Method for detecting miRNA-21 based on electrochemical alternating current impedance

A miRNA-21, AC impedance technology, applied in the field of electrochemical detection, can solve the problems of high sequence similarity, low abundance, short length, etc., and achieve the effect of strong anti-interference

Active Publication Date: 2019-06-07
宁波远志立方能源科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the short length, high sequence similarity and low abundance (as low as a few molecules per cell) of miRNAs make it difficult to develop fast, simple, sensitive and selective electrochemical methods for the detection of miRNAs

Method used

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  • Method for detecting miRNA-21 based on electrochemical alternating current impedance
  • Method for detecting miRNA-21 based on electrochemical alternating current impedance
  • Method for detecting miRNA-21 based on electrochemical alternating current impedance

Examples

Experimental program
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Effect test

Embodiment 1

[0030] A method for detecting miRNA-21 based on electrochemical impedance spectroscopy, comprising:

[0031] Step 1, grinding, polishing, washing and drying the gold electrode;

[0032] Step 2, gold electrode activation treatment:

[0033] a. Soak the surface of the gold electrode in step 1 in the miRNA-21 aptamer solution with a concentration of 2 μM for 1 min, then irradiate it with an electron beam with an intensity of 15 kGy for 3 min, and then soak it in the aptamer solution with a concentration of 10 μM at a temperature of 4°C. Glucose aqueous solution for 4 hours, then washed with PBS solution with pH 6.8 for 30 seconds, and dried with nitrogen;

[0034] b. Soak the surface of the gold electrode treated in step a with a PBS solution with a pH of 6.8, and place it in a magnetic field with a magnetic field strength of 1 T while soaking, take it out after 5 minutes, and rinse it with an ethanol solution of boric acid with a concentration of 5 μM for 15 minutes. Then rins...

Embodiment 2

[0048] The method for detecting miRNA-21 based on electrochemical AC impedance is the same as in Example 1, wherein step 1 is specifically:

[0049] The surface of the gold electrode was placed in NaHCO 3 In an ultrapure aqueous solution with a concentration of 5 μM and a concentration of KCl of 10 μM, soak for 40 minutes in a water bath at a temperature of 65 ° C, rinse with ultrapure water, and then place it in an ultrapure aqueous solution with a concentration of acetic acid of 5 μM and a concentration of silicic acid of 2 μM , soaked in a water bath at a temperature of 80°C for 60 minutes, rinsed with ultrapure water, polished with 0.05 μm polishing powder on a polishing cloth, and rinsed with ultrapure water.

Embodiment 3

[0051] The method for detecting miRNA-21 based on electrochemical impedance spectroscopy is the same as in Example 1, wherein the glucose aqueous solution in step a is prepared as follows: put ultrapure water at -10°C for 30 minutes, and add acetic acid to make the concentration of acetic acid 1 μM , placed in an environment at 4°C for 10 minutes, then placed in a freezer at -10°C for 10 minutes, added glucose, placed at a temperature of 4°C, and ultrasonicated at a frequency of 80KHz for 10 minutes.

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Abstract

The invention discloses a method for detecting miRNA-21 based on electrochemical alternating current impedance. The method comprises the following steps: firstly, burnishing a gold electrode, polishing and drying; secondly, soaking the surface of the gold electrode in miRNA-21 aptamer solution, and then sequentially performing electron beam irradiation, glucose aqueous solution soaking as well asflushing and drying; placing the gold electrode in a magnetic field to be soaked, flushing and drying; dropwise adding mercaptoethanol solution and performing reaction, flushing and drying, so that anactivated gold electrode is obtained; thirdly, preparing multiple activated gold electrodes, dropwise adding miRNA-21 solution with different concentrations, performing reaction, and then flushing and drying, so that multiple detection gold electrodes are obtained; fourthly, detecting an impedance value corresponding to the miRNA-21 solution with each concentration; fifthly, calculating to obtaina linear relationship between the concentrations and difference values of the impedance values; and sixthly, detecting an impedance value of to-be-detected sample solution, and calculating and obtaining the concentration of the miRNA-21 in the to-be-detected sample solution by adopting the linear relationship. The method disclosed by the invention has the beneficial effect of high detection sensitivity.

Description

technical field [0001] The invention relates to the field of electrochemical detection. More specifically, the present invention relates to a method for detecting miRNA-21 based on electrochemical AC impedance. Background technique [0002] MicroRNAs (miRNAs) are a class of small non-coding RNA molecules containing approximately 22 nucleotides that function in the transcriptional and post-transcriptional regulation of gene expression. According to some recent reports, certain types of miRNAs are closely related to human cancers, and miRNA-21 is one of them. Among these different miRNAs, microRNA-21 is considered as a potential biomarker because it is overexpressed in many cancer tissues, such as lung adenocarcinoma, glioblastoma, breast cancer, cervical cancer. Furthermore, by delivering inhibitors to regulate the expression of specific microRNAs, they could also serve as novel therapeutic targets for many diseases. Therefore, there is an urgent need for sensitive microRN...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/02
Inventor 肖琦黄珊李家文金晓宇
Owner 宁波远志立方能源科技有限公司
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