CN211318294U - A Rapid Detection Electrochemical Biosensor - Google Patents

Abstract

The utility model discloses an electrochemical biosensor for rapid detection. An electrode layer is provided with a permeation diffusion layer with permeation and diffusivity, wherein the permeation diffusion layer is covered with at least one enzyme liquid. Drop or adsorb the enzyme solution directly onto the surface of the osmotic diffusion layer, cut the osmotic diffusion layer to a suitable size and fix it on the surface of the electrode, without the need to configure the modification solution, to avoid the thickness of the modification solution to prevent the surface of the electrode from detecting the reaction of the enzyme solution, and the enzyme The problem of wrapping the modification liquid, and the permeable diffusion layer can also filter the blood cells, avoiding the influence of the blood cells on the test results.

Description

A Rapid Detection Electrochemical Biosensor

technical field

The utility model relates to the field of electrochemistry, in particular to an electrochemical biosensor for rapid detection.

Background technique

The following background art is used to help readers understand the present invention, and should not be regarded as prior art.

The main purpose of biosensor research is to develop biosensors with practical application capabilities and their corresponding fabrication techniques. The application of screen printing technology makes it possible to prepare disposable electrochemical biosensor electrodes in batches. The method is simple in process, low in cost, and suitable for industrial production of electrodes. Therefore, screen-printed electrodes based on the principle of electrochemical biosensors are widely used in the field of point-of-care detection (POCT). Physiological fluids such as blood, urine, sweat, and saliva contain many test parameters of interest to clinical laboratories, such as glucose, cholesterol, glycosylated hemoglobin, blood ketones in blood, uric acid in urine, and lactic acid in sweat.

At present, most of the electrode detection principles are to modify the enzyme corresponding to the substrate on the working electrode, and measure the electrical signal through enzymatic reaction. Therefore, the glucose sensor generally adopts the enzymatic method, but the conditions required for the immobilization of the enzyme solution are relatively harsh. , and when the enzyme solution is fixed, first drop the modified solution with a configured concentration on the electrode. The modified solution has the function of diffusing the enzyme solution. After drying, the enzyme solution is added dropwise to make the enzyme solution spread rapidly on the surface of the electrode. This process The thickness of the modification solution in the middle cannot prevent the surface of the electrode from detecting the reaction of the enzyme solution; or directly configure the diffusion modification solution and the enzyme solution together and drop it on the electrode, which is easy to cause the enzyme to be wrapped in the process, and the detection signal will be compared. weak. Moreover, the selection and configuration of the modification solution is also the key. Poor selection can easily affect the reaction and detection results of the enzyme solution, which greatly increases the research and development cost of the electrode test piece.

SUMMARY OF THE INVENTION

The purpose of the present utility model is to provide a rapid detection electrochemical biosensor, the enzyme liquid is directly dropped or adsorbed on the surface of the osmotic diffusion layer, and the osmotic diffusion layer is cut into a suitable size and fixed to the surface of the electrode, without the need to configure a modification liquid, Avoid the thickness of the modification liquid blocking the reaction of the enzyme liquid on the surface of the electrode, and the problem that the enzyme is wrapped by the modification liquid, and the penetration diffusion layer can also filter blood cells, avoiding the influence of blood cells on the test results.

The technical solution adopted by the utility model to solve the technical problem is as follows: an electrochemical biosensor for rapid detection, comprising an electrode layer, the electrode layer is provided with a permeation diffusion layer with permeation and diffusivity, wherein the permeation diffusion layer is covered with at least an enzyme solution. The osmotic diffusion layer adsorbs the enzyme liquid, and the enzyme liquid can quickly diffuse on the osmotic diffusion layer. The sample to be tested first reacts with the enzyme liquid on the osmotic diffusion layer, and the reacted sample penetrates to the surface of the electrode and then interacts with the electrode. Instead of the modification liquid, it plays the role of diffusing the enzyme liquid, and there is no problem that the thickness of the modification liquid blocks the reaction of the electrode surface to detect the enzyme liquid, and the enzyme is wrapped by the modification liquid.

