Detection electrode assembly and electrochemical biosensor

Abstract

The utility model relates to the technical field of electronic component carriers, in particular to a detection electrode assembly and an electrochemical biosensor, the detection electrode assembly is used for the electrochemical biosensor, the detection electrode assembly comprises a substrate layer and an electrode layer, the electrode layer is arranged on the substrate layer, and the electrode layer is arranged on the substrate layer. The electrode layer comprises a detection electrode and a calibration electrode; wherein during electrochemical detection, the detection electrode responds to a biological material to be detected to generate a detection identification signal, and the calibration electrode responds to a reference biological material to generate a reference identification signal.

Description

Technical Field

[0001] The present disclosure relates to the field of biosensor technology, and in particular to a detection electrode assembly, a detection method, and an electrochemical biosensor. Background Art

[0002] With the development and advancement of technology, electrochemical biosensors are increasingly being used in medical diagnosis, environmental monitoring, food safety and other fields. During use, the performance of electrochemical biosensors can degrade due to environmental interference and background noise, and their performance can also change with aging, resulting in a decrease in the accuracy of electrochemical biosensor detection results.

[0003] It should be noted that the information disclosed in the above background technology section is only used to enhance the understanding of the background of the present disclosure, and therefore may include information that does not constitute prior art known to ordinary technicians in the field. Summary of the Invention

[0004] The present disclosure aims to provide a detection electrode assembly and an electrochemical biosensor, thereby improving the accuracy of the detection results of the electrochemical biosensor to at least a certain extent.

[0005] According to a first aspect of the present disclosure, there is provided a detection electrode assembly, wherein the detection electrode assembly is used for an electrochemical biosensor, and the detection electrode assembly comprises:

[0006] basal layer;

[0007] An electrode layer is provided on the base layer, and the electrode layer includes a detection electrode and a calibration electrode; wherein, during electrochemical detection, the detection electrode is used to generate a detection identification signal in response to the biomaterial to be detected, and the calibration electrode is used to generate a reference identification signal in response to the reference biomaterial.

[0008] According to one embodiment of the present disclosure, the electrode layer further includes a counter electrode and a reference electrode, the detection electrode and the counter electrode form a first polarization circuit, the calibration electrode and the counter electrode form a second polarization circuit, and the reference electrode is used to provide a reference potential.

[0009] According to one embodiment of the present disclosure, a preset electrode area is provided on the base layer, the detection electrode and the calibration electrode are provided in the preset electrode area, and the counter electrode at least partially surrounds the preset electrode area.

[0010] According to one embodiment of the present disclosure, the detection electrode assembly further includes:

[0011] A conductive layer is provided with a first wire, a second wire, a third wire and a fourth wire, the first wire is connected to the detection electrode, the second wire is connected to the calibration electrode, the third wire is connected to the reference electrode, and the fourth wire is connected to the counter electrode.

[0012] According to one embodiment of the present disclosure, the detection electrode assembly further includes:

[0013] An insulating layer is provided on the base layer, the insulating layer covers the conductive layer, and a detection opening is provided on the insulating layer, and the orthographic projections of the detection electrode, the calibration electrode, the reference electrode and the counter electrode on the insulating layer are located in the detection opening.

[0014] According to one embodiment of the present disclosure, a biometric recognition element is provided on each of the detection electrode and the calibration electrode. The biometric recognition element is used to recognize a preset biomaterial and generate a recognition signal.

[0015] According to one embodiment of the present disclosure, the detection electrode is used to detect kanamycin, and the biorecognition element is a DNA aptamer of kanamycin.

[0016] According to a second aspect of the present disclosure, a detection method is provided for the above-mentioned detection electrode assembly, the detection method comprising:

[0017] Acquiring a detection identification signal generated by the detection electrode in response to a biomaterial to be detected, wherein the biomaterial to be detected contains a detection target;

[0018] Acquiring a reference identification signal generated by the calibration electrode in response to a reference biological material, wherein the reference biological material contains a detection target, and the detection target content in the reference biological material is a critical value;

[0019] The detection identification signal is compared with the reference identification signal to determine whether the content of the detection target in the detection biological material is qualified.

