CN113588752A - Preparation method and application of electrochemiluminescence aptamer sensor - Google Patents
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Abstract
The invention provides a preparation method of an electrochemiluminescence aptamer sensor for inhibiting ruthenium-based metal organic framework preparation based on a bismuth sulfide nanorod and detection of vomitoxin by the electrochemiluminescence aptamer sensor, wherein the electrochemiluminescence aptamer sensor is used as an electrochemiluminescence probe and further functionalizes the vomitoxin aptamer based on large specific surface area and high luminous efficiency of Ru-MOFs nano materials, and meanwhile, Bi is used2S3Labeling the aptamer complementary strand, the aptamer binding to the complementary strand such that Bi2S3Acts on the Ru-MOFs nano material to inhibit the ECL signal thereof, and realizes the high-sensitivity and high-stability detection of the vomitoxin by utilizing the specific recognition of the aptamer to the vomitoxin.
Description
Technical Field
The invention belongs to the technical field of functional materials and biosensing detection, and particularly relates to a preparation method and application of an electrochemiluminescence aptamer sensor for inhibiting a ruthenium-based metal organic framework based on a bismuth sulfide nanorod.
Background
Deoxynivalenol (DON), also known as vomitoxin, is mainly produced by fusarium graminearum (f.graminearum) and fusarium flavum (f.culmorum), is common in crops such as wheat, barley, oats, corn and the like, and is a trichothecene compound with wide distribution range and great influence. DON has strong toxicity to human bodies and animals, and is mainly reflected in influencing the normal growth and development of life bodies and causing potential harm to spleen, heart, liver and the like. DON is present in all countries around the world, is mainly distributed in the south of the Yangtze river in China, not only pollutes cereal crops and products thereof, but also can be transferred to animal-derived foods such as meat products, internal organs, milk, dairy products, eggs and the like. Therefore, it is necessary to develop a detection method having advantages such as high sensitivity, strong specificity, good stability, and low detection limit.
Electrochemiluminescence (ECL), also called Electrochemiluminescence, is a phenomenon of chemiluminescence directly or indirectly initiated by applying a certain voltage to an electrode to perform an electrochemical reaction; the electrochemiluminescence analysis is an analysis method for realizing quantitative determination of a substance to be detected according to a linear relation between the concentration of the substance to be detected and an ECL signal by collecting the ECL signal through an optical instrument such as a photomultiplier tube. Compared with other analysis technologies, the electrochemiluminescence sensor integrates the advantages of electrochemical detection and chemiluminescence, and has the excellent characteristics of high sensitivity, rapidness, low background and the like. The aptamer is an oligonucleotide chain selected from a DNA library by using a SELEX technology, has high specificity and affinity to a target substance, has a function similar to that of a traditional antibody, and is smaller in physical size, easier to modify and label and higher in stability. The aptamer is used as an identification element and is combined with the electrochemiluminescence technology, so that the detection performance of the electrochemiluminescence sensor can be improved. The invention prepares the electrochemiluminescence aptamer sensor for inhibiting the ruthenium-based metal organic framework based on the bismuth sulfide nanorod, and realizes high-sensitivity detection of vomitoxin.
Ruthenium-based metal organic framework (Ru-MOFs) compounds are ideal materials for electrochemiluminescence probes due to the unique sheet-shaped stacking structure, high luminous efficiency, good chemical and physical stability and biocompatibility. The Ru-MOFs nano material combines the advantages of bipyridyl ruthenium and a metal organic framework, so that the Ru-MOFs nano material has more excellent performance. In addition, Bi2S3As a direct band gap material having a band gap width of 1.3eV, it is widely used in the fields of photocatalysts, solar cells, hydrogen storage materials, thermoelectric materials, and the like.