It should be noted that the permeable diffusion layer has permeability that allows the sample to be tested to pass through, and has a diffusion effect. Wherein, the electrode layer has at least two electrodes, respectively a working electrode and a counter electrode, and may also have three electrodes, respectively a working electrode, a counter electrode and a reference electrode. The sample to be tested can be blood, urine or saliva, and the detection parameter can be blood sugar in blood and uric acid in urine. Electrochemical principle of sample detection: When a certain voltage is applied to the sample to be tested after the reaction of the reactant, the generated current will increase with the increase of blood glucose or uric acid concentration in urine. By accurately measuring these weak currents, And according to the relationship between the current value and the blood sugar concentration or uric acid concentration, the corresponding concentration is calculated inversely. Electrochemical detection belongs to the prior art, and its detailed principle and detection process are not repeated here.

Preferably, the permeation diffusion layer is made of fiber. For example non-woven or nylon. Non-woven fabrics or nylon have good penetration and diffusion effects, and can also filter blood cells, avoiding the influence of blood cells on the test results. Preferably, the permeation diffusion layer is filter paper.

Preferably, the pore size of the permeable diffusion layer is 0.1-3um.

Preferably, the electrode has a detection area, and the permeable diffusion layer covers the surface of the detection area. The sample to be tested first reacts with the enzyme solution on the permeable diffusion layer. The reacted sample penetrates into the surface of the electrode and then acts on the electrode. The permeable diffusion layer covers the surface of the detection area. Test results. Among them, the detection area refers to the part that can interact with the enzyme solution.

Preferably, the biosensor comprises a first double-sided adhesive layer and a second double-sided adhesive layer, the first double-sided adhesive layer and the second double-sided adhesive layer are arranged above the electrode layer in sequence, and the first double-sided adhesive layer and the second double-sided adhesive layer are sequentially arranged above the electrode layer. The two double-sided adhesive layers are provided with windows corresponding to each other, the penetration diffusion layer is bonded between the first double-sided adhesive layer and the second double-sided adhesive layer, and the windows correspond to the penetration diffusion layer. The first double-sided adhesive layer and the second double-sided adhesive layer fix the permeable diffusion layer on the electrode layer.

Preferably, the biosensor includes an insulating layer, the electrode layer is located under the insulating layer, the permeation diffusion layer is located above the insulating layer, the insulating layer is provided with a window, and the detection area, the window and the permeation diffusion layer correspond in sequence. The insulating layer can cover the electrodes to prevent them from leaking outside and affecting the detection. It should be noted that the size of the window of the insulating layer is not limited, it needs to be the same size as the area of the detection area or larger than the area of the detection area; the setting shape of the window is not limited, such as square, circular or irregular shape, preferably square ; the window can be closed or have an opening.

Preferably, the biosensor comprises a substrate layer and a hydrophilic membrane layer, the hydrophilic membrane layer is provided with a diversion channel, the diversion channel corresponds to the permeation diffusion layer, the substrate layer, the electrode layer, the insulating layer, the first double-sided The adhesive layer, the permeation diffusion layer, the second double-sided adhesive layer and the hydrophilic film layer are stacked in sequence. The substrate layer is provided with a lead layer, the lead layer includes identification terminals, the lead layer is electrically connected with the electrode layer, and the insulating layer can cover the lead layer to prevent it from leaking outside and affecting detection. The sample to be tested is added from the diversion channel of the hydrophilic membrane layer. The sample to be tested passes through the window of the second double-sided adhesive layer and the osmotic diffusion layer in turn, and reacts with the enzyme liquid on the osmotic diffusion layer. The reacted sample passes through the second double-sided adhesive layer. A window of the double-sided adhesive layer, a window of the insulating layer, and finally the detection area on the electrode layer.

Preferably, the permeation diffusion layer has multiple layers, each permeation diffusion layer is covered with an enzyme solution, and the multiple permeation diffusion layers are stacked in sequence. If a variety of enzyme solutions are coated on a permeable diffusion layer, the reaction of the substrate will be incomplete, and if the thickness is too thick, the activity of the enzyme will be affected, and it may also cause the enzyme to react with the substrate without contacting the electrode. surface, affecting the detection of electrical signals. If there are too many types of interfering substances in the sample to be tested, the modification of various enzymes on the osmotic diffusion layer will have a great influence on the enzymatic activity and the enzymatic reaction. If the enzyme activity becomes low, the removal of interfering substances will be incomplete. When it is multi-layer, each layer is covered with different enzyme liquid, and the sample to be tested reacts with the enzyme liquid permeating the diffusion layer in turn, and the reaction is more sufficient.

