Fluorescence enhanced type aptamer sensor as well as preparation method and application thereof

An aptamer sensor and fluorescence enhancement technology, applied in the field of biosensors, can solve the problems of high cost, harsh detection conditions, and high operational requirements, and achieve the effects of increased fluorescence radiation amplitude, high biocompatibility, and wide detection range.

Active Publication Date: 2018-11-23
ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention combines the high specificity selection of nucleic acid aptamers and the excellent characteristics of metal nanoparticles to provide a fluorescence-enhanced aptamer sensor, which overcomes the high operational requirements, high cost, and Disadvantages such as harsh detection conditions

Method used

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  • Fluorescence enhanced type aptamer sensor as well as preparation method and application thereof
  • Fluorescence enhanced type aptamer sensor as well as preparation method and application thereof
  • Fluorescence enhanced type aptamer sensor as well as preparation method and application thereof

Examples

Experimental program
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Embodiment 1

[0054] The detection of the optimal distance between embodiment 1 fluorophore and gold nanoparticle

[0055] In this embodiment, the preparation experiments of fluorescence-enhanced aptasensors are divided into 6 groups according to the difference in the distance between the fluorescent group and the gold nanoparticles.

[0056] The ATP nucleic acid aptamer used in group 1 is ATP nucleic acid aptamer 1, namely D1;

[0057] The ATP nucleic acid aptamer used in group 2 is ATP nucleic acid aptamer 1, namely D2;

[0058] The ATP nucleic acid aptamer used in group 3 is ATP nucleic acid aptamer 1, namely D3;

[0059] The ATP nucleic acid aptamer used in group 4 is ATP nucleic acid aptamer 1, namely D4;

[0060] The ATP nucleic acid aptamer used in group 5 is ATP nucleic acid aptamer 1, namely D5;

[0061] The ATP nucleic acid aptamer used in group 6 is ATP nucleic acid aptamer 1, namely D6.

[0062] The preparation of each group of fluorescence-enhanced aptasensors comprises the...

Embodiment 2

[0071] Example 2 The optimal volume ratio experiment of gold nanoparticle solution and ATP nucleic acid aptamer

[0072] In this example, according to the volume ratio of the gold nanoparticle solution (AuNPs) and the ATP nucleic acid aptamer, the fluorescence-enhanced aptamer sensor preparation experiment was divided into 4 groups, and each group used D1-D6 as the target nucleic acid aptamer .

[0073] In group 1, the volume ratio of gold nanoparticle solution (AuNPs) and ATP nucleic acid aptamer is 5:1;

[0074] In group 2, the volume ratio of gold nanoparticle solution (AuNPs) and ATP nucleic acid aptamer is 8:1;

[0075] In group 3, the volume ratio of gold nanoparticles solution (AuNPs) and ATP nucleic acid aptamer was 10:1,

[0076] In group 4, the volume ratio of gold nanoparticles solution (AuNPs) and ATP nucleic acid aptamer was 12:1.

[0077] The preparation of each group of fluorescence-enhanced aptasensors includes the following steps:

[0078] Step 1: Centrifu...

Embodiment 3

[0093] 1) Preparation of fluorescence-enhanced aptasensor, comprising the following steps:

[0094] Step 1: Centrifuge the ATP aptamer D3 at 12000 rpm for 1 min, then dilute it to 5 μmol / L with TCEP, and activate it for 2 h at room temperature in the dark without light; mix 800 μL of gold nanoparticle solution (AuNPs) and 100 μL of ATP aptamer D3 was mixed and incubated for 18h.

[0095] Step 2: Add 100 μL of 1×PBS to the solution obtained in step 1, and react for 6 hours. The function of 1×PBS is to adjust the pH value and increase the ionic strength of the solution.

[0096] Step 3: Add 2 mol / L NaCl solution to the solution obtained in Step 2, repeat every 3 hours, repeat 2 times, so that the concentration of NaCl in the total solution can gradually increase to 0.2 mol / L, and let it stand for 24 hours.

[0097] Step 4: Centrifuge the solution obtained in step 3 at 14°C and 15,000 rpm for 15 minutes, centrifuge three times in total; remove the supernatant, dissolve the obtai...

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Abstract

The invention belongs to the technical field of biosensors and specifically relates to a fluorescence enhanced type aptamer sensor as well as a preparation method and application thereof. A metal nanostructure is guided into a fluorescent molecular detection system, aptamer for modifying a certain amount of basic groups A is combined with metal nanoparticles through sulfydryl, and plasma resonancegenerated by the metal nanoparticles is utilized to improve fluorophore excitation efficiency; thus, fluorescent radiation amplitude is remarkably improved, and the purpose of amplifying fluorescencesignals is achieved. Meanwhile, the fluorescence enhanced type aptamer sensor which is used for detecting low-concentration target objects is provided by being combined with specificity selection ofaptamer, so that flexibility, accuracy and specificity of analysis detection are ensured, and detection of polyphyly of the target object is also achieved. Meanwhile, the fluorescence enhanced type aptamer sensor has the characteristics of low cost, no pollution and wider detection range.

Description

technical field [0001] The invention belongs to the technical field of biosensors, in particular to a fluorescence-enhanced aptamer sensor and its preparation method and application. Background technique [0002] In the fields of clinical diagnosis, food safety and environmental protection, the requirements for highly sensitive detection of biomolecules are getting higher and higher, and the construction of more sensitive and accurate sensing and detection methods has become the focus and hotspot of research in the field of life analysis science. Fluorescent aptamer sensor is a fluorescent sensor constructed by using nucleic acid aptamer as a recognition group. This sensor has the advantages of both fluorescence analysis and nucleic acid aptamer. It can perform quantitative analysis on a variety of biological small molecules. Broad prospects. However, the concentration of biomolecules in the actual environment is generally very low (such as insulin, dopamine, aflatoxin, etc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/53G01N33/533
CPCG01N21/6428G01N33/5308G01N33/533G01N2021/6432G01N2021/6439
Inventor 任林娇姜利英张培王慰张吉涛曹玲芝姜素霞王延峰
Owner ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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