Preparation method of unmarked ratio electrochemical sensor for detecting aflatoxin B1 based on time regulation sensitivity

A kind of aflatoxin and label-free technology, which is applied in the field of biosensors, can solve the problems of different degrees of mildew, and the amount of AFB1 is not in the same order of magnitude, so as to achieve the effect of improving selectivity, high sensitivity and good selectivity

Inactive Publication Date: 2020-08-14
JIANGSU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, the degree of mildew of crops due to lack of space or the influence of weather during harvesting, storage, transportation, etc. is different, so the amount of AFB1 produced is not in the same order of magnitude

Method used

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  • Preparation method of unmarked ratio electrochemical sensor for detecting aflatoxin B1 based on time regulation sensitivity
  • Preparation method of unmarked ratio electrochemical sensor for detecting aflatoxin B1 based on time regulation sensitivity
  • Preparation method of unmarked ratio electrochemical sensor for detecting aflatoxin B1 based on time regulation sensitivity

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The preparation method of the ratiometric electrochemical biosensor comprises the following steps:

[0035] (1) Preparation of THI-rGO composites:

[0036] Weigh 1mg THI and dissolve it in ultrapure water to obtain a concentration of 20μg·mL in 50mL -1 THI solution, and serially diluted to 5 μg·mL -1 , stored away from light at room temperature; weigh 3mg rGO and disperse it in 30mL ultrapure water to obtain a concentration of 0.1mg·mL -1 The rGO dispersion solution was treated with a cell pulverizer for 30 min and stored at room temperature until use.

[0037] Take 3mL 5μg·mL -1 THI with 15mL 0.1mg·mL -1 The rGO was mixed and stirred in a 100mL beaker for 12h. After the stirring was completed, the solution was placed in a 10mL centrifuge tube, and was centrifuged and washed with water at a rotation speed of 8000rpm and a centrifugation time of 15min to obtain a precipitate. The obtained precipitate was dissolved in 3 mL ultrapure water, and stored at room temperat...

Embodiment 2

[0045] Adjust the interaction time between the Fc-modified AFB1 aptamer and the cDNA of the sensing interface and obtain sensors with different ratio signals:

[0046] The incubation time of the Fc-modified AFB1 aptamer at the sensing interface in step (6) of Example 1 is regulated, such as figure 1 As shown, the Fc-modified AFB1 aptamer incubation time was 80min (a), 100min (b), 120min (c) of the ratiometric sensor.

Embodiment 3

[0048] The sensor obtained by regulating and controlling in embodiment 2 is carried out ACV test, the electrochemical signal I of three sensors of gained THI with I Fc Such as figure 2 As shown in A.

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Abstract

The invention belongs to the technical field of biosensors, and particularly relates to a preparation method of an unmarked ratio electrochemical sensor for detecting aflatoxin B1 based on time regulation sensitivity. The invention discloses a label-free signal probe-thionine and reduced graphene oxide (THI-rGO) composite material. The composite material is formed through non-covalent interactionof thionine (THI) and reduced graphene oxide (rGO). The gold nanoparticles (AuNPs) fix sulfydryl modified aptamer complementary chains on a sensing interface through Au-S covalent bonds; and the aptamers marked by the electrochemical active molecules Fc at the 3'and 5' ends are fixed on a modified sensing interface through the complementary pairing effect of basic groups, so that the novel sensoris constructed. Furthermore, the ratio (ITHI/IFc) of ratio signals is regulated and controlled by regulating and controlling the cDNA action time of the Fc-modified AFB1 aptamer and a sensing interface, so that the novel sensitivity-controllable label-free ratio electrochemical biosensor is obtained and is used for sensitively and quickly analyzing an actual sample AFB1.

Description

technical field [0001] The invention belongs to the technical field of biosensors, and in particular relates to a preparation method and application of a label-free ratiometric electrochemical biosensor based on adjustable sensitivity, which can be used for detecting aflatoxin B1 (AFB1). Background technique [0002] AFB1 is the most widespread and most toxic type of mycotoxin in crops. It is highly resistant to degradation. It can not only survive in food processing, but also damage the liver of animals or humans through the food chain, and even cause cancer. Therefore, the accurate detection of AFB1 is of great significance to ensure the quality and safety of crops and protect human health. [0003] At present, the analysis methods for AFB1 mainly include enzyme-linked immunosorbent assay (ELISA), thin-layer chromatography (TLC), and liquid-mass spectrometry (HPLC-MS). These methods are widely used with their own unique advantages and have achieved good detection results....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3278
Inventor 由天艳李玉叶刘东李文佳朱成喜
Owner JIANGSU UNIV
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