CN215493305U - Electrode, test paper and biosensor for detecting creatinine by electrochemical method - Google Patents
Electrode, test paper and biosensor for detecting creatinine by electrochemical method Download PDFInfo
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Abstract
The utility model provides an electrode, test paper and a biosensor for detecting creatinine by an electrochemical method, wherein the electrode comprises an electrode material base layer and an electronic mediator enhancement layer, and an enzyme composition capable of reacting with creatinine to generate hydrogen peroxide is fixed on the outer surface of the electronic mediator enhancement layer; the test paper comprises an insulating base plate, wherein an electrode group and a circuit are arranged on the surface of the insulating base plate, the electrode group at least comprises a working electrode, a counter electrode and a reference electrode, the working electrode is the electrode of the utility model, and the circuit comprises a loop formed between the working electrode and the counter electrode and between the working electrode and the reference electrode; the sensor comprises an insulating bottom plate, an electrode group, a lead and a sample adding cavity, wherein the electrode group at least comprises a working electrode, a counter electrode and a reference electrode, and the working electrode is the electrode disclosed by the utility model. The test paper and the sensor can be used in various portable and quick small-sized electrochemical detection devices, and can realize quick and convenient clinical detection or home monitoring of blood and urinary creatinine.
Description
Technical Field
The utility model relates to an electrode for analyzing and testing clinical samples, in particular to an electrode for electrochemically detecting creatinine and test paper based on the electrode.
Background
Nowadays, people suffering from kidney diseases, especially Chronic Kidney Disease (CKD), are increasing worldwide. In 2017, global disease, trauma, and risk factor burden (GBD) studies estimated a global prevalence of CKD of 9.1%. The global increase in CKD mortality between 1990 and 2017 for various age groups was 41.5%. With high morbidity and mortality, CKD places a tremendous burden on the global economy. Therefore, it is essential to monitor renal function on a regular basis to avoid the development of renal function injury to an irreversible extent.
Creatinine is the most common indicator of renal function monitoring in clinical work. Creatinine (2-amino-1-methyl-5H-imidazol-4-one) is a chemical waste metabolite of creatine phosphate produced by muscle and protein metabolism. In humans, endogenous creatinine is produced by muscle tissue and exogenous creatinine is primarily from the metabolism of dietary meat. Under normal conditions, creatinine production in our daily lives is relatively stable, and almost all creatinine is filtered by the kidneys and released into the urine. Thus, serum creatinine (SCr) levels will remain within a stable range, approximately 44-106. mu. mol/L. When kidney dysfunction occurs and creatinine excretion capacity is impaired, creatinine concentration will exceed the upper limit of the physiological range. In the early stage, renal patients do not feel discomfort, but an abnormality in the level of SCr draws the attention of the doctor. Therefore, SCr levels are generally considered to be an important biomarker for detecting renal insufficiency.
In the prior art, there are a number of methods for the quantitative determination of creatinine. The traditional methods mainly comprise Jaffe reaction method and enzyme method. Jaffe reaction is fast and economical, but is easily affected by interference chromogens such as ketone, glucose and the like in serum, so that the Jaffe reaction is sometimes inaccurate. Enzymatic methods are therefore considered to be preferred. Indeed, the use of biological enzymes can greatly improve the accuracy and efficiency of detection. Although enzymatic methods place a greater economic burden on patients than the Jaffe reaction, the higher specificity makes the use of enzymatic methods for creatinine detection increasingly the mainstay of clinical testing in hospitals. However, both of these existing creatinine detection methods are not easily implemented for patients who are inconvenient to visit a hospital and who need to be examined regularly for renal function.
To say the daily monitoring of health conditions, POCT-based glucose meters are undoubtedly the most impressive. Unfortunately, no intelligent device such as a blood glucose meter is widely accepted worldwide. A biosensor is an electrochemical detection technique that converts the concentration of a biological material into an electrical signal. Compared with the traditional measuring method, the biosensor has the advantages of short measuring time, simple operation, high sensitivity, good selectivity and the like. Therefore, the electrochemical detection of creatinine using a biosensor is considered to be a promising alternative for creatinine detection.
The prior art discloses methods for electrochemically detecting creatinine and materials used therein. For example, the I-SATA optical and electrical inspection apparatus of yapei-point of care, usa contains a component for performing electrochemical inspection of creatinine, but the apparatus and the component are complicated in mechanism, expensive in cost, not suitable for ordinary patients, and cannot meet the demand for fast and convenient inspection at home. In addition, some existing products have the defects of poor anti-interference capability and low accuracy and sensitivity of detection results.