Fang et al, TiO utilizing coumarin-containing silicon phthalocyanine and luminol2And (B) preparing an electrochemiluminescence immunosensor by using the integrated biological probe, and being used for quantitative analysis of vomitoxin. However, luminol as a luminescent reagent needs to be dissolved under a slightly alkaline condition so as to generate an effective electrochemiluminescence signal, and the condition limits the application of the luminol in an electrochemiluminescence immunosensor; lv and the like utilize gold nanoparticle functionalized nano porous cobalt to catalyze ruthenium-silicon spheres to prepare the electrochemiluminescence immunosensor for detecting vomitoxin. However, the synthesis method of the nano material used is complex and needs dangerous chemicals such as ammonia water. At present, no report is made on the detection of vomitoxin by an electrochemiluminescence sensor prepared by the two nano materials and an electrochemiluminescence aptamer sensor. Therefore, it is very important to prepare an electrochemiluminescence aptamer sensor which is simple in synthesis method, convenient to operate and free of various condition limitations for detecting vomitoxin.
Disclosure of Invention
The invention aims to prepare an electrochemiluminescence aptamer sensor based on bismuth sulfide nanorod inhibition ruthenium-based metal organic framework to realize high-sensitivity detection of vomitoxin.
The invention provides a method for detecting vomitoxin by an electrochemiluminescence aptamer sensor prepared by inhibiting a ruthenium-based metal organic framework based on a bismuth sulfide nanorod, which comprises the following steps of:
firstly, synthesizing a ruthenium-based metal organic framework (Ru-MOFs), namely mixing 9mg of tris (4, 4-dicarboxyl bipyridyl) ruthenium chloride, 45mg of zinc nitrate, 45mg of pyrazine, 90mg of hexadecyl trimethyl ammonium chloride and 45mL of pure water in a 100mL beaker, uniformly performing ultrasonic treatment, transferring the mixed solution into a reaction kettle with a polytetrafluoroethylene lining, reacting for 4 hours at 120 ℃ to obtain orange turbid liquid, centrifuging at 8500rpm for 10 minutes, and finally performing vacuum drying to obtain the ruthenium-based metal organic framework (Ru-MOFs);
second, bismuth sulfide (Bi)2S3) Weighing 1.82g of bismuth nitrate, adding 25mL of glycol, and deoxidizing for 15min in a nitrogen atmosphere to obtain a solution A; weighing 1.351g of sodium sulfide, adding 10mL of ethylene glycol and 20mL of pure water, and stirring until the sodium sulfide and the pure water are completely dissolved to obtain a solution B; dropwise adding the solution B into the solution A under vigorous stirring, then adding 1.922g of urea and 20ml of pure water, and continuing stirring for 30 min; transferring the mixed solution into a reaction kettle with a polytetrafluoroethylene lining, reacting for 12 hours at 180 ℃ to obtain black turbid liquid, washing for three times, and drying in vacuum to obtain bismuth sulfide (Bi)2S3)；
Mechanically polishing a Glassy Carbon Electrode (GCE) on a chamois paved with alumina powder, washing residual powder on the surface by using pure water, sequentially placing the polished glassy carbon electrode in ethanol and distilled water for ultrasonic treatment for 5-10min respectively, and drying at room temperature;
fourthly, transferring 6 mu L of gold nanoparticle (AuNPs) solution to be dripped on the surface of the glassy carbon electrode, and drying at room temperature to obtain AuNPs/GCE; continuously transferring 20 mu L of Ru-MOFs solution to be dropwise added on the surface of AuNPs/GCE, and drying at room temperature to obtain Ru-MOFs/AuNPs/GCE; then, soaking Ru-MOFs/AuNPs/GCE into 0.5-2.5 mu M/L sulfydryl functionalized vomitoxin aptamer solution (SH-Apt), incubating for 2h at 37 ℃, and washing the electrode by phosphate buffer solution with the pH of 7.4 to obtain an SH-Apt/Ru-MOFs/AuNPs modified glassy carbon electrode; finally, 5 microliter of MCH solution is transferred and dripped on the surface of SH-Apt/AuNPs/GCE to seal the nonspecific active sites on the surface of the electrode, and the electrode is stored in a refrigerator at 4 ℃ for later use;
fifthly, taking 0.5-2mg/mL bismuth sulfide solution10mL of HAuCl was added at 0.8mL and 0.1M/L4The solution and 2mL of 50mM/L sodium citrate solution are subjected to water bath at the temperature of 80 ℃ to synthesize Bi2S3@ Au complex, concussing it with aptamer complementary strand (cDNA) for 12h at room temperature to obtain Au @ Bi2S3-a cDNA tag;
and a sixth step: mixing Au @ Bi2S3Incubating the-cDNA tag and SH-Apt/AuNPs/GCE at 3-60 ℃ for 30-120min to obtain Au @ Bi2S3-a cDNA/SH-Apt/AuNPs modified electrode;
the seventh step: and immersing the modified electrode obtained in the sixth step into vomitoxin standard solutions with different concentrations, incubating for 40min in a refrigerator at 4 ℃, washing the surface of the electrode with pure water to obtain the electrogenerated chemiluminescence aptamer sensor for vomitoxin, and storing in the refrigerator at 4 ℃ for later use.
