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Method and application for rapidly detecting nucleic acid based on electrochemical electric potential pretreatment technology

A technology of electrochemistry and pretreatment, which is applied in the fields of biochemical equipment and methods, electrochemical variables of materials, determination/inspection of microorganisms, etc., to achieve the effect of simple and economical operation.

Active Publication Date: 2018-05-04
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 for rapidly detecting nucleic acid based on electrochemical electric potential pretreatment technology
  • Method and application for rapidly detecting nucleic acid based on electrochemical electric potential pretreatment technology

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Embodiment 1. DNA-DNA hybrid system

[0022] The artificially synthesized target DNA (5'-GGTGGGAAGGACCTGATGATAC-3') solution was dropped into equal volumes of 5pM positive match probe (5'-GTATCATCAGGTCCTTCCCCACC-3') solution and 5pM mismatch probe (5'-GTATCATCAGGACCTTCCCACC-3 ') solution, a positive hybridization solution and a mismatched hybridization solution are formed through the DNA-DNA hybridization reaction. The glassy carbon electrode is 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 is used as a blank comparison. The hybridization solution and the PBS blank solution were subjected to square wave voltammetry scanning, and the electrochemical sensing results were as attached figure 1 Shown: shoulder peak occurs in the square wave voltammetry curve of normal matching hybridization solution in the range of 1.0-1.8V, see figure 1 a, the square wave voltammetry curves of the mismat...

Embodiment 2

[0023] Example 2. DNA-cDNA hybrid 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 5pM mismatching probe respectively in equal volumes. In the needle (5'-ACATCTTTACCAGACAGTGTTA-3') solution, a positive hybridization solution and a mismatched hybridization solution are formed through cDNA-DNA hybridization reaction. The glassy carbon electrode is 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 is used as a blank comparison. The hybridization solution and the PBS blank solution were scanned by square wave voltammetry, and the electrochemical sensing results were similar to those in the attached figure 1 , figure 2 shown.

Embodiment 3

[0025] Embodiment 3.DNA-RNA hybrid system

[0026] The synthetic target RNA (5'-GGUGGGAAGGACCUGAUGAUAC-3') solution was dropped into 5pM positive matching probe (5'-GTATCATCAGGTCCTTCCCCACC-3') solution and 5pM mismatching probe (5'-GTTTCATCAGGTCCTTCCCACC-3 ') solution, a positive hybridization solution and a mismatched hybridization solution are formed through the DNA-RNA hybridization reaction. The glassy carbon electrode is pretreated in the potential range of 0-2.7V, and then the simulated body fluid phosphate buffer solution (PBS buffer) with pH=7.4 is used as a blank comparison. In the potential range of less than 1.8V, the positive hybridization solution and mismatch The hybridization solution and the PBS blank solution were scanned by square wave voltammetry, and the electrochemical sensing results were similar to those in the attached figure 1 , figure 2 shown.

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Abstract

The invention discloses a method and application for rapidly detecting nucleic acid based on an electrochemical electric potential pretreatment technology. The method comprises the following steps: taking a PBS buffer solution as electrolyte, adding a probe and target nucleic acid fragments into the PBS buffer solution, and forming a nucleic acid hybrid solution according to a base complementationpairing rule. A sensor electrode is subjected to electrochemical pretreatment in an electric potential range of more than 1.8V, and then target nucleic acids are selectively detected by using SWV. The invention further discloses application of a nucleic acid detection method of the electrochemical pretreatment technology in disease characteristic miRNA related electrochemical bio-sensing detection and medical diagnosis and in the detection field of pathogens in foods, environments and the like. When applied to nucleic acids, the detection method has the following characteristics that the detection method has high selectivity and can realize single-base mismatch detection on nucleic acid molecules; the method has high sensitivity, and the detection sensitivity reaches 5*10<-18>M; the detection result can be completely repeated; and the operation process is economical and simple, any chain reaction does not exist, amplification is not needed, and any fluorescence indicator is not needed.

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 miRNA detection method of 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 the regulation of various biological processes in biological systems through gene expression. Therefore, the application of miRNA detection has aroused widespread interest in application fields including medicine, food, forensic medicine and the environment, and is currently a hot spot in the field of sensor detection application research. In particular, miRNAs play a key role in pathological processes such as inflammation and carcinogenesis...

Claims

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Application Information

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Patent Type & Authority Applications(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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