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Preparation method of electrochemical adapter sensor for detecting fumonisin B1 ( FB1)

An aptamer sensor and fumonisin technology, which is applied in the field of electrochemical detection, can solve the problems that antibodies are susceptible to external environment interference, high preparation cost, and difficult to store, and achieves avoiding preprocessing procedures, high sensitivity, and reagent dosage. less effect

Inactive Publication Date: 2017-05-24
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method uses antibodies as the recognition elements, but the preparation of antibodies requires animal experiments or cell experiments, which is cumbersome and time-consuming, and the preparation cost is high. Moreover, the obtained antibodies are easily disturbed by the external environment and difficult to store. These shortcomings limit the popularization and application of this method.

Method used

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  • Preparation method of electrochemical adapter sensor for detecting fumonisin B1 ( FB1)

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Experimental program
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Effect test

Embodiment 1

[0031] SPCE preprocessing

[0032] (1) SPCE is cleaned: the processing method is in 5mL H 2 SO 4 (0.5M) SPCE was scanned by cyclic voltammetry in a solution containing 0.1M KCl, the scan range was set at -0.2 to 1.5V, and the scan rate was 100mV s -1 , until the overlapping cyclic voltammetry curves are obtained by scanning, rinse with water twice, dry with nitrogen, and set aside.

Embodiment 2

[0034] Activation of aptamers

[0035] (2) Preparation of activated FB1 aptamer solution: dissolve the FB1 aptamer with 50 mM Tris-HCl buffer solution to make the concentration of the FB1 aptamer 100 μM to obtain solution A; take 20 μL of solution A and add it to 5 μL In 50mM Tris-HCl buffer solution B (pH=7.4) containing 100mM tris (2-carboxyethyl) phosphine hydrochloride (TCEP), the mixed solution C was obtained; then, the mixed solution C was placed on a reciprocating shaker React at room temperature for 1 h; finally, centrifuge to remove excess tris(2-carboxyethyl)phosphine hydrochloride, redisperse in 2 mL of 50 mM Tris-HCl buffer solution D (pH=7.4, containing 0.1M NaCl, 0.2M KCl , 5.0 mM MgCl 2 and 1.0mM EDTA), the activated FB1 nucleic acid aptamer solution was obtained for subsequent use.

Embodiment 3

[0037] Construction of label-free electrochemical aptasensor

[0038] (3) Construction of AuNPs-SPCE: Immerse the above treated SPCE into 5mL HAuCl 4 Solution (2mM) was used to deposit gold nanoparticles on the surface, the deposition voltage was 0.5V, and the deposition time was set to 1200s to obtain AuNPs-SPCE, rinsed with double distilled water, dried with nitrogen, and set aside.

[0039] (4) The relationship between the amount of electrodeposited AuNPs and the deposition time: set different deposition times, conduct electrodeposition on the SPCE surface, perform electrochemical impedance analysis on the electrodes under different deposition times, obtain a series of electrochemical impedance spectra, and Plot the time-impedance relationship curve.

[0040] (5) Preparation of AuNPs-SPCE modified by FB1 nucleic acid aptamer: First, take 10 μL of activated FB1 nucleic acid aptamer solution and drop-coat it on the working electrode surface of the electrode obtained in (3), ...

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Abstract

The invention provides a preparation method of an electrochemical adapter sensor for detecting fumonisin B1 (FB1). The preparation method comprises the following steps of step one, preparation of an silk-printed carbon electrode (SPCE); step two, preparation of an activated FB1 aptamer solution; step three, preparation of an Au nanoparticles (AuNPs) modified SPCE; step four, preparation of an FB1 aptamer modified AuNPs-SPCE; step five, sealing of spare active sites on the surface of the electrode. According to the preparation method of the electrochemical adapter sensor for detecting the FB1, provided by the invention, the AuNPs are adopted for modifying the surface of the electrode, on one hand, the large amount of AuNPs are loaded on the surface of the electrode, so that a signal amplification function on label-free impedance method detection is obtained; on the other hand, more sulfhydrylated adapters are fixed to the surface of the electrode by utilizing Au-S bonds. An FB1 impedance detection method provided by the invention has the characteristics that the operation is more simple, convenient and flexible, instruments and equipment are simpler, the dosage of a reagent is less, the detection cost is low, and the like.

Description

technical field [0001] The invention belongs to the field of electrochemical detection, in particular to a construction method of a label-free electrochemical aptamer sensor for sensitive detection of fumonisin B1 in corn. Background technique [0002] Fumonisin B (FB) is a group of important mycotoxins produced by Fusarium moniliforme. It is a class of diester compounds with similar structures composed of different polyhydric alcohols and tricarboxylic acids. Fumonisins mainly exist in grains, most of which are corn and corn-based food and feed. Because the toxicity of fumonisins is species-specific and target-organ specific, the ingestion of FB will not only pose a potential threat to human and animal health but also cause damage to the brain, liver, lung and kidney to varying degrees. Existing data show that FB can cause equine encephalomalacia, pig pulmonary edema and primary liver cancer in rats, and cause immunosuppressive effects on poultry. In addition, studies hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/30G01N27/327C25D3/48C25D5/54B82Y30/00
CPCG01N27/308B82Y30/00C25D3/48C25D5/54G01N27/3278
Inventor 钱静任婵婵王成全徐丽王坤李华明
Owner JIANGSU UNIV
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