Electrochemical nucleic acid detection method based on DNA (deoxyribonucleic acid) walking and rolling circle amplification signal magnification

A detection method and nucleic acid technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of insufficient sensitivity, high detection cost, inconvenient operation, etc., and achieve low detection cost, high sensitivity, and easy operation Effect

Active Publication Date: 2020-05-19
天津国科医疗科技发展有限公司 +1
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  • Abstract
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However, there are still problems such as insufficient ...

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  • Electrochemical nucleic acid detection method based on DNA (deoxyribonucleic acid) walking and rolling circle amplification signal magnification
  • Electrochemical nucleic acid detection method based on DNA (deoxyribonucleic acid) walking and rolling circle amplification signal magnification
  • Electrochemical nucleic acid detection method based on DNA (deoxyribonucleic acid) walking and rolling circle amplification signal magnification

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Embodiment

[0030] Embodiment: Taking GCTAGAGATTTTCCACACTGACT as the target nucleic acid as an example, the DNA sequence involved in this embodiment is shown in Table 1:

[0031] Table 1 DNA sequence

[0032]

[0033] The detection principle of this case is as follows figure 1 Shown: First, silver nanoparticles were synthesized as electrochemical signal probes, which can be labeled on DNA chains modified with amino groups at the ends. Then two kinds of DNA tetrahedrons (DNA walker, DNA track) were assembled respectively, and fixed on the surface of the gold electrode according to a certain concentration ratio. This kind of tetrahedral DNA not only improves the molecular recognition efficiency of DNA walking, but also avoids spacer molecules, such as mercaptohexanol, which are generally required for electrode surface modification. The amino group marked on the DNA track can react with the silver nanoparticles through the silver-ammonia bond, so that the silver nanoparticles are positi...

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Abstract

The scheme of the invention relates to an electrochemical nucleic acid detection method based on DNA (deoxyribonucleic acid) walking and rolling circle amplification signal magnification. The method comprises the following steps: designing two DNA tetrahedrons, namely DNA walker and DNA track; immobilizing the DNA tetrahedrons on the surface of an electrode; adding a padlock probe into a sample tobe detected, inserting the electrode into the sample to be detected, and performing incubation; continuously adding DNA ligase and DNA polymerase into the sample to be detected, and performing a reaction; taking out the electrode, soaking the electrode into Pb<2+> at room temperature, performing a reaction, and finally performing incubation with silver nanoparticles; by taking the electrode as aworking electrode, and by using an electrochemical working station, recording a linear volt-ampere scanning curve by using a three-electrode system, detecting variation of volt-ampere current signals,and calculating the concentration of target nucleic acid. The detection method provided by the scheme of the invention is high in sensitivity, high in selectivity, simple and convenient to operate and low in detection cost.

Description

technical field [0001] The invention relates to the field of nucleic acid detection, in particular to an electrochemical nucleic acid detection method based on DNA walking (DNA walking) and rolling circle amplification signal amplification. Background technique [0002] Nucleic acid is the carrier of genetic information and the material basis of gene expression, and is the research focus of life science and clinical medicine. Nucleic acid detection technology has been applied to many fields such as basic research, drug screening, and environmental monitoring. Traditional nucleic acid detection methods include gel electrophoresis, polymerase chain reaction (PCR), etc., but there are still some shortcomings in practical applications. For example, dyes for gel electrophoresis staining are highly toxic, and PCR is prone to non-specific amplification and high false positives. In recent years, a series of nucleic acid sensing methods have been developed based on other technologi...

Claims

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Application Information

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IPC IPC(8): C12Q1/6851
CPCC12Q1/6851C12Q2531/125C12Q2565/607C12Q2521/501C12Q2563/137
Inventor 缪鹏柴华
Owner 天津国科医疗科技发展有限公司
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