Preferably, the concentration of the thiol-functionalized vomitoxin aptamer solution in the fourth step is 1.0. mu.M/L.
Preferably, the concentration of the bismuth sulfide solution in the fifth step is 0.5 mg/mL.
Preferably, the incubation time in the sixth step is 60 minutes and the incubation temperature is 37 ℃.
The thiol-functionalized vomitoxin aptamer solution (SH-Apt) and the aptamer complementary strand (cDNA) in the present invention were purchased from Shanghai.
The invention also provides an application of the electrochemiluminescence aptamer sensor in detection of vomitoxin, which specifically comprises the following steps:
measuring by using a three-electrode system, taking the electrochemiluminescence aptamer sensor prepared by the method as a working electrode, taking Ag/AgCl as a reference electrode, taking a platinum wire electrode as a counter electrode, and testing in 0.1M/L phosphate buffer solution with pH of 7.4; the method is characterized in that the vomitoxin standard solutions with different concentrations are detected by adopting a potential range of-1.8V-1.5V, a scanning speed of 0.05V/s and an electrochemiluminescence device photomultiplier 800V, and the vomitoxin is detected by using the ECL signal intensity of 1.5V acquired by the electrochemiluminescence device and the relation between the ECL signal intensity and the vomitoxin standard solution concentration.
The invention providesProvides a preparation method of an electrochemiluminescence aptamer sensor prepared by inhibiting a ruthenium-based metal organic framework based on a bismuth sulfide nanorod and high-sensitivity detection of vomitoxin, and is characterized in that the electrochemiluminescence aptamer sensor is used as an electrochemiluminescence probe and further functionalizes the vomitoxin aptamer based on the large specific surface area and high luminous efficiency of Ru-MOFs nano materials, and meanwhile, Bi is used2S3Labeling the aptamer complementary strand, the aptamer binding to the complementary strand such that Bi2S3Acts on the Ru-MOFs nano material to inhibit the ECL signal thereof, and realizes the high-sensitivity and high-stability detection of the vomitoxin by utilizing the specific recognition of the aptamer to the vomitoxin. The prepared vomitoxin electrochemiluminescence aptamer sensor has the advantages of strong specificity, high sensitivity, good stability, low detection limit and the like, can be used for detecting vomitoxin in food, and has very important application value in the aspect of food safety monitoring.
Drawings
FIG. 1 is an SEM image of Ru-MOFs in an example of the present invention;
FIG. 2 shows Bi in example of the present invention2S3SEM picture of (1);
FIG. 3 shows Bi of different concentrations in the preparation of an electrochemiluminescence aptamer sensor according to an embodiment of the invention2S3Influence on the intensity of Ru-MOFs electrochemiluminescence signals;
FIG. 4 is a graph of the effect of aptamer concentration on ECL signal response during the preparation of an electrochemiluminescence aptamer sensor in an embodiment of the invention;
FIG. 5 is a graph showing the effect of incubation time on ECL signal response during the preparation of an electrochemiluminescence aptamer sensor according to an embodiment of the invention;
FIG. 6 is a graph showing the effect of incubation temperature on ECL signal response during the preparation of an electrochemiluminescence aptamer sensor according to an embodiment of the invention.