[0020] According to a third aspect of the present disclosure, an electrochemical biosensor is provided, comprising the above-mentioned detection electrode assembly.

[0021] According to one embodiment of the present disclosure, the electrochemical biosensor further includes:

[0022] A detection circuit is connected to the detection electrode and the calibration electrode respectively, and is used to form a reference curve according to the reference identification signal and to form a detection curve according to the detection identification signal.

[0023] The detection electrode provided by the embodiment of the present disclosure includes a base layer and an electrode layer, and the electrode layer is arranged on the base layer. The electrode layer includes a detection electrode and a calibration electrode. During electrochemical detection, the detection electrode generates a detection identification signal in response to the biological material to be detected, and the calibration electrode generates a reference identification signal in response to the reference biological material. Since the detection electrode and the calibration electrode are located in the same environment, and their aging degrees are similar as the electrochemical biosensor is used, the detection result is determined by comparing the reference identification signal and the detection identification signal, which is not affected by the environment and the aging of the detection electrode, thereby improving the detection accuracy of the electrochemical biosensor.

[0024] It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The accompanying drawings are incorporated into and constitute a part of the specification, illustrate embodiments consistent with the present disclosure, and together with the specification, are used to explain the principles of the present disclosure. Obviously, the drawings described below are only some embodiments of the present disclosure, and those skilled in the art can derive other drawings based on these drawings without inventive effort.

[0026] Figure 1 A schematic diagram of a detection electrode assembly provided by an exemplary embodiment of the present disclosure;

[0027] Figure 2 A schematic diagram of another detection electrode assembly provided by an exemplary embodiment of the present disclosure;

[0028] Figure 3 A schematic diagram of an insulating layer provided by an exemplary embodiment of the present disclosure;

[0029] Figure 4 A flow chart of a detection method provided for an exemplary embodiment of the present disclosure;

[0030] Figure 5 A curve chart of a detection result provided by an exemplary embodiment of the present disclosure;

[0031] Figure 6 A curve chart of another detection result provided by an exemplary embodiment of the present disclosure.

[0032] Description of reference numerals:

[0033] 10. Base layer; 21. Detection electrode; 22. Calibration electrode; 23. Reference electrode; 24. Counter electrode; 31. First wire; 32. Second wire; 33. Third wire; 34. Fourth wire; 40. Insulation layer; 41. Detection opening. DETAILED DESCRIPTION

[0034] Example embodiments will now be described more fully with reference to the accompanying drawings. However, example embodiments can be implemented in many forms and should not be construed as limited to the examples set forth herein; rather, these embodiments are provided so that this disclosure will be comprehensive and complete and will fully convey the concepts of the example embodiments to those skilled in the art. The described features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. In the following description, many specific details are provided to provide a full understanding of the embodiments of the present disclosure. However, those skilled in the art will appreciate that the technical solutions of the present disclosure may be practiced without one or more of the specific details, or that other methods, materials, devices, etc. may be employed. In other cases, well-known technical solutions are not shown or described in detail to avoid obscuring various aspects of the present disclosure.

[0035] In addition, the drawings are merely schematic illustrations of the present disclosure and are not necessarily drawn to scale. Like reference numerals in the drawings represent identical or similar parts, and thus their repeated descriptions will be omitted. The terms "a," "an," and "the" are used to indicate the presence of one or more elements; the terms "including" and "having" are used to indicate an open-ended inclusive meaning and indicate that additional elements may be present in addition to the listed elements.

[0036] The exemplary embodiment of the present disclosure first provides a detection electrode assembly, which is used for an electrochemical biosensor, such as Figure 1 As shown, the detection electrode assembly includes: a base layer 10 and an electrode layer. The electrode layer is provided on the base layer 10, and the electrode layer includes a detection electrode 21 and a calibration electrode 22. During electrochemical detection, the detection electrode 21 is used to generate a detection identification signal in response to the biomaterial to be detected, and the calibration electrode 22 is used to generate a reference identification signal in response to the reference biomaterial.

