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Copper-hexahydroxy triphenylene-based miRNA-21 (micro ribonucleic acid-21) electrochemical sensor and detecting method thereof

A technology of hydroxytriphenylene and miRNA-21, which is applied in the field of electrochemical detection, can solve the problems that signal changes are greatly affected by the detection environment, the signal molecule labeling process is complicated, and the signal molecule and substrate electrode types are single.

Active Publication Date: 2019-03-12
MINNAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the probe-immobilized nucleic acid sensor, the probe-free immobilized nucleic acid sensor has faster hybridization reaction speed and higher hybridization efficiency, but at the same time, there are complex signal molecule labeling processes, and signal changes caused by kinetic diffusion differences are affected. Disadvantages such as large impact on the detection environment, single types of signal molecules and substrate electrodes, etc.

Method used

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  • Copper-hexahydroxy triphenylene-based miRNA-21 (micro ribonucleic acid-21) electrochemical sensor and detecting method thereof
  • Copper-hexahydroxy triphenylene-based miRNA-21 (micro ribonucleic acid-21) electrochemical sensor and detecting method thereof
  • Copper-hexahydroxy triphenylene-based miRNA-21 (micro ribonucleic acid-21) electrochemical sensor and detecting method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0035] The detection method of the miRNA-21 electrochemical sensor based on copper-hexahydroxytriphenylene, the specific steps are as follows:

[0036] Step 1: Place the pretreated glassy carbon electrode on Cu(NO 3 ) 2 and N,N-methylpyrrolidone (NMP) mixed solution for i-t scanning electrodeposition of Cu simple film, Cu(NO 3 ) 2 The concentration of N,N-methylpyrrolidone (NMP) is 50mmol / L, and the concentration of N,N-methylpyrrolidone (NMP) is 50mmol / L; then the electrode is scanned by cyclic voltammetry in the mixture of CAT and NMP, the concentration of CAT is 50mmol / L, and the concentration of NMP The concentration is 50mmol / L, then washes with twice distilled water, obtains Cu-CAT modified sensing electrode, is recorded as: Cu-CAT / GCE;

[0037] Step 2: Mix the DNA complementary to miRNA-21 and miRNA-21. After mixing, the DNA concentration is 10nmol / L, and the miRNA-21 concentration is 10pmol / L. The mixture is incubated at 15-50°C for 60min to form DNA-RNA After comp...

Embodiment 2

[0048] Preparation of Modified Electrode 1: Put the Cu-CAT / GCE prepared in Step 1 of Example 1 in a solution of 10nmol / L single-stranded probe sequence (DNA: 5'-CAG TCT GAT AAG CTA-3') at 37°C After soaking at constant temperature for 50 minutes, rinse with double distilled water to obtain modified electrode 1;

[0049] Preparation of modified electrode 2: Mix DNA and miRNA-21 that are complementary to miRNA-21. After mixing, the DNA concentration is 10nmol / L, and the miRNA-21 concentration is 10pmol / L. React at a constant temperature of 37°C for 60min to form DNA-RNA To compound the chain, immerse the Cu-CAT / GCE prepared in step 1 in Example 1 into the DNA-RNA complex chain reaction solution at 37°C for 50 minutes, and then rinse with double distilled water to obtain the modified electrode 2;

[0050] The copper-hexahydroxytriphenylene-based miRNA-21 electrode obtained in step 2 of Example 1 was set as modified electrode 3;

[0051] Such as image 3 As shown, the differenti...

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Abstract

Disclosed are a copper-hexahydroxy triphenylene-based miRNA-21 electrochemical sensor and a detecting method thereof. The copper-hexahydroxy triphenylene-based miRNA-21 electrochemical sensor is in athree-electrode system, wherein an auxiliary electrode is a platinum electrode, a reference electrode is an Ag / AgCl electrode, and a working electrode is a copper-hexahydroxy triphenylene-based miRNA-21 eletrode. The copper-hexahydroxy triphenylene-based miRNA-21 electrochemical sensor is characterized in that the copper-hexahydroxy triphenylene-based miRNA-21 electrode is modified with a Cu-CAT (catalase) membrane, and the Cu-CAT membrane absorbs nucleotide products produced by shearing DNA / miRNA-21 (deoxyribonucleic acid / miRNA-21) with ExoIII (exonuclease III). Through the specific hydrophobic pi-pi stack effects between Cu-CAT and oligonucleotide products sheared by ExoIII, the copper-hexahydroxy triphenylene-based miRNA-21 electrochemical sensor can achieve high-sensitivity, rapid andprobe fixing-free sensing analysis on HCC (hepatocellular carcinoma) biomarkers.

Description

technical field [0001] The invention relates to the field of electrochemical detection, in particular to a copper-hexahydroxytriphenylene-based miRNA-21 electrochemical sensor and a detection method. Background technique [0002] MicroRNA (miRNA) is a kind of extracellular nucleic acid in a cell-free state, widely present in body fluids such as peripheral blood, cerebrospinal fluid and urine. A large number of studies in recent years have shown that miRNA-21 is closely related to the occurrence, metastasis and drug resistance of hepatocellular carcinoma (HCC), and can be used as a specific biomarker of HCC. Therefore, it is of great significance to establish a rapid, accurate and sensitive miRNA-21 detection technology for the early warning, treatment and molecular mechanism research of HCC. [0003] Electrochemical nucleic acid sensors have the advantages of easy miniaturization, high sensitivity, and simple instruments, and are often used in the detection of miRNA. The c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3277G01N27/48
Inventor 高凤宋娟汪庆祥
Owner MINNAN NORMAL UNIV
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