Detailed Description
The following is a description of specific embodiments of the present invention: the present embodiment is implemented on the premise of the technical solution of the present invention, and a detailed implementation manner and a specific operation flow are given, but the application scope of the present invention is not limited to the following embodiments.
Example 1:
firstly, synthesizing Ru-MOFs, namely mixing 9mg of tris (4, 4-dicarboxyl bipyridyl) ruthenium chloride, 45mg of zinc nitrate, 45mg of pyrazine, 90mg of hexadecyl trimethyl ammonium chloride and 45mL of pure water in a 100mL beaker, uniformly performing ultrasonic treatment, transferring the mixed solution into a reaction kettle with a polytetrafluoroethylene lining, reacting for 4 hours at 120 ℃ to obtain orange turbid liquid, centrifuging at 8500rpm for 10 minutes, and finally performing vacuum drying to obtain the Ru-MOFs.
And secondly, synthesizing bismuth sulfide, namely weighing 1.82g of bismuth nitrate, adding 25mL of ethylene glycol, and deoxidizing for 15min in a nitrogen atmosphere to obtain a solution A. 1.351g of sodium sulfide was weighed, 10mL of ethylene glycol and 20mL of pure water were added, and the mixture was stirred until completely dissolved to obtain a solution B. Solution B was added dropwise to solution A with vigorous stirring, followed by addition of 1.922g of urea and 20ml of pure water, and stirring was continued for 30 min. And transferring the mixed solution into a reaction kettle with a polytetrafluoroethylene lining, reacting for 12 hours at 180 ℃ to obtain black turbid liquid, washing for three times, and then carrying out vacuum drying to obtain bismuth sulfide.
And thirdly, mechanically polishing and polishing the Glassy Carbon Electrode (GCE) on a chamois paved with alumina powder, washing residual powder on the surface by using pure water, sequentially placing the polished glassy carbon electrode in ethanol and distilled water for ultrasonic treatment for 5-10min respectively, and drying at room temperature.
And fourthly, transferring 6 mu L of prepared gold nanoparticle (AuNPs) solution to be dripped on the surface of the glassy carbon electrode, and drying at room temperature to obtain AuNPs/GCE. And continuously transferring 20 mu L of Ru-MOFs solution to be dropwise added on the surface of AuNPs/GCE, and drying at room temperature to obtain the Ru-MOFs/AuNPs/GCE. In order to determine the optimal use concentration of the aptamer, the Ru-MOFs/AuNPs/GCE is immersed in thiol-functionalized vomitoxin aptamer solutions with the concentrations of 0.5, 1.0, 1.5, 2.0 and 2.5 mu M/L respectively, the solutions are incubated for 2 hours at 37 ℃, and the electrode is washed by phosphate buffer solution with the pH of 7.4 to obtain the SH-Apt/Ru-MOFs/AuNPs modified glassy carbon electrode. Finally, 5 microliter of MCH solution is transferred and dripped on the surface of SH-Apt/AuNPs/GCE to seal the nonspecific active sites on the surface of the electrode, and the electrode is stored in a refrigerator at 4 ℃ for later use.
The fifth step is to get0.5mg/mL bismuth sulfide solution 10mL, 0.8mL0.1M/L HAuCl was added4The solution and 2mL of 50mM/L sodium citrate solution are used for synthesizing Bi under the condition of 80 ℃ water bath2S3@ Au complex, concussing it with aptamer complementary strand (cDNA) for 12h at room temperature to obtain Au @ Bi2S3-a cDNA tag.
