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Aptamer sensor based on nano particles and chemiluminiscence, as well as preparation method and application of sensor

A nucleic acid aptamer, chemiluminescence technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, etc., can solve the problems of cumbersome separation operation, inconvenient chemiluminescence signal amplification, detection speed and sensitivity impact, etc. To achieve the effect of broad application prospects, long luminous time and wide application range

Active Publication Date: 2014-04-23
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The separation operation of the existing chemiluminescent nucleic acid aptamer sensor is relatively cumbersome, and it is not convenient to amplify the chemiluminescent signal, which leads to a certain impact on the detection speed and sensitivity

Method used

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  • Aptamer sensor based on nano particles and chemiluminiscence, as well as preparation method and application of sensor
  • Aptamer sensor based on nano particles and chemiluminiscence, as well as preparation method and application of sensor
  • Aptamer sensor based on nano particles and chemiluminiscence, as well as preparation method and application of sensor

Examples

Experimental program
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preparation example Construction

[0031] The preparation method of the above-mentioned nucleic acid aptamer sensor based on nanoparticles and chemiluminescence uses magnetic nanoparticles and gold nanoparticles as a solid phase carrier, and uses chemiluminescence as a detection means, comprising the following steps:

[0032] Step 1. Prepare Fe with a particle size of about 500 nm by soft template method 3 o 4 Magnetic nanoparticles, via seed polymerization and classical Coated a layer of silicon shell on its surface to prepare Fe with a particle size of about 550nm 3 o 4 SiO 2 magnetic nanoparticles, and then carboxylated their surface to obtain carboxylated Fe 3 o 4 SiO 2 Magnetic nanoparticles, stored in absolute ethanol. Carboxylated Fe 3 o 4 SiO 2 Magnetic nanoparticles require carboxyl activation before use.

[0033] The Fe 3 o 4 The preparation methods of magnetic nanoparticles include co-precipitation method, solvothermal method, soft template method, microemulsion method, thermal decomposi...

Embodiment 1

[0045] The carboxylated Fe of embodiment 1 activation 3 o 4 SiO 2 Preparation of Magnetic Nanoparticles

[0046] Fe 3 o 4 Magnetic nanoparticles were prepared by soft template method, Fe 3 o 4 SiO 2 Magnetic nanoparticles using seed polymerization and classical Preparation method; the specific preparation process is carried out with reference to Example 1 of the invention application CN102568728A. The prepared particles have the characteristics of uniform size and good dispersibility; and then carry out carboxylation modification on the surface.

[0047] The specific experimental process is as follows.

[0048] (1), first adopt the soft template method to prepare Fe with a particle size of about 500nm 3 o 4 Magnetic nanoparticles, followed by seed polymerization and classical Coated a layer of silicon shell on its surface to prepare Fe with a particle size of about 550nm 3 o 4 SiO 2 magnetic nanoparticles. figure 1 gives Fe 3 o 4 SiO 2 Transmission electro...

Embodiment 2

[0055] Example 2 Preparation of bifunctional gold nanoparticles

[0056] First, the sodium citrate reduction method was used to prepare gold nanoparticles, and then the gold nanoparticles were combined with two different thiol-modified DNAs to prepare bifunctional gold nanoparticles.

[0057] The specific experimental process is as follows.

[0058] (1) Gold nanoparticles were prepared by sodium citrate reduction method. First add 48ml of double distilled water to the flask with condenser tube, then add 2ml of 1wt% HAuCl 4 ·3H 2 O solution, heated to boiling under vigorous stirring, and the stirring speed was 750r / min; after boiling for 5 minutes, quickly add 5ml of 38.8mmol / L sodium citrate solution; after discoloration, continue to reflux for 15 minutes, then remove the heating equipment, and naturally cool to room temperature, Store at 4°C. figure 2 The transmission electron microscope image of gold nanoparticles is given, and it can be seen that the particles are sphe...

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Abstract

The invention discloses an aptamer sensor based on nano particles and chemiluminiscence. The aptamer sensor takes Fe3O4@SiO2 magnetic nano particles and double-functional gold nano particles as solid-phase carriers; activated carboxylated Fe3O4@SiO2 magnetic nano particles and an amino modified aptamer are combined to form a magnetic nano particle-aptamer compound; then the magnetic nano particle-aptamer compound and the double-functional gold nano particles are subjected to hybridization to form a magnetic nano particle-aptamer-double-functional gold nano particle compound; finally, the magnetic nano particle-aptamer-double-functional gold nano particle compound is combined with streptavidin marked alkaline phosphatase to obtain the sensor. The invention further discloses a preparation method and an application of the sensor. The aptamer sensor has the magnetism and is convenient in rapid magnetic separation, also has the advantages of being rapid by adopting a chemiluminiscence method and efficient in detection, and has a very great application value in the detection aspect of biomedicines.

Description

technical field [0001] The invention relates to the technical field of nucleic acid aptamer sensors, in particular to a nucleic acid aptamer sensor using nanoparticles as a solid phase carrier and chemiluminescence as a detection means, as well as its preparation method and application. Background technique [0002] Magnetic nanoparticles have excellent physical and chemical properties and biocompatibility, such as fast separation speed, high efficiency, simple operation, easy functionalization, easy automation, and no influence on the activity of separated substances. They have been widely used in the separation of cells. Sorting, separation, purification and detection of biological macromolecules, targeted diagnosis and treatment and many other fields. Among them, the detection of biomacromolecules by using magnetic nanoparticles is mainly to use them to specifically detect target molecules after the surface is modified with specific probes or antibodies. [0003] Gold na...

Claims

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Application Information

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IPC IPC(8): G01N21/76
Inventor 何农跃习志江
Owner SOUTHEAST UNIV
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