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Target object and redox dual-response aptamer sensor, preparation method and application thereof, and quantitative detection method of anabaena toxins

An aptamer sensor and quantitative detection technology, which can be applied to electrochemical variables of materials, analysis by chemical reaction of materials, chemiluminescence/bioluminescence, etc. It can solve the problem of electrochemiluminescence biosensors without anabaena toxins question

Active Publication Date: 2021-09-03
ANHUI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The analysis of anabatoxin-dependent metal ion-dependent DNAzyme electrochemiluminescent nano-hybrid systems has little research, and there is no electrochemiluminescent biosensor for detecting anabaena toxins in the prior art

Method used

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  • Target object and redox dual-response aptamer sensor, preparation method and application thereof, and quantitative detection method of anabaena toxins
  • Target object and redox dual-response aptamer sensor, preparation method and application thereof, and quantitative detection method of anabaena toxins
  • Target object and redox dual-response aptamer sensor, preparation method and application thereof, and quantitative detection method of anabaena toxins

Examples

Experimental program
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Effect test

Embodiment 1

[0056] A method for preparing a target substance and a redox dual-response aptamer sensor, comprising the following steps:

[0057] (1) Disperse the AgNPs@Ru-MOF material in deionized water to prepare a AgNPs@Ru-MOF solution with a concentration of 1 mg / mL, mix and incubate 20 μL AgNPs@Ru-MOF solution with 20 μL 1 μM DNA S1 solution for 12 h to obtain S1- AgNPs@Ru-MOF solution;

[0058] (2) Polish the GCE electrode into a mirror surface with alumina slurry, clean it with ultrapure water, and dry it with nitrogen; place the polished GCE electrode in 2mL 1mM ABA solution, use silver / silver chloride as the reference electrode, Platinum wire was used as the counter electrode, and electrodeposition was carried out at a scanning potential of 0.4-1.2V at 10mV / s. After three cycles, carboxy-modified ABA / GCE was obtained, and then placed in 1mL of a mixed solution containing equimolar EDC and NHS for 2h , the concentration of EDC in the mixed solution is 5mM, washed with ethanol and u...

Embodiment 2

[0064] A method for quantitative detection of anabatoxin, comprising the following steps:

[0065] A, using the preparation method in Example 1 to prepare multiple groups of aptasensors;

[0066] B. Put each ligand sensor in step A into 10 μL of 0.001mg / mL, 0.01mg / mL, 0.1mg / mL, 0.2mg / mL, 0.4mg / mL, 0.6mg / mL, 0.8mg / mL, Soak in 1.0mg / mL concentration of anabatoxin aqueous solution for at least 2.0h, and then wash with 10mM Tris-HCl buffer solution with pH=7.4;

[0067] C. Put each electrochemiluminescent biosensor into 1mL 0.1mol / L hydrogen peroxide aqueous solution and soak for at least 2.0h, then take it out and dry it;

[0068] D. Use the electrochemiluminescent biosensors treated in step C as the working electrode, silver / silver chloride as the reference electrode, and platinum wire as the counter electrode to form a three-electrode system. Use an MPI-E ECL analyzer at 8 mL pH The electrochemiluminescent signal intensity of each electrode system is tested in the 0.1M PBS bu...

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Abstract

The invention discloses a target object and redox dual-response aptamer sensor, a preparation method and application thereof and a quantitative detection method of anabaena toxins. DNA S1 is connected to an AgNPs@Ru-MOF material through an Ag-S bond; a solution of the material, a DNA S3 solution and a solution obtained after mixed incubation of DNA S2 and ferrocenecarboxylic acid are sequentially modified to the surface of a pretreated GCE electrode to prepare the aptamer sensor, when a target object anabaena toxin exists, a second structure of DNA S3 can be triggered, so that DNA S1 and DNA S2 form a special structure capable of being recognized by silver ions, and then along with addition of H2O2, H2O2 can etch silver nanoparticles to generate silver ions, the silver ions cut DNA S2 by recognizing cutting sites to release an inhibitor ferrocene, electrochemical luminescence of ruthenium on the surface of the electrode is induced, and quantitative detection of anabaena toxins is achieved.

Description

technical field [0001] The invention belongs to the technical field of biosensors, and relates to a target object and a redox dual-response aptamer sensor and its preparation method and application, as well as a quantitative detection method for anabatoxins, in particular to an ECL-RET-based AgNPs@Ru -The target of MOF probe and redox dual response aptamer sensor, its preparation method and application, and the quantitative detection method of anabatoxin. Background technique [0002] Anatoxin-a (ATX-a, anabatoxin) is the smallest neurotoxin (Mol.Wt, 165.23Da), an alkaloid that affects the normal release of acetylcholine and causes neuromuscular hyperexcitability and convulsions, Ultimately, respiratory restriction and suffocation result. So far, many analytical methods for anabatoxins have been reported. For example, the group of Samuel P. Haddad used isotope dilution liquid chromatography tandem mass spectrometry for the determination of anabatoxins. AzevedoJ' group dev...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N27/30G01N27/26
CPCG01N21/76G01N27/30G01N27/26Y02A50/30
Inventor 王广凤夏蒙蒙
Owner ANHUI NORMAL UNIV
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