And a sixth step: mixing Au @ Bi2S3Incubating the-cDNA marker with SH-Apt/AuNPs/GCE at 37 ℃ for 1h to obtain Au @ Bi2S3-cDNA/SH-Apt/AuNPs modified electrode.
The seventh step: and immersing the modified electrode obtained in the sixth step into vomitoxin standard solutions with different concentrations, incubating for 40min in a refrigerator at 4 ℃, washing the surface of the electrode with pure water to obtain the electrogenerated chemiluminescence aptamer sensor for vomitoxin, and storing in the refrigerator at 4 ℃ for later use.
And eighthly, measuring by using a three-electrode system, taking the prepared electrochemiluminescence aptamer sensor as a working electrode, taking Ag/AgCl as a reference electrode and taking a platinum wire electrode as a counter electrode, and testing in 0.1M/L phosphate buffer solution with pH of 7.4. The potential range is-1.8V-1.5V, the scanning speed is 0.05V/s, the electrochemiluminescence device photomultiplier 800V is used for detecting vomitoxin standard solutions with different concentrations, and a working curve is drawn according to the ECL signal intensity of 1.5V acquired by the electrochemiluminescence device and the relation between the ECL signal intensity and the vomitoxin standard solution concentration.
Example 2:
first step, as in example 1
Second step, as in example 1
Third step, as in example 1
The fourth step, as in example 1, where the concentration of the thiol-functionalized vomitoxin aptamer solution was chosen to be 1.0. mu.M/L.
Fifthly, in order to verify the feasibility of the experiment and show that different concentrations of bismuth sulfide have different influences on the ECL signals of the Ru-MOFs, 10mL of 0, 0.5, 1.0, 1.5 and 2mg/mL bismuth sulfide solution is respectively taken, and 0.8mL of 0.1M/L HAuCl is added4The solution and 2mL of 50mM/L sodium citrate solution are used for synthesizing Bi under the condition of 80 ℃ water bath2S3@ Au complex, shaking the complex with aptamer complementary strand (cDNA) at a concentration of 100. mu.M/L for 12h at room temperature to obtain Au @ Bi2S3-a cDNA tag.
Sixth step, as in example 1
Seventh step, as in example 1
Eighth step, as in example 1
Example 3:
first step, as in example 1
Second step, as in example 1
Third step, as in example 1
The fourth step, as in example 1, where the concentration of the thiol-functionalized vomitoxin aptamer solution was chosen to be 1.0. mu.M/L.
Fifth step, as in example 1
Sixthly, in order to obtain the optimal incubation time, Au @ Bi2S3Incubating the-cDNA marker and SH-Apt/AuNPs/GCE at 37 ℃ for 30, 60, 90 and 120 minutes to obtain Au @ Bi2S3-cDNA/SH-Apt/AuNPs modified electrode.
Seventh step, as in example 1
Eighth step, as in example 1
Example 4:
first step, as in example 1
Second step, as in example 1
Third step, as in example 1
The fourth step, as in example 1, where the concentration of the thiol-functionalized vomitoxin aptamer solution was chosen to be 1.0. mu.M/L.
Fifth step, as in example 1
Sixthly, in order to determine the optimal incubation temperature, Au @ Bi2S3Incubating the-cDNA marker and SH-Apt/AuNPs/GCE for 1h at the temperature of 4, 25, 37 and 60 ℃ respectively to obtain Au @ Bi2S3-cDNA/SH-Apt/AuNPs modified electrode.
Seventh step, as in example 1
Eighth step, as in example 1
As shown in the attached figure 1, the SEM image of Ru-MOFs has a unique sheet-shaped stacking structure and has a large specific surface area, so that the Ru-MOFs can be used as an effective electrochemiluminescence probe.
As shown in FIG. 2, Bi2S3The SEM picture of (A) is of a rod-shaped structure, has uniform size and regular distribution, and provides more binding sites for binding complementary DNA strands.