Preferably, the electrode layer is modified with a blank enzyme solution, and the osmotic diffusion layer is covered with the enzyme solution. Among them, the enzyme liquid covered on the osmotic diffusion layer may be one or several of them, or all the enzyme liquids used to complete the measurement of the parameters of the sample to be tested. When it is one or several of them, the remaining enzyme liquids Can be modified onto electrodes. Here, the "blank enzyme solution" means that the enzyme solution is not modified on the electrode, and the enzyme solution covered on the osmotic diffusion layer refers to all the enzyme solutions required in the electrochemical detection process, including those that can react with the sample to be tested. The enzyme liquid that generates the detectable electrochemically active product is named in this application as "the enzyme liquid corresponding to the sample to be tested"; the enzyme liquid that can react with the sample to be tested to generate an intermediate product is named in this application as the "intermediate product corresponding to the enzyme liquid". Enzyme liquid"; an enzyme liquid that can react with the interfering substances in the sample to be tested to remove the interfering substances, which is named in this application as "the enzyme liquid corresponding to the interfering substances", etc. For example, when the electrode is modified with a blank enzyme solution, and the permeable diffusion layer is covered with the enzyme corresponding to the sample to be tested, the detection parameters only need one enzyme solution, that is, when the sample to be tested corresponds to the enzyme, the detection can be completed. Preferably, the electrode is modified with a blank enzyme solution, and the permeable diffusion layer is covered with an enzyme solution corresponding to the sample to be tested. Preferably, when the electrode is modified with a blank enzyme solution, and the osmotic diffusion layer is covered with an enzyme solution corresponding to the sample to be tested, the enzyme solution corresponding to the sample to be tested contains 1% wt of glucose oxidase, and the enzyme protective agent trehalose and bovine serum The albumin was 1%wt each, the conductive medium potassium ferricyanide was 20%wt, and the buffer PBS was 67%wt. The electrode modification blank enzyme solution has multiple layers of osmotic diffusion layers. When each osmotic diffusion layer is covered with an enzyme solution, it is respectively covered with the enzyme solution corresponding to the intermediate product, the enzyme solution corresponding to the interfering substance, and the enzyme solution corresponding to the sample to be tested. The samples to be tested are reacted one by one, and the electrochemically active product after the reaction finally penetrates into the electrode for detection.

Preferably, the electrode layer modifies the sample to be tested corresponding to the enzyme solution, and the permeable diffusion layer is covered with the intermediate product corresponding to the enzyme solution. The sample to be tested first reacts with the enzyme solution corresponding to the intermediate product on the permeable diffusion layer to generate an intermediate product, the reacted sample penetrates into the electrode, and the sample to be tested on the electrode reacts with the intermediate product corresponding to the enzyme solution to generate the electrode test strip. Detected electrochemically active products. If all kinds of functional enzyme solutions are immobilized on the electrodes to react directly, there will be great interference between the reaction speeds of each step, and the intermediate products will be incompletely generated, which will affect the next reaction. The product can make the reaction more complete, and each step of the reaction will not interfere with each other. Preferably, the sample to be tested is blood, the detection parameter is cholesterol in the blood, the electrode is modified with cholesterol oxidase, the cholesterol oxidase is the enzyme liquid corresponding to the sample to be tested, the osmotic diffusion layer is covered with cholesterol esterase, and cholesterol esterase is an intermediate The product corresponds to the enzyme solution.

Preferably, the electrode layer modifies the sample to be tested corresponding to the enzyme solution, and the osmotic diffusion layer is covered with interfering substances corresponding to the enzyme solution. The sample to be tested first acts on the osmotic diffusion layer, and the interfering substance on the osmotic diffusion layer reacts with the interfering substance of the corresponding enzyme solution and the sample to be tested, and the reacted sample penetrates into the electrode and reacts with the corresponding enzyme solution of the sample to be tested to form an electrode test strip Detectable electrochemically active products. Some samples to be tested contain a variety of interfering substances. At this time, multiple layers of osmotic diffusion layers can be set up in sequence. Each osmotic diffusion layer corresponds to an interfering substance corresponding to an enzyme solution. The interfering substances in the layer are reacted with the enzyme solution one by one to completely remove the interfering substances. Preferably, the sample to be tested is blood, the detection parameter is uric acid in the blood, the electrode is modified with uricase, the uricase is the enzyme solution corresponding to the sample to be tested, the osmotic diffusion layer is covered with ascorbate oxidase, and the ascorbate oxidase is the interference The substance corresponds to the enzyme solution.