Due to the complex blood components of the body, the electrochemical detection result of creatinine is easily interfered by other non-target substances in the blood. In order to reduce the detection error caused by such interference, many test materials in the prior art are provided with blood filtration devices for filtering off non-target substances in blood that are likely to cause interference before electrochemical reaction occurs. However, in practice, it is found that the blood filtering device provided in the prior art not only makes the sensor high in cost and complex in process, but also cannot effectively avoid the interference of creatine in blood on creatinine detection.
Therefore, it is necessary to provide an electrochemical electrode and test paper which have simple structure and low cost, and can accurately and quantitatively detect the creatinine in blood, so that the key indexes of kidney diseases can be monitored at home.
SUMMERY OF THE UTILITY MODEL
In view of the above background, the primary object of the present invention is to: the electrode for detecting creatinine by an electrochemical method has the comprehensive advantages of high sensitivity, strong anti-interference performance, stable performance, simple structure, small volume, low manufacturing cost and the like.
Another object of the utility model is: the test paper containing the electrode can be used for quickly and conveniently detecting creatinine clinically or at home.
The above object of the present invention is achieved by the following technical solutions:
in a first aspect, the utility model provides an electrode for detecting creatinine by an electrochemical method, which comprises an electrode material base layer; the outer surface of the electrode material base layer is covered with an electronic mediator enhancement layer; an enzyme reaction layer is fixed on the outer surface of the electronic mediator enhancement layer, and the enzyme reaction layer contains an enzyme composition which can react with creatinine to generate hydrogen peroxide.
In the electrode scheme of the present invention, the material of the electrode material base layer can be any material composition available for redox electrodes, including various compositions containing inert metal materials (such as platinum electrode material, gold electrode material or mercury electrode material) or carbon-containing electrode material, and so on. The material composition may or may not contain an electron mediator component.
In a preferred embodiment of the present invention, the electrode material base layer is a composition mainly composed of a carbon electrode material, and the carbon electrode material may be any one selected from graphite, carbon paste, and glassy carbon; most preferred is carbon paste or graphite.
In a further preferred embodiment of the present invention, the composition mainly comprising carbon electrode material further comprises an electron mediator, wherein the electron mediator may be selected from prussian blue, methylene blue, ferrocene and its derivatives, potassium ferricyanide, and the like; prussian blue is most preferred.
In a second aspect, the utility model provides a test paper for detecting creatinine by an electrochemical method, which comprises an insulating bottom plate, wherein an electrode group and a circuit are arranged on the surface of the insulating bottom plate; the electrode group at least comprises a working electrode, a counter electrode and a reference electrode, wherein the working electrode is the electrode for detecting creatinine by an electrochemical method according to the first aspect of the utility model; the circuit includes a circuit formed between the working electrode and a counter electrode, and between the working electrode and a reference electrode.
The working principle of the test paper used as the electrochemical biosensor is as follows: during detection, after sample is added according to a conventional method, creatinine in a sample can react with an enzyme composition fixed on a working electrode in series, the creatinine is decomposed into creatine by the creatinase, the creatine generates hydrogen peroxide under the action of the creatinase and sarcosine oxidase, the hydrogen peroxide is catalyzed and reduced by an electron mediator in an electron mediator enhancement layer, electrons are transferred to an electrode material base layer of the working electrode to generate reduction current, and the creatine content of the sample can be known according to the relation between a current response value and the concentration of the creatine in the sample. When the test paper is used for detecting creatinine, a detection sample can be in various forms such as whole blood, blood plasma, blood serum, urine and the like.
In the test paper of the present invention, there is no particular limitation on the specific material, specification, and shape of the insulating base plate. The insulating base plate material is preferably insulating paper, plastic, rubber and other materials with a hydrophobic surface; the plastic is further preferably any one of PET, PVC, PE or PP. The specification of the insulating bottom plate can be manufactured into different thicknesses and sizes according to the requirement. The shape of the insulating bottom plate can be any shape which is acceptable in the application scene in actual measurement.
In the test paper of the present invention, the specific shape and specification of each electrode in the electrode group are not particularly limited, and each electrode can be made into various acceptable shapes on the premise that effective electrochemical detection can be achieved, for example, the cross section of the electrode can be rectangular, circular or any other shape.
