A Method for Detection of miRNA-21 Based on Electrochemical AC Impedance

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

Active Publication Date: 2021-10-15
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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  • A Method for Detection of miRNA-21 Based on Electrochemical AC Impedance
  • A Method for Detection of miRNA-21 Based on Electrochemical AC Impedance
  • A Method for Detection of miRNA-21 Based on Electrochemical AC Impedance

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Experimental program
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Effect test

Embodiment 1

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

[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 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 glucose with a concentration of 10 μM at a temperature of 4 °C In aqueous solution for 4h, then rinsed with PBS solution of pH 6.8 for 30s, and dried with nitrogen;

[0034] b. Soak the gold electrode surface 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 rinse with PBS solution with a pH of 6.8 for 30s and dry w...

Embodiment 2

[0047] The method for detecting miRNA-21 based on electrochemical AC impedance is the same as that in Embodiment 1, wherein step 1 is specifically:

[0048] The gold electrode surface was placed in NaHCO 3 In an ultrapure aqueous solution with a concentration of 5 μM and a concentration of KCl of 10 μM, immersed in a water bath at a temperature of 65 °C for 40 minutes, rinsed with ultrapure water, and then placed 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 0.05μm polishing powder on a polishing cloth, and rinsed with ultrapure water.

Embodiment 3

[0050] The method for detecting miRNA-21 based on electrochemical AC impedance is the same as that in Example 1, wherein, the configuration method of the aqueous glucose solution in step a is: placing ultrapure water at -10°C for 30 minutes, adding acetic acid to make the acetic acid concentration 1 μM , placed in a 4°C environment for 10min, then placed in a freezer at -10°C for 10min, added glucose, placed at a temperature of 4°C, the ultrasonic frequency was 80KHz, and the ultrasonic treatment was performed for 10min.

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Abstract

The invention discloses a method for detecting miRNA-21 based on electrochemical AC impedance. Perform electron beam irradiation, soak in glucose aqueous solution, rinse and dry; place in a magnetic field for soaking, rinse and dry; dropwise add mercaptoethanol solution for reaction, rinse and dry to obtain activated gold electrodes; step 3, prepare multiple activated gold electrodes, drip Add different concentrations of miRNA-21 solutions, rinse and dry after the reaction, and obtain multiple detection gold electrodes; step 4, detect the impedance value corresponding to each concentration of miRNA-21 solution; step 5, calculate the difference between the impedance value and The linear relationship of the concentration; step 6, detecting the impedance value of the sample solution to be tested, and calculating the concentration of miRNA-21 in the sample solution to be tested by using the linear relationship. The invention has the beneficial effect of high detection sensitivity.

Description

technical field [0001] The present 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 about 22 nucleotides whose function is the transcriptional and post-transcriptional regulation of gene expression. According to some recent reports, certain kinds of miRNAs are closely related to human cancer, 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. In addition, by delivering inhibitors to modulate specific microRNA expression, they can also be novel therapeutic targets for many diseases. Therefore, there is an urgent need for sensitive microRNA quantification...

Claims

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Application Information

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Patent Type & Authority Patents(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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