As shown in FIG. 3, it can be seen that Bi is associated with2S3The concentration is increased, the electrochemiluminescence signal intensity of Ru-MOFs at the anode is gradually weakened, the electrochemiluminescence signal intensity at the cathode is increased, and the feasibility of experimental design is proved.
As shown in FIG. 4, it can be seen that when the aptamer concentration is increased to 1.0 μ M/L, the electrochemiluminescence signal is the highest, and the subsequent binding of the target substance to be detected is affected by the too high aptamer concentration, so that 1.0 μ M/L is selected as the optimal aptamer concentration.
As shown in FIG. 5, when the time reaches 60 minutes, the labeled complementary DNA strand is completely bound to the aptamer to a saturated state, the electrochemiluminescence signal intensity is the highest, so 60 minutes is selected as the optimal incubation time.
As shown in FIG. 6, when the temperature is 37 ℃, the electrochemiluminescence signal intensity is the highest, and the structural stability of the aptamer and the complementary DNA strand is affected by overhigh temperature, so that 37 ℃ is selected as the optimal incubation temperature.
In conclusion, in the preparation process of the electrochemiluminescence aptamer sensor, when the concentration of the vomitoxin as the detection target substance is 0.5mg/mL, the preparation method has the following key factors: the aptamer concentration, incubation time and incubation temperature are optimized, and the performance detection result of the sensor under the optimal condition is as follows:
the result of condition optimization shows that when the aptamer concentration in the system is 1.0 mu M/L, the incubation time is 60 minutes, and the incubation temperature is 37 ℃, the obtained vomitoxin electrochemiluminescence aptamer sensor has the best performance.
The electrochemiluminescence aptamer sensor is prepared based on the intensity of electrochemiluminescence signals of bismuth sulfide nanorods for inhibiting Ru-MOFs, high-sensitivity detection of vomitoxin is achieved, and compared with other analysis technologies, the electrochemiluminescence aptamer sensor prepared by the method is high in sensitivity and good in specificity. The scheme expands the application of the electrochemiluminescence biosensor in the aspect of food safety and provides a new idea for detecting pollutants in food.
Claims (6)
1. A preparation method of an electrochemiluminescence aptamer sensor based on a bismuth sulfide nanorod for inhibiting a ruthenium-based metal organic framework comprises the following steps:
firstly, synthesizing a ruthenium-based metal organic framework (Ru-MOFs), namely mixing 9mg of tris (4, 4-dicarboxyl bipyridyl) ruthenium chloride, 45mg of zinc nitrate, 45mg of pyrazine, 90mg of hexadecyl trimethyl ammonium chloride and 45mL of pure water in a 100mL beaker, uniformly performing ultrasonic treatment, transferring the mixed solution into a reaction kettle with a polytetrafluoroethylene lining, reacting for 4 hours at 120 ℃ to obtain orange turbid liquid, centrifuging at 8500rpm for 10 minutes, and finally performing vacuum drying to obtain the ruthenium-based metal organic framework (Ru-MOFs);
second, bismuth sulfide (Bi)2S3) Weighing 1.82g of bismuth nitrate, adding 25mL of glycol, and deoxidizing for 15min in a nitrogen atmosphere to obtain a solution A; weighing 1.351g of sodium sulfide, adding 10mL of ethylene glycol and 20mL of pure water, and stirring until the sodium sulfide and the pure water are completely dissolved to obtain a solution B; dropwise adding the solution B into the solution A under vigorous stirring, then adding 1.922g of urea and 20ml of pure water, and continuing stirring for 30 min; transferring the mixed solution into a reaction kettle with a polytetrafluoroethylene lining, reacting for 12 hours at 180 ℃ to obtain black turbid liquid, washing for three times, and drying in vacuum to obtain bismuth sulfide (Bi)2S3)；