Preferably, the modified interfering substance in the electrode layer corresponds to the enzyme solution, and the permeable diffusion layer is covered with the sample to be tested corresponding to the enzyme solution. After the sample to be tested acts on the permeable diffusion layer, the corresponding enzyme solution of the sample to be tested reacts with the sample to be tested to generate an electrochemically active product that can be detected by the electrode test strip. Interfering substances in the sample.

Preferably, the treatment process of the osmotic diffusion layer is as follows: dropping the enzyme solution on the osmotic diffusion layer or soaking the osmotic diffusion layer in the enzyme solution, and then drying at 37° C. for 10 minutes.

The beneficial effects of the present utility model:

1. By adding a permeable diffusion layer covered with enzyme liquid to replace the modification liquid, it plays a role in the diffusion of the enzyme liquid, without considering the diffusion of the enzyme liquid on the electrode surface, avoiding the thickness of the modification liquid to block the reaction of the electrode surface to detect the enzyme liquid , and the problem that the enzyme is wrapped in the modification liquid, the detection sensitivity is high, the linear range is wide, the accuracy of the electrode is improved, the result can be obtained in about 5S, and the reading operation is simple, which is of great significance for the use of electrochemical sensors.

2. There is no need to consider a variety of enzyme liquid modification conditions, which reduces the production cost and simplifies the production conditions.

3. The osmotic diffusion layer is made of fiber material, which plays the role of filtering blood cells and avoids the influence of blood cells on the test results.

4. Selectively modifying the enzyme solution on the electrode or the osmotic diffusion layer to carry out the step-by-step reaction can make the enzymatic reaction of the sample to be tested more sufficient, the final substrate is more uniform, and the detection result is more accurate.

Description of drawings

Figure 1 is a schematic structural diagram of the utility model.

2 is a time-current graph detected on an electrode provided with a permeation diffusion layer in Example 1. FIG.

FIG. 3 shows the fitting linearity of the time-current graph detected on the electrode provided with the permeation diffusion layer in Example 1. FIG.

Labels in the figure: substrate layer 1, electrode layer 2, insulating layer 3, first double-sided adhesive layer 4, permeable diffusion layer 5, second double-sided adhesive layer 6, hydrophilic film layer 7, counter electrode 31, working electrode 32, the reference electrode 33, the window 31 on the insulating layer, the window 41 on the first double-sided adhesive layer, the window 61 on the second double-sided adhesive layer, and the diversion channel 71.

Detailed ways

The structures involved in the present invention or the used technical terms are further described below. These descriptions only illustrate how the present invention is implemented by way of examples, and do not constitute any limitation to the present invention.

The present utility model will be further described below with reference to the accompanying drawings and specific embodiments. In the description of the present invention, it should be understood that the orientations or positional relationships indicated by the terms "upper", "lower", "front", "rear", "left" and "right" are based on those shown in the accompanying drawings. The orientation or positional relationship is only for the convenience of describing the present utility model and simplifying the description, rather than indicating or implying that the indicated position or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as the present invention. limit. Furthermore, the terms "first," "second," etc. are used for descriptive purposes only and should not be construed to indicate or imply relative importance.

As shown in FIG. 1, an electrochemical biosensor for rapid detection includes an electrode layer 2, characterized in that the electrode layer 2 is provided with a permeable diffusion layer 5, wherein the permeable diffusion layer 5 is covered with at least one enzyme solution. The osmotic diffusion layer 5 adsorbs the enzyme liquid, and the enzyme liquid can quickly diffuse on the osmotic diffusion layer 5. The sample to be tested first reacts with the enzyme liquid on the osmotic diffusion layer 5, and the reacted sample penetrates to the surface of the electrode layer 2 and then interacts with the electrode. By adding the osmotic diffusion layer 5 to replace the modification liquid, it plays the role of diffusing the enzyme liquid, avoiding the problem that the thickness of the modification liquid blocks the reaction of detecting the enzyme liquid on the surface of the electrode, and the enzyme is wrapped by the modification liquid.