In the test paper of the present invention, the material of the counter electrode is preferably the same as the electrode material base layer of the working electrode; the reference electrode can be made from a variety of materials that are currently suitable as reference electrodes, with a preferred reference electrode being a silver-silver chloride electrode.
In one embodiment of the present invention, the electrode set may be composed of only one working electrode, one counter electrode and one reference electrode, and the one working electrode is the electrode for electrochemical creatinine detection according to the present invention. The test paper can be used as an electrochemical sensor for independently detecting the creatinine content in a sample, is used in simple and portable creatinine detection equipment, and is suitable for home self-monitoring of nephropathy patients.
In a third aspect, the utility model provides a biosensor for electrochemically detecting creatinine, which comprises an insulating bottom plate, wherein an electrode group, a lead and a sample adding cavity are fixedly connected to the surface of the insulating bottom plate; the sample adding cavity consists of insulating material walls at the periphery and a hollow area in the middle; the electrode group main body is positioned in the range of the hollow area, the electrode group at least comprises a working electrode, a counter electrode and a reference electrode, and the working electrode is the electrode for detecting creatinine by an electrochemical method according to the first aspect of the utility model; the conducting wire comprises metal conducting wires which are respectively connected with the working electrode, the counter electrode and the reference electrode and are used for electrically connecting each electrode with peripheral potential detection equipment.
When the biosensor is used for detecting the creatinine level of blood, a lead of the biosensor is communicated with an external potential detection device, a collected blood sample is dripped into the hollow area of the sample adding cavity in a proper dosage, and the insulating material walls around the hollow area can limit sample liquid drops in the hollow area to be fully contacted with the electrodes. Creatinine in a sample reacts with a compound enzyme on a working electrode to generate hydrogen peroxide, the hydrogen peroxide is catalyzed and reduced by an electronic mediator of an electronic mediator enhancement layer on the working electrode, electrons of an active center of the compound enzyme are transferred to an electrode material base layer, reduction current is generated on the working electrode, potential detection equipment measures potential through a reference electrode, the magnitude of a current response value is related to the blood creatinine level within a certain range, and the corresponding creatinine level is calculated according to a current detection result.
In the biosensor according to the present invention, the specific material, specification and shape of the insulating base plate are not particularly limited. The insulating base plate material is preferably insulating paper, plastic, rubber and other materials with a hydrophobic surface; the plastic is further preferably any one of PET, PVC, PE or PP. The specification of the insulating bottom plate can be manufactured into different thicknesses and sizes according to the requirement. The shape of the insulating bottom plate can be any shape which is acceptable in the application scene in actual measurement.
In the biosensor of the present invention, the specific shape and specification of each electrode in the electrode set are not particularly limited, and each electrode can be made into various acceptable shapes, for example, the cross section of the electrode can be rectangular, circular or any other shape, on the premise that effective electrochemical detection can be achieved. The material of the counter electrode is preferably the same as the electrode material base layer of the working electrode; the reference electrode can be made from a variety of materials that are currently suitable as reference electrodes, with a preferred reference electrode being a silver-silver chloride electrode.
In the biosensor of the present invention, the conductive wire may be a conductive material of various metals, preferably a copper wire.
In the biosensor, the insulating material wall of the sample adding cavity can be made of various existing insulating materials, preferably hot melt adhesive. The preparation method of the electrode and the test paper comprises the following steps:
a method of preparing an electrode according to the first aspect of the utility model, comprising:
1) forming an electrode material base layer from carbon slurry containing an electron mediator and drying;
2) arranging an electronic mediator enhancement layer on the upper surface of the electrode material base layer obtained in the step 1) and drying; for example, the electron mediator enhancement layer can be formed by modifying an electron mediator on the surface of the base layer of the electrode material by an electrodeposition method
3) Immobilizing an enzyme composition on the surface of the electron mediator enhanced layer obtained in the step 2); this can be done, for example, by means of glutaraldehyde crosslinking; the enzyme composition is a composition of creatininase, creatinase and creatininase oxidase; obtaining the enzyme modified electrode, namely the electrode for detecting creatinine by an electrochemical method.
In a preferred method for preparing the electrode of the present invention, 1) the forming of the electrode material base layer with the carbon paste containing the electron mediator is to form the electrode material base layer on an acceptable surface by screen printing the carbon paste based on a pre-designed screen.
