Electrochemical biosensor for detecting ochratoxin A and preparation method of electrochemical biosensor

A biosensor and ochratoxin technology, applied in biochemical equipment and methods, material electrochemical variables, scientific instruments, etc., can solve the problems of complicated instrument operation and expensive equipment, and achieve good repeatability, low detection limit, and high speed fast effect

Active Publication Date: 2020-07-17
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims at the shortcomings of complex instrument operation, expensive equipment and professional operation in the existing detection method, and provides a preparation method of an electrochemical biosensor for detecting ochratoxin A with high sensitivity, strong specificity and low cost

Method used

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  • Electrochemical biosensor for detecting ochratoxin A and preparation method of electrochemical biosensor
  • Electrochemical biosensor for detecting ochratoxin A and preparation method of electrochemical biosensor
  • Electrochemical biosensor for detecting ochratoxin A and preparation method of electrochemical biosensor

Examples

Experimental program
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Effect test

Embodiment 1

[0064] A preparation method of the electrochemical biosensor of the present invention, comprising the following steps:

[0065] a. The gold electrode is first polished in 0.3 and 0.05 µm alumina slurry until it becomes a mirror surface, and rinsed repeatedly with PBS and secondary water;

[0066] b. Take 25 µL sterilized water, 10 µL 5×PBS buffer, 5 µL BK chain (2.5 µM), 5 µL S1 chain (2.5 µM), 5 µL ST chain (2.5 µM) in a sterile centrifuge tube, shake After 30 s, put it in a water bath at 95°C for 5 min, and then cool to room temperature;

[0067] c. Add 10 μL of the solution in step b to the surface of the electrode dropwise, incubate at room temperature for 2 h, and wash.

[0068] So far, the modification process of the electrode has come to an end. The following describes the reaction in the homogeneous solution and the main steps in the homogeneous reaction:

[0069] a. Add 5 µL aptamer (10 µM) and 5 µL T (10 µM) into a sterilized centrifuge tube, shake for 30 s, place ...

Embodiment 2

[0077] A preparation method of the electrochemical biosensor of the present invention, comprising the following steps:

[0078] a. The gold electrode is first polished in 0.3 and 0.05 µm alumina slurry until it becomes a mirror surface, and rinsed repeatedly with PBS and secondary water;

[0079] b. Take 25 µL sterilized water, 10 µL 5×PBS buffer, 5 µL BK chain (2.5 µM), 5 µL S1 chain (2.5 µM), 5 µL ST chain (2.5 µM) in a sterile centrifuge tube, shake After 30 s, put it in a water bath at 95°C for 5 min, and then cool to room temperature;

[0080] c. Add 10 μL of the solution in step b to the surface of the electrode dropwise, incubate at room temperature for 2 h, and wash.

[0081] So far, the modification process of the electrode has come to an end. The following describes the reaction in the homogeneous solution and the main steps in the homogeneous reaction:

[0082] a. Add 5 µL aptamer (10 µM) and 5 µL T (10 µM) into a sterilized centrifuge tube, shake for 30 s, place ...

Embodiment 3

[0090] A preparation method of the electrochemical biosensor of the present invention, comprising the following steps:

[0091] a. The gold electrode is first polished in 0.3 and 0.05 µm alumina slurry until it becomes a mirror surface, and rinsed repeatedly with PBS and secondary water;

[0092] b. Take 25 µL sterile water, 10 µL 5×PBS buffer, 5 µL BK chain (2.5 µM), 5 µL S1 chain (0.5 µM, 1 µM, 1.5 µM, 2 µM, 2.5 µM, 3 µM, 3.5 µM, 4 µM), 5 µL ST chain (2.5 µM) in a sterilized centrifuge tube, shaken for 30s, put in a 95°C water bath for 5 min, and then cool to room temperature;

[0093] c. Add 10 μL of the solution in step b to the surface of the electrode dropwise, incubate at room temperature for 2 h, and wash.

[0094] So far, the modification process of the electrode has come to an end. The following describes the reaction in the homogeneous solution and the main steps in the homogeneous reaction:

[0095] a. Add 5 µL aptamer (10 µM) and 5 µL T (10 µM) into a sterilized...

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Abstract

The invention relates to the technical field of biosensors, and in particular to an arched probe specifically recognized by ochratoxin A (OTA) and a biosensor for detecting OTA by catalytic hairpin assembly (CHA) and chain displacement isothermal cyclic amplification, and aims to solve the problems of low specificity and sensitivity and high cost of a method for detecting OTA in the prior art. According to the biosensor for detecting OTA, an electrode is sequentially modified with an ST-BK-S1 layer, an aptamer-T-HP1-HP2-HP3-F layer and a heme layer. A preparation method comprises the followingsteps: pretreating the electrode; modifying the surface of the electrode with the ST-BK-S1 layer; modifying the surface of the electrode with the aptamer-T-HP1-HP2-HP3-F layer; and modifying the surface of the electrode with the heme layer. High specificity detection of the target OTA is realized by utilizing specificity recognition of the OTA, and two-step amplification of the target signal is realized by utilizing a CHA reaction and the chain displacement isothermal amplification technology.

Description

technical field [0001] The invention relates to the technical field of electrochemical sensors, in particular to an electrochemical biosensor for detecting ochratoxin A and a preparation method thereof. Background technique [0002] Ochratoxin A (OTA) is an important mycotoxin secreted by Aspergillus and Penicillium on crops such as corn and legumes. It has kidney toxicity, liver toxicity, teratogenicity and carcinogenicity to animals and humans. and immunosuppressive effects. [0003] At present, there are many methods for detecting ochratoxin A, such as optical analysis, electrophoresis, chromatography and so on. However, these analytical methods have disadvantages such as cumbersome procedures, expensive instruments, and complicated equipment, and are not suitable for on-site detection. Therefore, developing a rapid and accurate method for OTA detection is of great significance for controlling food safety and quality. Contents of the invention [0004] The invention ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6825C12Q1/682G01N27/48G01N27/327
CPCC12Q1/6825C12Q1/682G01N27/48G01N27/3277C12Q2565/607C12Q2525/205C12Q2525/301Y02A50/30
Inventor 黄加栋王业茹王玉刘素孙文玉张曼茹江龙
Owner UNIV OF JINAN
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