Electrochemical detection method of pathogenic bacteria based on DNA walker and nanoflower structure

A detection method, a technology of pathogenic bacteria, applied in the detection field, can solve the problem of fewer DNA nanoflowers, achieve the effect of increasing the effective area, reducing the background signal, and broadening the detection range

Active Publication Date: 2021-11-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, DNA nanoflowers are rarely used in chemical detection

Method used

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  • Electrochemical detection method of pathogenic bacteria based on DNA walker and nanoflower structure
  • Electrochemical detection method of pathogenic bacteria based on DNA walker and nanoflower structure
  • Electrochemical detection method of pathogenic bacteria based on DNA walker and nanoflower structure

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: the drafting of staphylococcus aureus concentration standard curve

[0040] with 30% H 2 o 2 and H 2 SO 4 Prepare the piranha solution, soak the gold electrode in the piranha solution for 15 minutes to remove surface impurities, clean the electrode surface and transfer it to ultrapure water for 5 minutes of ultrasonication, and then the electrode is cleaned with 0.5mol L -1 h 2 SO 4 Perform electrochemical cleaning and electrode activation, clean the electrode again with ultrapure water, and dry it with nitrogen. 0.2μmol·L -1 DNA walker sequence and 0.4 μmol L -1 Add the aptamer to PBS buffer, 1 μmol L -1 Rolling circle amplification primers and 2 μmol L -1 The auxiliary sequence was added to PBS buffer, and the two mixtures were heat denatured at 95°C for 10 minutes, and then placed in an ice bath at 4°C for 10 minutes to form aptamer / DNA walker double strands and auxiliary sequence / rolling circle amplification primer pairs respectively. chain. ...

Embodiment 2

[0046] Embodiment 2: the mensuration of Staphylococcus aureus content in actual sample

[0047] In order to further verify the accuracy of this method in the determination of the content of Staphylococcus aureus in actual samples, Taihu Lake water and tap water without pretreatment and honey water diluted 5 times with PBS buffer were selected for the addition of Staphylococcus aureus Determination.

[0048] with 30% H 2 o 2 and H 2 SO 4 Prepare the piranha solution, soak the gold electrode in the piranha solution for 15 minutes to remove surface impurities, clean the electrode surface and transfer it to ultrapure water for 5 minutes of ultrasonication, and then the electrode is cleaned with 0.5mol L -1 h 2 SO 4 Perform electrochemical cleaning and electrode activation, clean the electrode again with ultrapure water, and dry it with nitrogen. 0.2μmol·L -1 DNA walker sequence and 0.4 μmol L -1 Add the aptamer to PBS buffer, 1 μmol L -1 Rolling circle amplification prim...

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Abstract

The invention discloses an electrochemical detection method for pathogenic bacteria based on a DNA walker and a nanoflower structure, and belongs to the technical field of detection. In the present invention, by modifying the DNA walker on the surface of the electrode, a large number of free rolling circle amplification primers are released when the target exists, and the subsequent rolling circle amplification reaction induces the formation of DNA nanoflowers, increases the effective area of ​​the electrode surface, and amplifies the detection signal. Effectively improve the detection sensitivity. When there is no target in the solution, the DNA walker hydrolyzes the rolling circle amplification primer under the induction of exonuclease III, which reduces the background signal and broadens the detection range of the sensor.

Description

technical field [0001] The invention relates to an electrochemical detection method for pathogenic bacteria based on a DNA walker and a nanoflower structure, and belongs to the technical field of detection. Background technique [0002] Pathogenic bacteria are microorganisms that can cause disease, also known as pathogenic microorganisms. Among them, Staphylococcus aureus (S.aureus) is a food-borne pathogen widely present in the environment. Enterotoxin produced by Staphylococcus aureus can cause food poisoning and various infections. The traditional detection method of Staphylococcus aureus has high sensitivity and specificity, but there are shortcomings such as time-consuming and complicated operation. Therefore, it is urgent to construct an accurate and simple detection method for Staphylococcus aureus. [0003] As a new detection method, biosensor has high sensitivity and high specificity, and is widely used in disease diagnosis, food safety and environmental monitori...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/26G01N27/30G01N27/327
CPCG01N27/26G01N27/30G01N27/3278
Inventor 周楠迪蔡蓉凤田亚平
Owner JIANGNAN UNIV
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