Mechanically polishing a Glassy Carbon Electrode (GCE) on a chamois paved with alumina powder, washing residual powder on the surface by using pure water, sequentially placing the polished glassy carbon electrode in ethanol and distilled water for ultrasonic treatment for 5-10min respectively, and drying at room temperature;
fourthly, transferring 6 mu L of gold nanoparticle (AuNPs) solution to be dripped on the surface of the glassy carbon electrode, and drying at room temperature to obtain AuNPs/GCE; continuously transferring 20 mu L of Ru-MOFs solution to be dropwise added on the surface of AuNPs/GCE, and drying at room temperature to obtain Ru-MOFs/AuNPs/GCE; then, soaking Ru-MOFs/AuNPs/GCE into 0.5-2.5 mu M/L sulfydryl functionalized vomitoxin aptamer (SH-Apt) solution, incubating for 2h at 37 ℃, and washing the electrode by using phosphate buffer solution with the pH of 7.4 to obtain an SH-Apt/Ru-MOFs/AuNPs modified glassy carbon electrode; finally, 5 mu L of 6-sulfydryl-1-hexanol (MCH) solution is transferred and dripped on the surface of SH-Apt/AuNPs/GCE to seal the nonspecific active sites on the surface of the electrode, and the electrode is stored in a refrigerator at 4 ℃ for later use;
fifthly, taking 10mL of 0.5-2mg/mL bismuth sulfide solution, adding 0.8mL of 0.1M/L HAuCl4The solution and 2mL of 50mM/L sodium citrate solution are subjected to water bath at the temperature of 80 ℃ to synthesize Bi2S3@ Au complex, concussing it with aptamer complementary strand (cDNA) for 12h at room temperature to obtain Au @ Bi2S3-a cDNA tag;
and a sixth step: mixing Au @ Bi2S3Incubating the-cDNA tag and SH-Apt/AuNPs/GCE at 3-60 ℃ for 30-120min to obtain Au @ Bi2S3-a cDNA/SH-Apt/AuNPs modified electrode;
the seventh step: and immersing the modified electrode obtained in the sixth step into vomitoxin standard solutions with different concentrations, incubating for 40min in a refrigerator at 4 ℃, washing the surface of the electrode with pure water to obtain the electrogenerated chemiluminescence aptamer sensor for vomitoxin, and storing in the refrigerator at 4 ℃ for later use.
2. The method of claim 1, wherein: the concentration of the thiol-functionalized emetic toxin aptamer solution in the fourth step was 1.0. mu.M/L.
3. The method of claim 1, wherein: the concentration of the bismuth sulfide solution in the fifth step was 0.5 mg/mL.
4. The method of claim 1, wherein: in the sixth step, the incubation time is 60min and the incubation temperature is 37 ℃.
5. An electrochemiluminescence aptamer sensor for inhibiting a ruthenium-based metal organic framework based on bismuth sulfide nanorods, which is characterized by being prepared by the method of claim 1.
6. Use of an electrochemiluminescent aptamer sensor according to claim 5 for detecting vomitoxin, wherein:
measuring by using a three-electrode system, taking an electrochemiluminescence aptamer sensor as a working electrode, Ag/AgCl as a reference electrode and a platinum wire electrode as a counter electrode, and testing in 0.1M/L, pH 7.4.4 phosphate buffer solution; the method is characterized in that the vomitoxin standard solutions with different concentrations are detected by adopting a potential range of-1.8V-1.5V, a scanning speed of 0.05V/s and an electrochemiluminescence device photomultiplier 800V, and the vomitoxin is detected by using the ECL signal intensity of 1.5V acquired by the electrochemiluminescence device and the relation between the ECL signal intensity and the vomitoxin standard solution concentration.
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| CN114878652A (en) * | 2022-05-23 | 2022-08-09 | 集美大学 | Preparation method and application of cathode electrochemiluminescence aptamer sensor |
| CN114878652B (en) * | 2022-05-23 | 2023-10-20 | 集美大学 | Preparation method and application of cathode electrochemiluminescence aptamer sensor |
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