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Nucleic acid sensor based on quantum dots and preparation method and detection method thereof

A technology of nucleic acid sensor and detection method, which is applied in the field of biosensors, can solve the problems that have not yet been reported on nucleic acid sensor research, and achieve the effect of multiple repeated detection synchronization, good application prospects, and rapid detection

Inactive Publication Date: 2009-09-02
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to these unique quantum advantages, in recent years, the research on the application of quantum dots to FRET has been deepened and the scope has been continuously expanded, but so far, there has been no research report on the use of quantum dots as a nucleic acid sensor based on a solid support

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  • Nucleic acid sensor based on quantum dots and preparation method and detection method thereof
  • Nucleic acid sensor based on quantum dots and preparation method and detection method thereof
  • Nucleic acid sensor based on quantum dots and preparation method and detection method thereof

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preparation example Construction

[0047] Schematic diagram of the preparation and detection of the first quantum dot-based nucleic acid sensor figure 1 As shown, the preparation of the nucleic acid sensor is to firstly couple the oligonucleotide without self-complementary sequence to the quantum dot through the biotin-avidin (avidin or streptavidin) system to prepare one end Quantum dot-labeled oligonucleotide probes; then quantum dots and solid phase supports (for nucleic acid sensors, optional solid phase supports include silicon wafers, glass slides, tiles, polypropylene membranes or nylon membranes, etc.) ) is coupled through a biotin-avidin (avidin or streptavidin) system, thereby immobilizing the oligonucleotide probes on the surface of the solid phase support through quantum dots; using the same method to immobilize multiple oligonucleotide probes to form an oligonucleotide probe array; when the nucleic acid sensor is used for detection, one end of the nucleic acid sample to be tested is labeled with a ...

Embodiment 1

[0054] 1. Preparation of nucleic acid sensor

[0055] 1. Preparation of streptavidin-labeled quantum dots

[0056] (1) Preparation of quantum dots

[0057] Mix sodium sulfate and selenium powder in a molar ratio of 3:1, add 150 mL of deionized water, and reflux reaction under nitrogen protection conditions until the selenium powder is completely dissolved to obtain a clear sodium selenosulfate solution; 0.46 g of cadmium chloride Dissolve in 240mL of deionized water, stir and react for 30 minutes under nitrogen protection, then add 0.36mL of thioglycolic acid dropwise, adjust the pH value of the solution to 11.2 with NaOH with a concentration of 1mol / L, continue to stir and react for 30 minutes, then drop Add 12 mL of the above-mentioned sodium selenosulfate solution, and by controlling the time of the reflux reaction, a CdSe quantum dot solution with an emission wavelength of 492 nm and a carboxyl group modified on the surface is prepared.

[0058] The UV-Vis absorption spe...

Embodiment 2

[0075] 1. Preparation of nucleic acid sensor

[0076] 1. Preparation of streptavidin-labeled quantum dots

[0077] (1) Preparation of quantum dots

[0078] Na 2 SeSO 3 Solution preparation: In a 250mL three-necked bottle, add Na 2 SO 3 6.302g (0.05mol) and Se powder 1.579g (0.02mol), add high-purity water 100mL to dissolve, stirring, N 2 Under protective conditions, heat up to 90°C for reflux reaction. As the reaction time prolongs, the solution containing gray-black precipitate gradually becomes clear and transparent. After 6 hours of reaction, stop heating and filter with suction to obtain a colorless and transparent filtrate, that is, the concentration is about 0.2mol / L. Na 2 SeSO 3 The solution is stored at a temperature of 70°C (to prevent oxidation) and is used for later use.

[0079] Preparation of NaHSe solution: In a 20mL small flask, add 0.1895g (0.0024mol) of Se powder, add 6mL of high-purity water to dissolve, and then add newly prepared NaBH 4 Solution [...

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Abstract

The invention discloses a nucleic acid sensor based on quantum dots and a preparation method and a detection method thereof, wherein the nucleic acid sensor comprises a solid-phase holder and an oligonucleotide probe array fixed on the surface of the solid phase holder, wherein the oligonucleotide probe array comprises an oligonucleotide probe which does not contain self-complementary sequences; one end of the oligonucleotide probe marks quantum dots and is fixed on the surface of the solid-phase holder by the quantum dots; or the oligonucleotide probe array comprises a molecular beacon, wherein one end of the molecular beacon marks quantum dots and is fixed on the surface of the solid-phase holder by the quantum dots, and the other end of the molecular beacon is marked by a fluorescence quenching group. By utilizing the basic-group complementation pairing principle and the FRET phenomenon, the detection of the special nucleic acid sequences in a nucleic acid sample can be simultaneously achieved; in addition, the invention has simple preparation and accurate, sensitive, simple, convenient and rapid detection and can detect a plurality of samples simultaneously.

Description

technical field [0001] The invention relates to a biosensor, in particular to a nucleic acid sensor based on quantum dots, and also relates to a preparation method and a detection method of the nucleic acid sensor. Background technique [0002] In the field of nucleic acid analysis, as an accurate, simple and rapid detection method, nucleic acid sensors have received more and more attention. The basic principle is to immobilize oligonucleotide probes with labels that can recognize a specific nucleic acid sequence on a solid support, perform a hybridization reaction with the nucleic acid sample to be tested, and convert the hybridization reaction results into readable The obtained signal is output, so as to realize the qualitative or quantitative detection of the specific nucleic acid sequence in the nucleic acid sample to be tested. [0003] The output signal of the nucleic acid sensor during detection can be an electrical signal or an optical signal, etc., depending on the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 罗阳府伟灵王珏
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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