CN112014446B - Sensor for detecting gene methylation level and preparation and detection methods thereof - Google Patents
Sensor for detecting gene methylation level and preparation and detection methods thereof Download PDFInfo
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Abstract
The invention provides an electrochemical sensor for detecting gene methylation level based on a terminal labeling strategy, and a preparation method and a detection method thereof. The electrochemical sensor takes a glassy carbon electrode as a substrate electrode, takes gold nanoparticles as an electrode modification material, and is modified to obtain a modified electrode. Modifying a sulfydryl at the 5' end of a gene to be detected by utilizing a terminal labeling strategy, directly fixing the gene to be detected on the surface of a modified electrode through covalent bond interaction, combining a specific antibody to a methylation site of the gene to be detected by utilizing the interaction of the antibody and an antigen, combining a secondary antibody labeled with HRP to the specific antibody through the interaction of a primary antibody and a secondary antibody, catalyzing the redox reaction of hydrogen peroxide and hydroquinone by the HRP to generate a signal, and detecting the methylation level of the gene to be detected. The electrochemical sensor is not influenced by the base sequence of the gene to be detected, has wide detection range and high repeatability, and has great potential in clinical research and practical application.
Description
Technical Field
The invention belongs to the field of electrochemical biosensors, and particularly relates to an electrochemical sensor for detecting gene methylation level based on a terminal labeling strategy, and a preparation method and a detection method thereof.
Background
Gene methylation is the most important mechanism in epigenetics, and refers to the chemical modification process of introducing a methyl group into a specific base on a DNA or RNA sequence under the catalysis of methyltransferase. Gene methylation plays an important role in physiological and pathological regulation, and abnormal gene methylation is often associated with a variety of genetic diseases and tumor production. Therefore, effective detection of the methylation level of a gene is of great significance for diagnosis of early stage tumors, research of genetic information control mechanisms, and the like.
Electrochemical sensors have attracted much attention due to their advantages of high sensitivity, high selectivity, low cost, short time consumption, easy operation, etc. At present, most of electrochemical sensors adopt DNA probes synthesized by companies to capture and identify target genes. The electrochemical sensor prepared in this way has a disadvantage that it is difficult to apply to an actual genome for detection. Since the DNA probes synthesized by the company are all fixed base sequences, it is determined that the probe can capture only a gene strand complementary to its base sequence. However, in the actual genome, the base sequences of methylated genes are diverse, which limits the application of electrochemical sensors in the detection of gene methylation. Therefore, there is a need for an electrochemical sensor that does not use a DNA probe and is not affected by the nucleotide sequence of a gene.
Disclosure of Invention
The invention aims to provide an electrochemical sensor for detecting the gene methylation level based on a terminal marker strategy, a preparation method and a detection method thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides an electrochemical sensor for detecting the methylation level of a gene based on a terminal labeling strategy, which is characterized in that a glassy carbon electrode is used as a substrate electrode, gold nanoparticles are used as electrode modification materials and are modified to obtain a modified electrode, the 5' end of the gene to be detected is modified with a sulfydryl group by using the terminal labeling strategy, the gene to be detected is directly fixed on the surface of the modified electrode through covalent bond interaction, then a specific antibody for identifying the methylation site in the gene to be detected is combined to the methylation site of the gene to be detected by using the interaction of the antibody and an antigen, and then a secondary antibody marked with HRP is combined to the specific antibody through the interaction of a primary antibody and a secondary antibody, the HRP catalyzes the redox reaction of hydrogen peroxide and hydroquinone to generate a signal, so that the methylation level of the gene to be detected is detected.
The second aspect of the present invention provides a method for preparing an electrochemical sensor for detecting the methylation level of a gene based on a terminal labeling strategy, comprising the following steps:
step 1: modifying sulfydryl at the 5' end of the methylated gene to be detected by adopting a terminal marking strategy;
step 2: polishing the glassy carbon electrode by using 0.05 mu m of aluminum oxide polishing powder, respectively ultrasonically cleaning by using ethanol and water, and then drying by using nitrogen; immersing the treated glassy carbon electrode into a chloroauric acid solution, and electrodepositing gold nanoparticles on the surface of the glassy carbon electrode by adopting a potentiostatic method; washing with ultrapure water and drying in the air to obtain a gold nanoparticle modified glassy carbon electrode;
and step 3: dropwise coating the methylation gene to be detected, which is subjected to sulfhydrylation modification and is obtained in the step 1, on the surface of the electrode, which is modified by the gold nanoparticles and obtained in the step 2, and incubating overnight; after the PBS solution is cleaned, dripping MCH solution, sealing for 1h, then dripping BSA solution and incubating for half an hour; after washing, dropwise adding an antibody solution, incubating for 1h at 37 ℃, and dropwise adding a secondary antibody solution marked with HRP, incubating for 1h at 37 ℃; and washing and airing the PBS to obtain the electrochemical sensor.