[0037] The detection electrode assembly provided by the embodiment of the present disclosure includes a base layer 10 and an electrode layer. The electrode layer is arranged on the base layer 10. The electrode layer includes a detection electrode 21 and a calibration electrode 22. During electrochemical detection, the detection electrode 21 generates a detection identification signal in response to the biological material to be detected, and the calibration electrode 22 generates a reference identification signal in response to the reference biological material. Since the detection electrode 21 and the calibration electrode 22 are located in the same environment, and their aging degrees are similar as the electrochemical biosensor is used, the detection result is determined by comparing the reference identification signal and the detection identification signal, which is not affected by the environment and the aging of the detection electrode 21, thereby improving the detection accuracy of the electrochemical biosensor.

[0038] The following is a detailed description of the detection electrode assembly provided in the embodiment of the present disclosure:

[0039] The base layer 10 supports the structure of the detection electrode 21 and is made of an insulating material. The base layer 10 can be made of a rigid or flexible material. For example, the base layer 10 can be made of a flexible material such as polyethylene terephthalate (PET) or polyimide (PI). The flexible base layer 10 improves the adaptability of the detection electrode 21, making it easier to use in different scenarios.

[0040] The electrode layer is arranged on the base layer 10, and the electrode layer includes a detection electrode 21 and a calibration electrode 22. During electrochemical detection, the detection electrode 21 generates a detection identification signal in response to the biological material to be detected, and the calibration electrode 22 generates a reference identification signal in response to the reference biological material. The biological material to be detected and the reference biological material are the same biological materials. The biological material to be detected contains a target detection object, for example, the biological material to be detected is milk, and the target detection object is kanamycin. The reference biological material contains a target detection object, and the content of the target detection object in the reference biological material is a critical value. The critical value can be understood as when the content of the target detection object in the biological material to be detected and the critical value meet a preset relationship (for example, greater than, less than or equal to, etc.), it is qualified. For example, the reference biological material is milk, and the content of kanamycin in the reference biological material is the critical value specified by the national standard.

[0041] It should be noted that the biological material to be detected in the embodiment of the present disclosure is not limited to milk, that is, the detection electrode 21 can not only detect milk and other dairy products, but can also be used to detect blood, urine, saliva, etc.

[0042] Furthermore, the electrode layer also includes a counter electrode 24 and a reference electrode 23. The detection electrode 21 and the counter electrode 24 form a first polarization circuit, the calibration electrode 22 and the counter electrode 24 form a second polarization circuit, and the reference electrode 23 is used to provide a reference potential.

[0043] During the detection process, the detection electrode 21 and the reference electrode 23 form a first measurement circuit, and the detection identification signal can be an electrical signal in the first measurement circuit, or a processed electrical signal in the first measurement circuit. The calibration electrode 22 and the reference electrode 23 form a second measurement circuit, and the reference identification signal can be an electrical signal in the second measurement circuit, or a processed electrical signal in the second measurement circuit.

[0044] The detection electrode 21, calibration electrode 22, and counter electrode 24 can be graphite electrodes, and the reference electrode 23 can be a silver electrode or a silver chloride electrode. The wires in the conductive layer can be silver wires. Of course, in practical applications, the electrodes and wires can also be made of other conductive materials, and the embodiments of the present disclosure are not limited thereto.

[0045] The base layer 10 has a predetermined electrode region, with the detection electrode 21 and the calibration electrode 22 disposed in the predetermined electrode region, and the counter electrode 24 at least partially surrounding the predetermined electrode region. For example, the detection electrode 21 and the calibration electrode 22 may be elliptical electrodes, with a gap between them. For example, the counter electrode 24 may have a ring-shaped or nearly ring-shaped structure, surrounding the detection electrode 21 and the calibration electrode 22. The reference electrode 23 may also be disposed within the ring-shaped structure of the counter electrode 24, with the ring-shaped counter electrode 24 having an opening, and the reference electrode 23 disposed near the opening.