The method for preparing the test paper for detecting creatinine by using an electrochemical method according to the second aspect of the present invention comprises:
preparing a working electrode, a counter electrode, a reference electrode and a matched circuit on the surface of an insulating and hydrophobic bottom plate by using carbon paste (such as Prussian blue-doped carbon paste) containing an electron mediator as the working electrode and the counter electrode material and silver-silver chloride paste as the reference electrode material (such as printing by a screen printing mode); applying an electron mediator enhancement layer on the surface of the prepared working electrode (for example, Prussian blue can be modified on the surface of the prepared working electrode by an electrodeposition method); then, an enzyme composition consisting of creatininase, creatinase and sarcosine oxidase is fixed on the surface of the electron mediator enhancement layer (for example, a mixed solution of the enzyme composition and glutaraldehyde is dripped on the surface of the electron mediator enhancement layer and dried at 0-4 ℃; obtaining the test paper capable of detecting creatinine by an electrochemical method.
In practical application, the electrochemical method for detecting creatinine by using the biosensor provided by the utility model comprises the following specific steps:
the biosensor for electrochemically detecting creatinine is used, and a lead of the biosensor is communicated with an external potential detection device; dropping a buffer solution with the pH value of 7.4 into a sample adding cavity of the biosensor, then dropping a to-be-detected blood plasma or whole blood sample into the buffer solution, detecting by using a potential detection device at a working voltage of-0.1V to obtain the intensity of response current, and finally calculating the creatinine concentration in the sample according to a relation curve of the creatinine concentration and a current signal.
Compared with the prior art, the electrode provided by the utility model has the advantages that the internal composition structure is optimized, and the electronic mediator enhancement layer is additionally arranged between the three-enzyme composition for detecting creatinine and the electrode material base, so that the electron transfer efficiency is obviously enhanced, the working potential during detection is reduced to-0.1V, the interference caused by non-target substances such as creatine in creatinine detection is effectively eliminated, and the anti-interference capability of the electrode and the test paper is obviously improved. Meanwhile, the electrode provided by the utility model also has higher sensitivity and repeated measurement stability. In addition, the test paper can be used in various portable and quick small-sized electrochemical detection devices, so that the test paper can realize quick and convenient clinical detection or home monitoring of blood and urinary creatinine.
Drawings
Fig. 1 is a schematic view of the working electrode structure described in example 1.
FIG. 2 is a schematic structural diagram of the creatinine test strip according to example 2.
FIG. 3 is a schematic view of the biosensor according to example 3.
Figure 4 shows the results of a sensitivity study of sensors made with different electrode materials.
Fig. 5 shows the measurement reproducibility of the sensor for detecting creatinine solution of example 3 biosensor.
Fig. 6 shows the correlation between the results of measurement of plasma creatinine concentration using the biosensor of example 3 and the results of measurement of plasma creatinine concentration using the conventional Jaffe method.
Detailed Description
The technical solution of the present invention will be described in detail below by way of examples, but the scope of the present invention is not limited to the examples.
Example 1
An enzyme electrode structure for detecting creatinine by an electrochemical method is shown in figure 1 and comprises an electrode material base layer 1; the outer surface of the electrode material base layer 1 is covered with an electronic mediator enhancement layer 2; an enzyme reaction layer 3 is fixed on the outer surface of the electronic mediator enhancement layer 2, and the enzyme reaction layer 3 contains an enzyme composition which can react with creatinine to generate hydrogen peroxide. The electrode material base layer 1 is made of Prussian blue doped carbon paste; the electronic mediator enhancement layer 2 is a Prussian blue layer modified on the electrode material base layer 1 through electrochemical deposition; the enzyme composition in the enzyme reaction layer 3 is preferably composed of creatininase, creatinase, and sarcosine oxidase.
The method for preparing the enzyme electrode is as follows:
1) coating carbon paste slurry doped with Prussian blue on a PET (polyethylene terephthalate) bottom plate to form a strip shape with the thickness of 0.2mm, and drying to obtain an electrode material base layer 1;
2) continuously scanning Prussian blue on the surface of the electrode material base layer obtained in the step 1) by adopting an electrodeposition method; drying to obtain an electrode with the Prussian blue modified on the surface of the electrode material base layer, wherein the Prussian blue modified on the surface of the electrode material base layer is used as an electronic mediator enhancement layer 2;
3) the enzyme composition is fixed on the surface of the electron mediator enhancement layer 2 obtained in the step 2) by a glutaraldehyde crosslinking method, and an enzyme reaction layer 3 is formed on the surface of the electron mediator enhancement layer 2.
