Pathogenic bacteria electrochemical detection method based on DNA walker and nanoflower structure

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

Active Publication Date: 2021-03-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The technology described by this patented allows for modification or enhanced activity at an elec-optical interface that can detect small amounts of substances (such as nucleic acids) without being affected negatively affecting their ability to perform its function effectively. This results from modifying certain parts of these molecules called DNA walks with specific sequences attached thereto. These modifications help them bind specifically to other molecular structures inside cells while still allowing some signals like those involved in gene expression to be detected accurately through fluorescent labelings placed near the elecrochromism layer.

Problems solved by technology

In this patented technical problem addressed in the patents relates to improving the accuracy and reproduciblence rates of detecting antibodysceptic staphylcytoxins (AS), particularly AS caused by Bacillustocholaceanis fragility strain B1(BFT). Current techniques like X-ray fluorescences analysis require long periods of exposure while maintaining their quality. However these techniques may result in false positives when analyzing samples containing other types of substances. There is thus a demand for more sensitive and reliable assays capable of quickly identifying AS without requiring lengthy steps and costly purifications.

Method used

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  • Pathogenic bacteria electrochemical detection method based on DNA walker and nanoflower structure
  • Pathogenic bacteria electrochemical detection method based on DNA walker and nanoflower structure
  • Pathogenic bacteria electrochemical detection method based on DNA walker and nanoflower structure

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Experimental program
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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 a pathogenic bacteria electrochemical detection method based on a DNA walker and a nanoflower structure, and belongs to the technical field of detection. A DNA walker is modified on the surface of the electrode, when a target object exists, a large number of free rolling circle amplification primers are released, formation of DNA nanoflowers is induced through a subsequentrolling circle amplification reaction, the effective area of the surface of the electrode is increased, a detection signal is amplified, and the detection sensitivity is effectively improved. And whenno target object exists in the solution, the DNA walker hydrolyzes the rolling circle amplification primer under the induction of exonuclease III, thereby reducing the background signal and wideningthe detection range of the sensor.

Description

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Claims

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Application Information

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Owner JIANGNAN UNIV
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