Colorimetric detection method based on exonuclease signal amplification

一种核酸外切酶、信号放大的技术，应用在比色检测领域，能够解决吸光度响应小、灵敏度低等问题，达到合成方法简单、稳定性好、实现信号放大的效果

Pending Publication Date: 2021-01-15
YANTAI UNIV
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Researchers have reported using nucleic acid aptamers as recognition molecules to detect kanamycin without signal amplification. However, the absorbance response of this method is small and the sensing sensitivity is low (0.53nM).

Method used

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Embodiment

[0034] This embodiment is a colorimetric detection method based on exonuclease signal amplification. A partially complementary double-stranded DNA (dsDNA) probe is used as a recognition molecule. After the dsDNA probe interacts with the target, it becomes single-stranded DNA. Exo I can specifically hydrolyze the characteristics of single-stranded DNA, release the target molecule into the next round of reaction, and realize the signal amplification of the system. The process is shown in the appendix Image 6 shown.

[0035] The specific steps of the method are as follows:

[0036] In step 1, the dsDNA probe is used as a recognition molecule, and the dsDNA probe becomes single-stranded DNA after interacting with the target to be tested;

[0037] Step 2, utilize the characteristic of Exo I to specifically hydrolyze single-stranded DNA, release the target molecule to be tested into the next round of reaction, and then realize signal amplification;

[0038] Step 3, adding polycat...

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Abstract

The invention provides a colorimetric detection method based on exonuclease signal amplification. The method comprises the following steps: 1, reacting a dsDNA probe serving as a recognition moleculewith a target object to chang the dsDNA probe into single-stranded DNA; 2, releasing target molecules to enter the next round of reaction by utilizing the characteristic that the single-stranded DNA is specifically hydrolyzed by Exo I, so as to realize signal amplification; 3, adding polycations into the solution to react with the remaining dsDNA probe; and 4, adding negatively charged nanoparticles, and enabling the negatively charged nanoparticles to react with residual polycations in the system, so as to obtain the concentration of the target object. The method disclosed by the invention has the advantages that 1) the sensitivity is high; 2) naked eye real-time observation can be realized; 3) the method is simple, rapid, small in reaction volume, low in cost and suitable for conventional quality control of laboratories; and 4) the spectrum is broad: the content of different target objects in different samples can be detected.

Description

technical field [0001] The invention belongs to the field of colorimetric detection methods, in particular to a colorimetric detection method based on exonuclease signal amplification. Background technique [0002] Colorimetric technology has become a technology that has attracted much attention in the field of chemical and biological sensing. Nanoparticle-based colorimetric methods have been greatly developed due to their excellent localized surface plasmon resonances and high extinction coefficients. In particular, colorimetric methods based on the change of the existing state of nanoparticles, that is, from dispersion to aggregation, and color changes, are widely used in the detection and analysis of metal ions, DNA, small molecules, protein molecules, bacteria, etc. [0003] To obtain high sensitivity, different signal amplification techniques are used in aptamer-based colorimetric biosensors. Such as heterochain reaction (HCR), polymerase chain reaction (PCR), tool en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/58G01N33/569G01N33/68
CPCG01N33/5308G01N33/587G01N33/56983G01N33/56966G01N33/56961G01N33/56911G01N33/68
Inventor 付秀丽李婧闻刘永明
Owner YANTAI UNIV
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