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Method and application of rapid detection of nucleic acid based on electrochemical potential pretreatment technology

An electrochemical and pretreatment technology, applied in the direction of biochemical equipment and methods, electrochemical variables of materials, microbial determination/inspection, etc.

Active Publication Date: 2021-05-25
HANGZHOU DIANZI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] One purpose of the present invention is to propose a method for directly and quickly detecting nucleic acid within a certain voltage range by using electrochemical potential pretreatment technology for the deficiencies of existing nucleic acid detection research

Method used

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  • Method and application of rapid detection of nucleic acid based on electrochemical potential pretreatment technology
  • Method and application of rapid detection of nucleic acid based on electrochemical potential pretreatment technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. DNA-DNA hybrid system

[0022] The artificially synthesized target DNA (5'-GGTGGGAAGGACCTGATGATAC-3') solution was dropped into 5pM positive matching probe (5'-GTATCATCAGGTCCTTCCCACC-3') solution and 5 pM mismatching probe (5'-GTATCATCAGG) solution respectively. A In the CCTTTCCCACC-3') solution, a positive hybridization solution and a mismatch hybridization solution are formed through the DNA-DNA hybridization reaction. The glassy carbon electrode was pretreated in the potential range of 1.8-2.7V, and then the simulated body fluid phosphate buffer solution (PBS buffer) with pH=7.4 was used as a blank comparison. The hybridization solution and the PBS blank solution were subjected to square wave voltammetry scanning, and the electrochemical sensing detection results were as follows figure 1 Shown: the square wave voltammetry curve of the positive hybridization solution has a shoulder peak in the range of 1.0-1.8V, and the square wave voltammetry curve of the...

Embodiment 2

[0023] Example 2. DNA-cDNA hybridization system

[0024] The cDNA (5'-TAACACTGTCTGGTAAAGATGG-3') solution of artificially synthesized prostate cancer characteristic marker miR-141 was dropped into 5pM positive matching probe (5'-CCATCTTTACCAGACAGTGTTA-3') solution and 5 pM mismatching probe solution respectively. Needle (5'- A In the CATCTTTACCAGACAGGTGTTA-3') solution, a positive hybridization solution and a mismatch hybridization solution are formed through the cDNA-DNA hybridization reaction. The glassy carbon electrode was pretreated in the potential range of 1.8-2.5V, and then the simulated body fluid phosphate buffer solution (PBS buffer) with pH=7.4 was used as a blank comparison. The hybridization solution and the PBS blank solution were scanned by square wave voltammetry, and the electrochemical sensing detection results were similar to those in the appendix. figure 1 , figure 2 shown.

Embodiment 3

[0025] Example 3. DNA-RNA hybrid system

[0026] The artificially synthesized target RNA (5'-GGUGGGAAGGACCUGAUGAUAC-3') solution was dropped into 5pM positive matching probe (5'-GTATCATCAGGTCCTTCCCACC-3') solution and 5 pM mismatching probe (5'-GT) solution respectively. T In TCATCAGGTCCTTCCCACC-3') solution, a positive hybridization solution and a mismatch hybridization solution are formed through DNA-RNA hybridization reaction. The glassy carbon electrode was pretreated in the potential range of 0-2.7V, and then the simulated body fluid phosphate buffer solution (PBS buffer) with pH=7.4 was used as a blank comparison. The hybridization solution and the PBS blank solution were scanned by square wave voltammetry, and the electrochemical sensing detection results were similar to those in the appendix. figure 1 , figure 2 shown.

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Abstract

The invention discloses a method and application for rapidly detecting nucleic acid based on electrochemical potential pretreatment technology. In the method, the PBS buffer is used as the electrolyte, the probe and the target nucleic acid fragment are added into the PBS buffer, and a nucleic acid hybridization solution is formed according to the principle of complementary base pairing. The sensing electrode is subjected to electrochemical pretreatment in a potential range greater than 1.8V, and then SWV is used to selectively detect target nucleic acids. The nucleic acid detection method of the electrochemical pretreatment technology is applied in the fields of disease characteristic miRNA-related electrochemical biosensing detection, medical diagnosis, and pathogen detection in food and environment. When this detection method is applied to nucleic acid, it has the following characteristics: high selectivity, which can detect single base mismatches for nucleic acid molecules; high sensitivity, with a detection sensitivity of 5×10 ‑18 M; the test results can be completely repeated; the operation process is economical and simple, there is no chain reaction, no amplification, no fluorescent indicator.

Description

technical field [0001] The invention belongs to the technical field of microelectronic biosensing, and relates to a nucleic acid fragment detection method of electrochemical pretreatment technology, in particular to a direct detection method of miRNA using electrochemical potential pretreatment technology and the application of miRNA related electrochemical sensing detection . Background technique [0002] miRNA is a kind of non-coding small molecule single-stranded RNA, which widely exists in animals, plants and protists, and participates in regulating various biological processes of biological systems by way of gene expression. Therefore, the application of miRNA detection has aroused wide interest in application fields including medicine, food, forensics and environment, and is currently a hot spot in the field of sensor detection applications. In particular, miRNAs play a key role in pathological processes such as inflammation and carcinogenesis, and small RNAs such as ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6825G01N27/48
CPCC12Q1/6825G01N27/48
Inventor 崔静洁黄震徐平李宏
Owner HANGZHOU DIANZI UNIV
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