It should be noted that the permeable diffusion layer 5 has permeability allowing the passage of the sample to be tested and the enzyme liquid, and has the function of diffusing the enzyme liquid. The electrode layer 2 has at least two electrodes, respectively a working electrode and a counter electrode, and may also have three electrodes, which are a working electrode 22 , a counter electrode 21 and a reference electrode 23 respectively, see FIG. 1 . The sample to be tested can be blood, urine or saliva, and the detection parameter can be blood sugar in blood and uric acid in urine. Electrochemical principle of sample detection: When a certain voltage is applied to the sample to be tested after the reaction of the reactant, the generated current will increase with the increase of blood glucose or uric acid concentration in urine. By accurately measuring these weak currents, And according to the relationship between the current value and the blood sugar concentration or uric acid concentration, the corresponding concentration is calculated inversely. Electrochemical detection belongs to the prior art, and its detailed principle and detection process are not repeated here.

The permeation diffusion layer 5 is made of fiber material. For example non-woven or nylon. Non-woven fabrics or nylon have good penetration and diffusion effects, and can also filter blood cells, avoiding the influence of blood cells on the test results.

The permeation diffusion layer 5 is filter paper.

The pore size of the permeable diffusion layer 5 is 0.1-3um.

The electrode layer 2 has a detection area, and the permeable diffusion layer 5 covers the surface of the detection area. The sample to be tested first reacts with the enzyme liquid on the permeable diffusion layer 5, the reacted sample penetrates to the surface of the electrode and then interacts with the electrode. Get test results. Among them, the detection area refers to the part where the electrode can interact with the enzyme solution.

The biosensor includes a first double-sided adhesive layer 4 and a second double-sided adhesive layer 6, the first double-sided adhesive layer 4 and the second double-sided adhesive layer 6 are sequentially arranged above the electrode layer 2, and the first double-sided adhesive layer is 4 and the second double-sided adhesive layer 6 both have windows corresponding to each other. The first double-sided adhesive layer 4 and the second double-sided adhesive layer 6 fix the permeable diffusion layer on the electrode layer 2 .

The biosensor includes an insulating layer 3 , an electrode layer 2 is located below the insulating layer 3 , and a permeable diffusion layer 5 is located above the insulating layer 3 , the insulating layer 3 has a window 31 , and the detection area and the window 31 correspond to the permeable diffusion layer 5 in sequence. The insulating layer 3 can cover the electrodes to prevent them from leaking outside and affecting detection. It should be noted that the size of the window 31 of the insulating layer 3 is not limited, it needs to be the same size as the area of the detection area or larger than the area of the detection area; the shape of the window 31 is not limited, such as square, circular or irregular shape , preferably square; the window 31 can be closed or have an opening.

The biosensor includes a substrate layer 1 and a hydrophilic membrane layer 7. The hydrophilic membrane layer 7 is provided with a diversion channel 71, and the diversion channel 71 corresponds to the permeation diffusion layer 5. The substrate layer 1, the electrode layer 2, the insulating layer 3. The first double-sided adhesive layer 4, the permeable diffusion layer 5, the second double-sided adhesive layer 6 and the hydrophilic film layer 7 are stacked in sequence. The substrate layer 1 is provided with a wire layer 11, the wire layer 11 includes identification terminals, the wire layer 11 is electrically connected to the electrode layer 2, and the insulating layer 3 can cover the wire layer 11 to prevent it from leaking outside and affecting detection. The sample to be tested is added from the diversion channel 71 of the hydrophilic membrane layer 7, and the sample to be tested passes through the window 61 of the second double-sided adhesive layer 6 and the osmotic diffusion layer 5 in sequence, and reacts with the enzyme liquid on the osmotic diffusion layer 5. The latter sample passes through the window 41 of the first double-sided adhesive layer 4 and the window 31 of the insulating layer 3 , and finally acts on the detection area on the electrode layer 2 .