[0046] Further, such as Figure 2 As shown, the detection electrode assembly may further include a conductive layer, in which a first wire 31, a second wire 32, a third wire 33, and a fourth wire 34 are provided. The first wire 31 is connected to the detection electrode 21, the second wire 32 is connected to the calibration electrode 22, the third wire 33 is connected to the reference electrode 23, and the fourth wire 34 is connected to the counter electrode 24. The electrodes are led out through the wires in the conductive layer, and the ends of the wires facing away from the electrodes are used to connect to an external circuit.

[0047] Further, such as Figure 3 As shown, the detection electrode assembly further includes an insulating layer 40, which is disposed on the base layer 10 and covers the conductive layer. A detection opening 41 is provided on the insulating layer 40, and the orthographic projections of the detection electrode 21, calibration electrode 22, reference electrode 23, and counter electrode 24 on the insulating layer 40 are located in the detection opening 41. That is, the detection electrode 21, calibration electrode 22, reference electrode 23, and counter electrode 24 are exposed to the detection opening 41, and the biomaterial and electrolyte are injected through the detection opening 41.

[0048] The detection electrode 21 and the calibration electrode 22 are respectively provided with a biorecognition element (bioprobe), which is used to identify a predetermined biomaterial and generate an identification signal. The bioprobe on the detection electrode 21 is the same as the bioprobe on the calibration electrode 22.

[0049] For example, when the detection electrode 21 is used to detect kanamycin, the biorecognition element is a kanamycin DNA aptamer, and the modified base sequence of the DNA aptamer is: 5′-HS-(CH 2 ) 6 -TGGGGGTTGAGGCTAAGCCGA-3′.

[0050] Gold particles of about 10 nm are sputtered on the detection electrode 21 and the calibration electrode 22, and the aptamer is fixed on the surface of the detection electrode 21 and the calibration electrode 22 by using gold-sulfur (Au-S) bonds, which effectively improves the stability of the sensor.

[0051] Optionally, the detection electrode 21 and the calibration electrode 22 are elliptical, with a major semi-axis of 2.1 mm and a minor semi-axis of 1.3 mm. The spacing between the detection electrode 21 and the calibration electrode 22 is 6.6 mm. The spacing between the wires in the conductive layer is 3.2 mm.

[0052] The preparation method of the detection electrode assembly provided in the embodiment of the present disclosure is as follows:

[0053] The base layer 10 (PET film or PI film) is ultrasonically cleaned in ultrapure water and ethanol in sequence, each ultrasonic cleaning is performed for a first preset time t1 (for example, 15 minutes), for a total of 2t1 (30 minutes), and then dried for later use;

[0054] Clean the customized screen with screen washing water to ensure that the screen is clean and not blocked, and dry it for later use; stir the conductive carbon paste, silver chloride paste and conductive silver paste for a second preset time (for example, 2 hours), place the cleaned screen on the base layer 10 and align them, then mix the conductive carbon paste evenly and pour it on the screen, print it with an automatic screen printer and dry it in a blast drying oven at a first preset temperature (for example, 120°C) for a third preset time (for example, 30 minutes) to obtain a detection electrode 21, a calibration electrode 22 and a counter electrode 24 for later use; use the same method as above to print a reference electrode 23 (Ag / AgCl) and a contact wire (Ag); clean the obtained detection electrode 21 with clean water or buffer solution (0.1M PBS buffer solution), and then dry it at a second preset temperature (for example, 60°C) for later use;

[0055] Adjust the magnetron sputtering process parameters to sputter gold particles of a preset thickness (about 10 nm) onto the detection electrode 21 and the calibration electrode 22; wash the magnetron sputtered electrode sheets with ultrapure water, dry them at room temperature and set aside.