Example 2
The test paper for detecting creatinine through an electrochemical method has a structure shown in fig. 2, and comprises a rectangular PET bottom plate 10, wherein an electrode group and a circuit are arranged on the surface of the PET bottom plate 10; the electrode group consists of an elongated working electrode 20, an elongated counter electrode 30 and an elongated reference electrode 40 which are arranged in parallel, and the structure of the working electrode 20 is the same as that of the electrode in embodiment 1; the counter electrode 30 is made of Prussian blue doped carbon paste; the reference electrode 40 is a silver-silver chloride electrode; the circuit is a metal conductor 50 connecting the electrodes to external test equipment.
Example 3
A biosensor capable of detecting creatinine level of a sample by an electrochemical method is structurally shown in figure 3, and comprises a rectangular PET insulating bottom plate 10 with the specification of 25mm multiplied by 30mm, wherein an electrode group, a lead and a sample adding cavity are fixedly connected to the surface of the insulating bottom plate 10; the sample adding cavity 60 consists of hot melt adhesive walls at the periphery and a hollow area in the middle; the electrode group main body is positioned in the range of the hollow area, the electrode group consists of a working electrode 20, a counter electrode 30 and a reference electrode 40, and the working electrode 20 is the electrode in embodiment 1; the length, width and thickness of each electrode were 20mm, 2mm and 0.2mm respectively. The lead wires comprise copper lead wires 50 which are respectively connected with the working electrode 20, the counter electrode 30 and the reference electrode 40 and are used for electrically connecting each electrode with an external potential detection device. The joint of the copper wire 50 and each electrode is provided with a fixing component 70 for strengthening connection.
The biosensor of the present embodiment can be prepared by:
(1) selecting a rectangular PET material with the specification of 25mm multiplied by 30mm as an insulating base plate 10, and presetting the respective positions of a working electrode 20, a counter electrode 30 and a reference electrode 40 on the insulating base plate 10;
(2) and (2) manufacturing electrodes on the insulating bottom plate 10 in the step (1) according to preset positions, namely, brushing Prussian blue doped carbon paste to obtain a working electrode 20 and a counter electrode 30, and brushing Ag/AgCl ink to obtain a reference electrode 40. The length, width and thickness of each electrode are 20mm, 2mm and 0.2mm respectively, as shown in fig. 3, the middle is the working electrode 20, the left side is the counter electrode 30, and the right side is the reference electrode 40; drying the insulating base plate coated with the electrode in a drying furnace at 75 ℃ for 30 minutes, and cooling at room temperature;
(3) on the cooled insulating bottom plate obtained in the step (2), silver paste is used for connecting copper leads 50 on the ends of the three electrodes on the same side respectively; the attachment portion is then sealed with glue to form a securing assembly 70 to secure the attachment. And then, a raw material wall is built by surrounding the end head which is not connected with the copper wire 50 on the other side of the three electrodes with hot melt adhesive to form a sample adding cavity 60 with a 2mm multiplied by 8mm hollow area in the middle of the surrounding bulge.
(4) Modifying a Prussian blue layer on the surface of the working electrode 20 in the sample adding cavity 60 obtained in the step (3) by an electrodeposition method to serve as an electron mediator enhancement layer;
(5) and (4) fixing an enzyme complex on the surface of the electron mediator enhanced layer obtained in the step (4) in a glutaraldehyde crosslinking mode to obtain the biosensor.
The biosensor prepared in this example was taken out of the refrigerator and incubated in a buffer solution at room temperature for 1 hour before use in the assay. When the biosensor is used for detecting the creatinine level of blood, the copper wire 50 of the biosensor is communicated with an external potential detection device, a buffer solution and a collected blood sample are sequentially dripped into the hollow area of the sample adding cavity 60 in proper dosage, and the insulating material walls around the buffer solution and the collected blood sample can limit liquid drops in the hollow area to be fully contacted with the electrodes. Creatinine in a sample reacts with an enzyme complex on the working electrode 20 to generate hydrogen peroxide, the hydrogen peroxide is catalyzed and reduced by an electron mediator of an electron mediator enhancement layer on the working electrode 20, electrons of an active center of the complex enzyme are transferred to an electrode material base layer, a reduction current is generated on the working electrode 20, a potential detection device measures potential through a reference electrode 40, the magnitude of a current response value is related to the blood creatinine level within a certain range, and therefore the corresponding creatinine level is calculated according to a current detection result.