The permeation diffusion layer 5 has multiple layers, each permeation diffusion layer 5 is covered with an enzyme solution, and the multiple permeation diffusion layers are stacked in sequence. If a variety of enzyme liquids are coated on one layer of permeable diffusion layer 5, the reaction of the substrate will be incomplete, and if the thickness is too thick, the activity of the enzyme will be affected, and the enzyme may not be in contact with the substrate after the reaction. The electrode surface affects the detection of electrical signals. If there are too many types of interfering substances in the sample to be tested, the modification of various enzymes on the osmotic diffusion layer will have a great influence on the enzymatic activity and the enzymatic reaction. When it is multi-layer, each layer is covered with different enzyme liquid, and the sample to be tested reacts with the enzyme liquid permeating the diffusion layer in turn, and the reaction is more sufficient.

As an embodiment, the electrode layer 2 is modified with a blank enzyme solution, and the permeable diffusion layer 5 is covered with the enzyme solution. Among them, the enzyme liquid covered on the osmotic diffusion layer may be one or several of them, or all the enzyme liquids used to complete the measurement of the parameters of the sample to be tested. When it is one or several of them, the remaining enzyme liquids Can be modified onto electrodes. Here, the "blank enzyme solution" means that the enzyme solution is not modified on the electrode, and the enzyme solution covered on the osmotic diffusion layer 5 refers to all the enzyme solution required in the electrochemical detection process, including the enzyme solution that is compatible with the sample to be tested. The enzyme liquid that reacts to generate a detectable electrochemically active product is named in this application as "the enzyme liquid corresponding to the sample to be tested"; the enzyme liquid that can react with the sample to be tested to generate an intermediate product is named in this application as "intermediate product" Corresponding enzyme solution"; the enzyme solution that can react with the interfering substance in the sample to be tested to remove the interfering substance is named in this application as "the enzyme solution corresponding to the interfering substance", etc. For example, in one case: when the electrode is modified with a blank enzyme solution, and the permeable diffusion layer is covered with the enzyme corresponding to the sample to be tested, the detection parameters only need one enzyme solution, that is, when the sample to be tested corresponds to the enzyme, the detection can be completed. It is as follows: the content of glucose oxidase contained in the corresponding enzyme solution of the sample to be tested is 1%wt, the enzyme protective agent trehalose and bovine serum albumin are each 1%wt, and the content of the conductive medium potassium ferricyanide is 20%wt, The content of the buffer PBS was 67% wt, and the glucose oxidase was the enzyme corresponding to the sample to be tested modified on the osmotic diffusion layer. Another case is: the electrode modified blank enzyme solution has multiple layers of osmotic diffusion layers, and each osmotic diffusion layer is covered with an enzyme solution, which is respectively covered with the enzyme solution corresponding to the intermediate product, the enzyme solution corresponding to the interfering substance, and the enzyme solution corresponding to the sample to be tested. In this way, the samples to be tested are added to react one by one, and finally the electrochemically active product after the reaction penetrates into the electrode for detection, the reaction is fully and thoroughly, and the detection result is accurate.

As an embodiment, the electrode layer 2 modifies the enzyme liquid corresponding to the sample to be tested, and the permeable diffusion layer 5 is covered with an intermediate product corresponding to the enzyme liquid. The sample to be tested first reacts with the enzyme solution corresponding to the intermediate product on the permeable diffusion layer to generate an intermediate product, the reacted sample penetrates into the electrode, and the sample to be tested on the electrode reacts with the intermediate product corresponding to the enzyme solution to generate the electrode test strip. Detected electrochemically active products. If all kinds of functional enzyme solutions are immobilized on the electrodes to react directly, there will be great interference between the reaction speeds of each step, and the intermediate products will be incompletely generated, which will affect the next reaction. The product can make the reaction more complete, and each step of the reaction will not interfere with each other.

As an embodiment, the sample to be tested is blood, the detection parameter is cholesterol in the blood, the electrode is modified with cholesterol oxidase, the cholesterol oxidase is the enzyme liquid corresponding to the sample to be tested, the osmotic diffusion layer is covered with cholesterol esterase, cholesterol ester The enzyme is the intermediate product corresponding to the enzyme liquid.