[0056] When the detection electrode 21 is used to detect kanamycin, a kanamycin aptamer may be formed on the detection electrode 21 and the calibration electrode 22 as follows:

[0057] The test tube containing the kanamycin aptamer is centrifuged at a first preset speed (e.g., 5000 rpm) for a fourth preset time (e.g., 10 minutes). The test tube is opened and a first preset amount (e.g., 100 mL) of DEPC solution is added to prepare the aptamer solution at a preset concentration (e.g., 100 μmol / L) for later use. A second preset amount (e.g., 30 μL) of 1 μmol / L aptamer solution is dripped onto the cleaned detection electrode 21 and calibration electrode 22. The solution is incubated at a third preset temperature (e.g., 40°C) for a fifth preset time (e.g., 2 hours). The kanamycin aptamer that is not firmly adsorbed on the surface is then washed with water and a buffer solution. The base sequence of the modified DNA aptamer is: 5′-HS-(CH2)6-TGGGGGTTGAGGCTAAGCCGA-3′.

[0058] Optionally, the thickness of the magnetron sputtered gold particles is 10 nm to 20 nm. The buffer solution is PBS buffer (phosphate buffer) with a concentration of 0.1 M and a pH of approximately 7.4. The electrochemical performance test is a differential pulse voltammetry (DPV) test with a voltage range of -0.2 V to +0.6 V, a potential increment of 0.01 V, and an amplitude of 50 mV. The electrolyte solution for the electrochemical test is 0.1 M KCl in 0.1 M PBS, 5 mM [Fe(CN)6] 4- / 3- .

[0059] The detection electrode assembly provided by the embodiment of the present disclosure includes a base layer 10 and an electrode layer. The electrode layer is arranged on the base layer 10. The electrode layer includes a detection electrode 21 and a calibration electrode 22. During electrochemical detection, the detection electrode 21 generates a detection identification signal in response to the biological material to be detected, and the calibration electrode 22 generates a reference identification signal in response to the reference biological material. Since the detection electrode 21 and the calibration electrode 22 are located in the same environment, and their aging degrees are similar as the electrochemical biosensor is used, the detection result is determined by comparing the reference identification signal and the detection identification signal, which is not affected by the environment and the aging of the detection electrode 21, thereby improving the detection accuracy of the electrochemical biosensor.

[0060] The exemplary embodiments of the present disclosure also provide a detection method for the above-mentioned detection electrode assembly, such as Figure 4 As shown, the detection method includes the following steps:

[0061] Step S410, obtaining a detection identification signal generated by the detection electrode 21 in response to the biomaterial to be detected, where the biomaterial to be detected contains a detection target;

[0062] Step S420, obtaining a reference identification signal generated by the calibration electrode 22 in response to a reference biological material, wherein the reference biological material contains a detection target and the content of the detection target in the reference biological material is a critical value;

[0063] Step S430 : comparing the detection identification signal with the reference identification signal to determine whether the content of the detection target in the detection biological material is qualified.

[0064] The detection method provided by the embodiment of the present disclosure obtains a detection identification signal generated by the detection electrode 21 in response to the biological material to be detected and a reference identification signal generated by the calibration electrode 22 in response to the reference biological material, and determines whether the content of the detection target in the detection biological material is qualified based on the detection identification signal and the reference identification signal. Since the detection electrode 21 and the calibration electrode 22 are in the same environment and their aging degrees are similar as the electrochemical biosensor is used, the detection result is determined by comparing the reference identification signal and the detection identification signal, which is not affected by the environment and the aging of the detection electrode 21, thereby improving the detection accuracy of the electrochemical biosensor.