Comparative example 1
A biosensor useful for electrochemical creatinine detection substantially as described in example 3, except that: the Prussian blue layer is not modified on the working electrode.
Experimental example 1
This experiment investigated the sensitivity of the biosensors of example 3 and comparative example 1. As shown in FIG. 4, the biosensor of comparative example 1 had a detection sensitivity of about 1.4. mu.A/mM; compared with the sensor of the comparative example 1, the sensitivity of the sensor is obviously improved to about 2.2 mua/mM after the PB carbon paste electrode is modified by the Prussian blue electrodeposition layer. The results show that the electron mediator enhancement layer of the present invention can make the sensor show good detection performance at-0.1V potential.
Experimental example 2
To investigate the measurement stability of the biosensor of example 3, the same creatinine solution was measured 74 times in succession. First, 200. mu.L of buffer solution was dropped on the electrode surface, the electrode set was immersed, and after the current baseline of the buffer solution leveled off, 5. mu.L of 41mM creatinine solution was added to the buffer solution, thereby achieving a final concentration of 1mM creatinine. The current response by a 1mM creatinine solution was then recorded. The current response measured at the 1 st time was taken as 100%, and the current responses measured at the subsequent 73 times were quantified based on the response at the 1 st time, in percentage.
The 74 measurements are shown in fig. 5, with the abscissa representing the number of measurements and the ordinate representing the normalized response ratio. It was observed that the current response was stable over the range of up and down 20% (80-120%) based on the current response measured at 1 st, followed by 73 subsequent measurements. It is noteworthy that the sensor still maintained good performance after 74 measurements and that the current response did not decrease but increased compared to the initial signal, probably due to the increased enzyme activity. The result shows that the sensor has good measurement stability when the creatinine is measured continuously.
Experimental example 3
The biosensor of example 3 of the present invention was used to detect creatinine levels in 7 plasma samples, each plasma sample was simultaneously detected by the conventional Jaffe method as a control, and the two detection results and their error rates are shown in table 1 below:
TABLE 1
As shown in FIG. 6, there is also a high correlation between the detection result of the biosensor in example 3 of the present invention and the detection result of the conventional Jaffe method. The biosensor of the utility model is proved to have good accuracy in detecting the creatinine level of blood plasma.
Claims (9)
1. An electrode for detecting creatinine by an electrochemical method comprises an electrode material base layer; the method is characterized in that: the outer surface of the electrode material base layer is covered with an electronic mediator enhancement layer; an enzyme reaction layer is fixed on the outer surface of the electronic mediator enhancement layer, and the enzyme reaction layer contains an enzyme composition which can react with creatinine to generate hydrogen peroxide.
2. The electrode of claim 1, wherein: the cross section or the longitudinal section of the electrode is rectangular or circular.
3. A test paper for detecting creatinine by an electrochemical method, which comprises an insulating bottom plate, and is characterized in that: the surface of the insulating bottom plate is provided with an electrode group and a circuit; the electrode group at least comprises a working electrode, a counter electrode and a reference electrode, wherein the working electrode is the electrode in any one of claims 1-2; the circuit includes a circuit formed between the working electrode and a counter electrode, and between the working electrode and a reference electrode.
4. The test strip of claim 3, wherein: the working electrode, the counter electrode and the reference electrode are in the same shape and are all strip-shaped.
5. The test strip of claim 3, wherein: the working electrode, the counter electrode and the reference electrode are different in shape.
6. A biosensor for electrochemically detecting creatinine comprising an insulating base plate, wherein: the surface of the insulating bottom plate is fixedly connected with an electrode group, a lead and a sample adding cavity; the sample adding cavity consists of insulating material walls at the periphery and a hollow area in the middle; the electrode group main body is positioned in the range of the hollow area, the electrode group at least comprises a working electrode, a counter electrode and a reference electrode, and the working electrode is the electrode in any one of claims 1-2; the conducting wire comprises metal conducting wires which are respectively connected with the working electrode, the counter electrode and the reference electrode and are used for electrically connecting each electrode with peripheral potential detection equipment.
7. The biosensor of claim 6, wherein: the working electrode, the counter electrode and the reference electrode are in the same shape and are all strip-shaped.
8. The biosensor of claim 6, wherein: the working electrode, the counter electrode and the reference electrode are different in shape.
9. The biosensor of claim 6, wherein: and the joints of the metal lead, the working electrode, the counter electrode and the reference electrode are respectively provided with fixing components for strengthening connection.
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