Further, the step 1 specifically includes: the method comprises the steps of mixing T4 polynucleotide kinase, adenosine 5' - [ gamma-thio ]]Triphosphates, the methylated gene to be assayed, dissolved in 70mM Tris-HCl buffer (pH 7.6, containing 10mM MgCl)2And 5mM DTT), overnight.
Further, in the step 2, the concentration of the chloroauric acid solution is 3mM, and the constant potential is-0.2V.
Further, in the step 3, the concentration of the methylated gene to be detected is 10–12～10-7M。
In a third aspect of the invention, a method for detecting an electrochemical sensor based on a terminal labeling strategy for detecting the methylation level of a gene is provided, wherein the electrochemical sensor is placed in a phosphate buffer solution containing hydrogen peroxide and hydroquinone and is detected by adopting differential pulse voltammetry.
Compared with the prior art, the invention has the following advantages:
according to the electrochemical sensor for detecting the gene methylation level based on the terminal labeling strategy, the gene to be detected is modified with the sulfhydryl group at the 5' end through a simple and mild reaction, and then the gene to be detected can be directly fixed on the surface of the glassy carbon electrode modified with gold nanoparticles through Au-S covalent interaction, so that the capture process of a DNA probe is not needed, and the detection of the gene methylation level is not influenced by a base sequence. In addition, the antibody is combined with the specificity recognition of the methylation site in the gene to be detected, the specific detection of the methylation of a specific base can be realized, and the selectivity of the sensor is greatly improved. The application of the gold nanoparticles and the HRP in the preparation of the sensor not only amplifies signals, but also improves the sensitivity of the sensor. The electrochemical sensor has wide detection range and high repeatability, and has great potential in clinical research and practical application.
Drawings
FIG. 1 is a schematic diagram of an electrochemical sensor for detecting the methylation level of a gene based on a terminal labeling strategy according to the present invention;
FIG. 2 is a diagram showing the response of electrochemical signals corresponding to genes to be detected obtained by different processing methods according to the present invention;
in the figure, a curve a is a blank, namely the glassy carbon electrode modified by the gold nanoparticles; curve b is the gene to be tested which has not been processed by the end-labeling strategy; curve c is the gene to be detected after the treatment of the terminal marking strategy;
FIG. 3 is a diagram showing the response of electrochemical signals of the electrochemical sensor according to the present invention to genes to be detected in different states;
in the figure, a curve a is a methylated gene to be detected which is processed by a terminal marking strategy; curve b is the unmethylated gene to be detected which is treated by the end-labeling strategy; curve c is the methylated gene to be detected which has not been subjected to the end-labeling strategy; curve d is blank;
FIG. 4 is an electrochemical signal response diagram corresponding to methylated genes to be detected at different concentrations and a linear relationship between signal response and concentration according to the present invention;
in the figure, the concentrations represented by a-f are, in order: 10-12M，10-11M，10-10M，10-9M，10-8M， 10-7M。
Detailed Description
The features and advantages of the present invention will be further understood from the following detailed description taken in conjunction with the accompanying drawings. The examples provided are merely illustrative of the method of the present invention and do not limit the remainder of the disclosure in any way.
Example 1 treatment of methylated genes to be tested with end-labeling strategy
1. Synthesizing a sulfydryl modified methylation gene to be detected: the method comprises the steps of mixing T4 polynucleotide kinase, adenosine 5' - [ gamma-thio ]]Triphosphates, the methylated gene to be assayed, dissolved in 70mM Tris-HCl buffer (pH 7.6, containing 10mM MgCl)2And 5mM DTT), reacting overnight;
2. extracting, purifying and synthesizing the gene to be detected: water and phenol were added: chloroform: isoamyl alcohol (25:24:1), mixing and centrifuging, and extracting supernatant; adding 3M sodium acetate buffer solution (pH 5.2), 20mg/mL glycogen and absolute ethyl alcohol, mixing, and freezing and incubating for 2 h; and centrifuging, collecting the precipitate, washing with 70% ethanol, air-drying, and adding primary water to obtain the purified methylation gene to be detected, which is modified by sulfhydrylation.
