable for small molecule detection, because small molecules cannot simultaneously provide two affinity ligands for specific binding r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577C12Q1/6804
CPCG01N33/577C12Q1/6804C12Q2563/107
Inventor 张兆威许琳李培武张奇张文姜俊
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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