[0065] In the embodiment of the present disclosure, before step S410, the following steps may be included:

[0066] The base layer 10 (PET film or PI film) is ultrasonically cleaned in ultrapure water and ethanol in sequence, each ultrasonic cleaning is performed for a first preset time t1 (for example, 15 minutes), for a total of 2t1 (30 minutes), and then dried for later use;

[0067] Clean the customized screen with screen washing water to ensure that the screen is clean and not blocked, and dry it for later use; stir the conductive carbon paste, silver chloride paste and conductive silver paste for a second preset time (for example, 2 hours), place the cleaned screen on the base layer 10 and align them, then mix the conductive carbon paste evenly and pour it on the screen, print it with an automatic screen printer and dry it in a blast drying oven at a first preset temperature (for example, 120°C) for a third preset time (for example, 30 minutes) to obtain a detection electrode 21, a calibration electrode 22 and a counter electrode 24 for later use; use the same method as above to print a reference electrode 23 (Ag / AgCl) and a contact wire (Ag); clean the obtained detection electrode 21 with clean water or buffer solution (0.1M PBS buffer solution), and then dry it at a second preset temperature (for example, 60°C) for later use;

[0068] Adjust the magnetron sputtering process parameters to sputter gold particles of a preset thickness (about 10 nm) onto the detection electrode 21 and the calibration electrode 22; wash the magnetron sputtered electrode sheets with ultrapure water, dry them at room temperature and set aside.

[0069] When the detection electrode 21 is used to detect kanamycin, a kanamycin aptamer may be formed on the detection electrode 21 and the calibration electrode 22 as follows:

[0070] The test tube containing the kanamycin aptamer is centrifuged at a first preset speed (e.g., 5000 rpm) for a fourth preset time (e.g., 10 minutes). The test tube is opened and a first preset amount (e.g., 100 mL) of DEPC solution is added to prepare the aptamer solution at a preset concentration (e.g., 100 μmol / L) for later use. A second preset amount (e.g., 30 μL) of 1 μmol / L aptamer solution is dripped onto the cleaned detection electrode 21 and calibration electrode 22. The solution is incubated at a third preset temperature (e.g., 40°C) for a fifth preset time (e.g., 2 hours). The kanamycin aptamer that is not firmly adsorbed on the surface is then washed with water and a buffer solution. The base sequence of the modified DNA aptamer is: 5′-HS-(CH2)6-TGGGGGTTGAGGCTAAGCCGA-3′.

[0071] It should be noted that when the detection electrode 21 is used to detect other targets (such as hydrogen peroxide, glucose, lactic acid, uric acid, creatinine, cholesterol or triglycerides), corresponding biometric recognition elements can be formed on the detection electrode 21 and the calibration electrode 22.

[0072] The exemplary embodiment of the present disclosure also provides an electrochemical biosensor, which includes the above-mentioned detection electrode assembly and detection circuit, the detection circuit is respectively connected to the detection electrode 21 and the calibration electrode 22, and the detection circuit is used to form a reference curve according to the reference identification signal and to form a detection curve according to the detection identification signal.

[0073] In the electrochemical biosensor provided by the embodiments of the present disclosure, during electrochemical detection, the detection electrode 21 generates a detection identification signal in response to the biomaterial to be detected, and the calibration electrode 22 generates a reference identification signal in response to the reference biomaterial. Since the detection electrode 21 and the calibration electrode 22 are located in the same environment and their aging degrees are similar as the electrochemical biosensor is used, the detection result is determined by comparing the reference identification signal and the detection identification signal, which is not affected by the environment and the aging of the detection electrode 21, thereby improving the detection accuracy of the electrochemical biosensor.

[0074] The following describes the detection process of the electrochemical biosensor provided by the embodiment of the present disclosure, taking the content of kanamycin in dairy products as an example:

[0075] Prepare test solutions of varying concentrations: First, add 5 mg of kanamycin to 5 mL of DEPC to create a 1 mg / mL kanamycin stock solution. Then, dilute with DEPC to obtain kanamycin standard test solutions of 50 ng / mL, 100 ng / mL, 150 ng / mL, 250 ng / mL, and 500 ng / mL, respectively. Store these standard test solutions at 4-6°C.

[0076] The prepared Au-Apt aptamer sensor was used as the detection electrode 21 and calibration electrode 22, the Ag / AgCl electrode was used as the reference electrode 23, the carbon electrode was used as the counter electrode 24, and 0.1 M KCl and 5 mM [Fe(CN)6]4- / 3- in PBS with a pH of 7.4 and a concentration of 0.1 mol / L was used as the electrolyte. The DPV curves of the biosensor at different test concentrations were recorded by an electrochemical workstation, as shown in FIG. Figure 5 and Figure 6 shown.