EXAMPLE 2 preparation and detection of electrochemical sensor
Taking N6-methyladenine (6mA) modified DNA as an example, 6mA is another important methylation modification in DNA which has attracted much attention in recent years. FIG. 1 is a schematic diagram showing the preparation of an electrochemical sensor based on the end-labeling strategy and its detection of the level of 6mA modification in DNA. Firstly, polishing and cleaning a glassy carbon electrode, electrodepositing gold nanoparticles on the surface of the glassy carbon electrode at a constant potential of-0.2V, dripping DNA-6mA treated by a terminal marking strategy on the surface of a modified electrode, and incubating overnight; after the PBS solution is cleaned, dripping MCH solution, sealing for 1h, then dripping BSA solution and incubating for half an hour; after washing, dropwise adding 6mA antibody solution, incubating for 1h at 37 ℃, and dropwise adding secondary antibody solution marked with HRP, incubating for 1h at 37 ℃; and washing with PBS and drying to obtain the electrochemical sensor. The method comprises the following specific steps:
(1) polishing the glassy carbon electrode by using 0.05 mu m of aluminum oxide polishing powder, respectively ultrasonically cleaning by using ethanol and water, and then drying by using nitrogen; immersing the treated glassy carbon electrode into a chloroauric acid solution, and electrodepositing gold nanoparticles on the surface of the glassy carbon electrode by adopting a potentiostatic method; washing with ultrapure water and drying in the air to obtain a gold nanoparticle modified glassy carbon electrode (modified electrode);
(2) dripping the obtained methylation gene to be detected, which is subjected to sulfhydrylation modification, on the surface of the electrode modified by the gold nanoparticles obtained in the step (1), and incubating overnight; after the PBS solution is cleaned, dripping MCH solution, sealing for 1h, then dripping BSA solution and incubating for half an hour; after washing, dropwise adding an antibody solution, incubating for 1h at 37 ℃, and dropwise adding a secondary antibody solution marked with HRP, incubating for 1h at 37 ℃; and washing and airing the PBS to obtain the electrochemical sensor.
The electrochemical sensor is placed in a phosphate buffer solution containing hydrogen peroxide and hydroquinone, and the detection is carried out by adopting differential pulse voltammetry.
Wherein, in the step (1), the concentration of the chloroauric acid solution is 3mM, and the constant potential is-0.2V; in the step (2), the concentration of the methylated gene to be detected is 10–12～10-7M。
In the method, T4 polynucleotide kinase used in the end labeling strategy can catalyze the transfer of the gamma-position phosphorothioate group of adenosine 5' - [ gamma-thio ] triphosphate to the 5' -hydroxyl end of the gene to be detected (double-stranded or single-stranded DNA or RNA), so that the 5' end of the gene to be detected is provided with a sulfhydryl group; through the covalent interaction of Au-S, the gene to be detected can be directly fixed on the surface of the glassy carbon electrode modified by the gold nanoparticles; the antibody can specifically identify and combine with a methylation site in a gene to be detected; then, the secondary antibody marked with HRP can be specifically combined with the antibody; HRP is horseradish peroxidase, can catalyze the redox reaction of hydrogen peroxide and hydroquinone, produce the electron transfer, this process can be monitored by the electrochemical workstation, and turn into the electric signal that can be detected quantitatively; the more methylation sites in the gene to be detected, the larger the electric signal, so that the electrochemical detection of the methylation level of the gene can be realized.
Example 3 comparison of electrochemical Signal response corresponding to genes to be tested obtained by different treatment methods
In order to verify that the terminal labeling strategy can modify a to-be-detected gene with a sulfhydryl group and fix the to-be-detected gene on the surface of a modified electrode, respectively dripping the to-be-detected gene which is not processed by the terminal labeling strategy and the processed to-be-detected gene on a glassy carbon electrode modified by gold nanoparticles, and incubating overnight; and then, respectively placing the modified electrodes into potassium ferricyanide solution for cyclic voltammetry measurement, wherein the result is shown in fig. 2, and the curve c signal response corresponding to the processed gene to be detected is minimum, because the gene to be detected can be greatly fixed on the surface of the modified electrodes after passing through the sulfhydryl group on the processing belt, thereby blocking electron transfer and causing minimum current response. The difference between the curve a corresponding to the untreated gene to be detected and the curve b corresponding to the blank is almost the same, which indicates that the adopted terminal marking strategy can be effectively applied to the sulfhydrylation modification of the gene to be detected.
Example 4 comparison of electrochemical Signal response of electrochemical sensor to Gene to be measured in different states
The treated methylated gene, the treated unmethylated gene, and the untreated methylated gene were detected by the prepared sensors, respectively, and the detection methods were prepared in the same manner as in example 2. As a result, as shown in FIG. 3, the signal response of the curve a corresponding to the processed methylated gene is the largest, and the signal response of the gene to be detected in other states is not greatly different from that of the blank, which indicates that the constructed electrochemical sensor has high selectivity for detecting the methylated gene, and the sensor can be effectively applied to detecting the methylated gene.
Example 5 high sensitivity detection of methylated genes
The prepared sensor is used for detecting DNA containing 6mA modification at different concentrations, and the preparation and detection method is the same as that of example 2. As can be seen from FIG. 4, the response value of the electric signal is increasing with the increase of the concentration of DNA-6mA, and is 10-12M to 10-7M exhibits a good linear relationship.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and it will be apparent to those skilled in the art that several modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should be construed as the protection scope of the present invention.
