Methylation electrochemical immunoassay method, electrode and electrochemical sensor

Abstract

The invention belongs to the technical field of immunoassay, and discloses a methylation electrochemical immunoassay method, an electrode and an electrochemical sensor. The methylation electrochemical immunoassay method comprises the steps of: preparing a nanogold modified gold electrode, and preparing the electrochemical sensor; and immobilizing the anti-5-mC antibody and the enzyme-labeled secondary antibody, and performing electrochemical determination. The preparation method of the nanogold-modified gold electrode comprises the following steps: polishing a gold electrode with the diameter of 2mm into a mirror surface by using 0.05 [mu]m Al2O3 powder, and then carrying out ultrasonic cleaning for 5 minutes in ultrapure water, absolute ethyl alcohol and ultrapure water respectively; after drying at room temperature, putting the electrode into a prepared Piranha solution to activate for more than 15 minutes, then thoroughly cleaning with ultrapure water, and blow-drying with nitrogen; and immersing the treated gold electrode in a HAuCl4. 3H2O solution, and finally, fuly washing and cleaning the gold electrode with ultrapure water, and preparing the modified gold electrode. Along with the increase of the number of DNA methylation sites, the peak current of the DPV is increased.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and in particular relates to a methylation electrochemical immunoassay method, an electrode and an electrochemical sensor. Background technique [0002] At present, DNA methylation has always been the frontier and hot spot in the field of epigenetic research. Because it plays an important role in the regulation of gene expression in cell differentiation, embryonic development, genetic diseases and tumorigenesis, DNA methylation analysis has become a New tools for early warning of epigenetic disorders and tumors. There are certain deficiencies in the traditional DNA methylation detection technology, such as sequencing depth, conversion efficiency, and huge data analysis in bisulfite sequencing, and the design of primers for methylation restriction enzyme digestion analysis is limited by the enzyme recognition sequence, etc. Operations such as liquid chromatography and mass spectrometry ar...

AI Technical Summary

Problems solved by technology

There are certain deficiencies in the traditional DNA methylation detection technology, such as sequencing depth, conversion efficiency, and huge data analysis in bisulfite sequencing, and the design of primers for methylation restriction enzyme digestion analysis is limited by the enzyme recognition sequence, etc. Liquid chromatography, mass spectrometry and other operations are cumbersome and require huge equipment

Electrochemical sensing analysis and detection technology overcomes the above shortcomings, and has many advantages such as low cost, simple operation, fast response, high sensitivity and easy miniaturization of equipment. The application of electrochemical sensors at home and abroad to detect DNA methylation is mainly through the detection of DNA methylation The activity of base transferase indirectly reflects the level of DNA methylation, and the sensitivity and specificity still need to be improved

[0003] Through the above analysis, the existing problems and defects of the prior art are: the application of electrochemical sensors to detect DNA methylation at home and abroad mainly reflects the low level of DNA methylation indirectly through the detection of DNA methyltransferase activity, and the sensitivity is poor. poor specificity

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