[0077] Depend on Figure 5 and Figure 6 It can be seen that as the concentration of the KANA analyte added to the detection electrode 21 increases, the peak current of the aptamer biosensor shows a downward trend. This phenomenon indicates that specific binding has successfully occurred between the aptamer and its target analyte, kanamycin. In this process, the binding of kanamycin to the aptamer leads to changes in the electrochemical properties of the electrode surface, which in turn affects the generation of current. The reduction in peak current reflects the enhanced interaction between the aptamer and the analyte, providing us with an intuitive way to monitor and quantify the concentration of the target substance. In addition, by comparing the kanamycin test curves of different concentrations with the curves of the calibration electrode 22 separately, it can be clearly observed whether the concentration in the analyte meets the national standard (national standard: the maximum residue limit of kanamycin in milk is 150μg / kg), providing a qualitative and quantitative high-precision, high-reliability detection solution for the dairy field and other biomedical sensing fields, and can maintain stable performance during long-term use.

[0078] Those skilled in the art will readily appreciate other embodiments of the present disclosure after considering the specification and practicing the invention disclosed herein. This application is intended to cover any variations, uses, or adaptations of the present disclosure that follow the general principles of the present disclosure and include common knowledge or customary techniques in the art not disclosed herein. The description and examples are to be considered as exemplary only, with the true scope and spirit of the present disclosure being indicated by the appended claims.

Claims

1. A detection electrode assembly, characterized in that: The detection electrode assembly is used for an electrochemical biosensor, and the detection electrode assembly includes: basal layer; an electrode layer, the electrode layer being disposed on the base layer, the electrode layer comprising a detection electrode and a calibration electrode; wherein, during electrochemical detection, the detection electrode is used to generate a detection identification signal in response to the biomaterial to be detected, and the calibration electrode is used to generate a reference identification signal in response to the reference biomaterial; The electrode layer further includes a counter electrode and a reference electrode. The detection electrode and the counter electrode form a first polarization circuit, the calibration electrode and the counter electrode form a second polarization circuit, and the reference electrode is used to provide a reference potential.

2. The detection electrode assembly according to claim 1, wherein: The base layer has a preset electrode area, the detection electrode and the calibration electrode are arranged in the preset electrode area, and the counter electrode at least partially surrounds the preset electrode area.

3. The detection electrode assembly according to claim 1, wherein: The detection electrode assembly also includes: A conductive layer is provided with a first wire, a second wire, a third wire and a fourth wire, the first wire is connected to the detection electrode, the second wire is connected to the calibration electrode, the third wire is connected to the reference electrode, and the fourth wire is connected to the counter electrode.

4. The detection electrode assembly according to claim 3, wherein: The detection electrode assembly also includes: An insulating layer is provided on the base layer, the insulating layer covers the conductive layer, and a detection opening is provided on the insulating layer, and the orthographic projections of the detection electrode, the calibration electrode, the reference electrode and the counter electrode on the insulating layer are located in the detection opening.

5. The detection electrode assembly according to claim 1, wherein: The detection electrode and the calibration electrode are respectively provided with a biorecognition element, and the biorecognition element is used to identify a preset biomaterial and generate an identification signal.

6. The detection electrode assembly according to claim 5, wherein: The detection electrode is used for detecting kanamycin, and the biological recognition element is a DNA aptamer of kanamycin.

7. An electrochemical biosensor, characterized in that The electrochemical biosensor comprises the detection electrode assembly according to any one of claims 1-6.

8. The electrochemical biosensor according to claim 7, wherein Electrochemical biosensors also include: A detection circuit is connected to the detection electrode and the calibration electrode respectively, and is used to form a reference curve according to the reference identification signal and to form a detection curve according to the detection identification signal.

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