Claims (6)
1. An electrochemical sensor for detecting the methylation level of a gene based on a terminal labeling strategy, which is characterized in that: the electrochemical sensor takes a glassy carbon electrode as a substrate electrode, gold nanoparticles as an electrode modification material to obtain a modified electrode through modification, a terminal labeling strategy is utilized to modify a 5' end of a gene to be detected with a sulfydryl group, the gene to be detected is directly fixed on the surface of the modified electrode through covalent bond interaction, a specific antibody for identifying a methylation site in the gene to be detected is combined to the methylation site of the gene to be detected through the interaction of an antibody and an antigen, a secondary antibody labeled with HRP is combined to the specific antibody through the interaction of a primary antibody and a secondary antibody, the HRP catalyzes the redox reaction of hydrogen peroxide and hydroquinone to generate a signal, and the methylation level of the gene to be detected is detected.
2. A method for preparing the electrochemical sensor for detecting the methylation level of a gene based on the terminal labeling strategy according to claim 1, wherein the electrochemical sensor comprises: the method comprises the following steps:
step 1: modifying sulfydryl at the 5' end of the methylated gene to be detected by adopting a terminal marking strategy;
step 2: polishing the glassy carbon electrode by using 0.05 mu m of aluminum oxide polishing powder, respectively ultrasonically cleaning by using ethanol and water, and then drying by using nitrogen; immersing the treated glassy carbon electrode into a chloroauric acid solution, and electrodepositing gold nanoparticles on the surface of the glassy carbon electrode by adopting a potentiostatic method; washing with ultrapure water and drying in the air to obtain a gold nanoparticle modified glassy carbon electrode;
and step 3: dropwise coating the methylation gene to be detected, which is subjected to sulfhydrylation modification and is obtained in the step 1, on the surface of the electrode, which is modified by the gold nanoparticles and obtained in the step 2, and incubating overnight; after the PBS solution is cleaned, dripping MCH solution, sealing for 1h, then dripping BSA solution and incubating for half an hour; after washing, dropwise adding an antibody solution, incubating for 1h at 37 ℃, and dropwise adding a secondary antibody solution marked with HRP, incubating for 1h at 37 ℃; and washing and airing the PBS to obtain the electrochemical sensor.
3. The method for preparing an electrochemical sensor for detecting the methylation level of a gene according to claim 2, wherein the electrochemical sensor comprises: the above-mentionedThe step 1 specifically comprises the following steps: mixing T4 polynucleotide kinase, adenosine 5' - [ gamma-thio ]]Dissolving triphosphate and methylated gene to be detected in 70mM Tris-HCl buffer solution for overnight reaction; wherein the Tris-HCl buffer has a pH of 7.6 and contains 10mM MgCl2And 5mM DTT.
4. The method for preparing an electrochemical sensor for detecting the methylation level of a gene according to claim 2, wherein the electrochemical sensor comprises: in the step 2, the concentration of the chloroauric acid solution is 3mM, and the constant potential is-0.2V.
5. The method for preparing an electrochemical sensor for detecting the methylation level of a gene according to claim 2, wherein the electrochemical sensor comprises: in the step 3, the concentration of the methylation gene to be detected is 10–12～10-7M。
6. A method for detecting the methylation level of a gene by using an electrochemical sensor based on a terminal labeling strategy, which is prepared by the method of claim 2, wherein the electrochemical sensor comprises: and (3) placing the electrochemical sensor in a phosphate buffer solution containing hydrogen peroxide and hydroquinone, and detecting by adopting differential pulse voltammetry.
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| CN101769892A (en) * | 2010-01-26 | 2010-07-07 | 湖南农业大学 | Electrochemistry immuno-sensing method for detecting 2,4-dichlorphenoxyacetic acid in agricultural product |
| CN105116149A (en) * | 2015-06-04 | 2015-12-02 | 塔里木大学 | Electrochemical immunosensor for detecting pesticide azacyclotin and preparation method thereof |
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| CN105116149A (en) * | 2015-06-04 | 2015-12-02 | 塔里木大学 | Electrochemical immunosensor for detecting pesticide azacyclotin and preparation method thereof |
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| A bisulfite treatment and PCR-free global DNA methylation detection method using electrochemical enzymatic signal engagement;Ripon Bhattacharjee等;《Biosensors and Bioelectronic》;20181012;第126卷;第102页摘要，第103页左栏最后一段和Experimental * |
| 基于5-甲基胞嘧啶抗体的DNA甲基化电化学检测;粟莎莎等;《贵州医科大学学报》;20191130;第44卷(第11期);第1262页摘要，第1263-1266页材料与方法，结果 * |
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