As an embodiment, the electrode layer 2 modifies the enzyme liquid corresponding to the sample to be tested, and the osmotic diffusion layer 5 is covered with an interfering substance corresponding to the enzyme liquid. The sample to be tested first acts on the osmotic diffusion layer, and the interfering substance on the osmotic diffusion layer reacts with the interfering substance of the corresponding enzyme solution and the sample to be tested, and the reacted sample penetrates into the electrode and reacts with the corresponding enzyme solution of the sample to be tested to form an electrode test strip Detectable electrochemically active products. Some samples to be tested contain a variety of interfering substances. At this time, multiple layers of osmotic diffusion layers stacked in sequence can be provided. Each osmotic diffusion layer 5 corresponds to an interfering substance corresponding to an enzyme solution. The interfering substances correspond to the enzyme solution to react the interfering substances in the sample to be tested one by one to completely remove the interfering substances. Preferably, the sample to be tested is blood, the detection parameter is uric acid in the blood, the electrode is modified with uricase, the uricase is the enzyme solution corresponding to the sample to be tested, the osmotic diffusion layer is covered with ascorbate oxidase, and the ascorbate oxidase is the interference The substance corresponds to the enzyme solution.

As an embodiment, the electrode layer 2 modifies the interfering substance corresponding to the enzyme liquid, and the permeable diffusion layer 5 is covered with the enzyme liquid corresponding to the sample to be tested. After the sample to be tested acts on the permeable diffusion layer, the corresponding enzyme solution of the sample to be tested reacts with the sample to be tested to generate an electrochemically active product that can be detected by the electrode test strip. Interfering substances in the sample.

The treatment process of the osmotic diffusion layer 5 is as follows: dropping an enzyme solution on the osmotic diffusion layer 5 or soaking the osmotic diffusion layer 5 in the enzyme solution, and then drying at 37° C. for 10 minutes.

Example 1

The osmotic diffusion layer 5 is covered with glucose oxidase liquid, the sample to be tested is blood, and the blood glucose concentration in the blood is detected. At room temperature, the glucose biosensor for rapid detection of whole blood glucose was tested by the time-amperometry method for whole blood with different blood glucose concentrations of 0-20 mmol/L at a voltage of 0.55V, and the detection time was selected from the time-current curve as 3s respectively. Multiple detection points of 4s, 4s and 5s are marked on the blood glucose concentration-current graph, and the blood glucose concentration-current detection points with detection time of 3s, 4s and 5s are linearly fitted to obtain the blood glucose concentration-current linear fitting graph . The square of the correlation coefficient (R ² ) in the figure indicates how closely the current is related to the blood glucose concentration. When R ² is closer to 1, it means that the linear fitting is more accurate and the detection accuracy of the blood glucose sensor is higher; on the contrary, the more R ² When it is close to 0, it means that the linear fitting is less accurate and the detection accuracy of the blood glucose sensor is lower. The squared correlation coefficient of the test value of the glucose biosensor for blood glucose is greater than 0.99, with good accuracy.

The invention shown and described herein may be practiced in the absence of any element, limitation, specifically disclosed herein. The terms and expressions employed are to be used as terms of description rather than limitation and it is not intended that the use of these terms and expressions exclude any equivalents of the features shown and described, or parts thereof, and it should be recognized that each All modifications are possible within the scope of the present invention. It should therefore be understood that although the present invention has been specifically disclosed by various embodiments and optional features, modifications and variations of the concepts described herein may be employed by those of ordinary skill in the art and are considered to fall within the scope of within the scope of the present invention as defined in the appended claims.

The contents of articles, patents, patent applications, and all other documents and electronically available information described or recorded herein are hereby incorporated by reference in their entirety to the same extent as if each individual publication was specifically As if indicated separately for reference. Applicants reserve the right to incorporate into this application any and all materials and information from any such articles, patents, patent applications or other documents.

Claims (9)

1. The electrochemical biosensor for fast detection comprises an electrode layer, and is characterized in that the electrode layer is provided with a permeation diffusion layer with permeation and diffusion properties, wherein the permeation diffusion layer is covered with at least one enzyme solution.

2. The electrochemical biosensor for rapid detection according to claim 1, wherein the pervaporation layer is made of fiber.

3. The electrochemical biosensor for rapid detection according to claim 2, wherein the pervaporation layer is filter paper.

4. The electrochemical biosensor for rapid detection according to claim 3, wherein the pore size of the pervaporation layer is 0.1-3 um.

5. The electrochemical biosensor for rapid detection according to claim 1, wherein the electrode layer has a detection region, and the pervaporation layer covers the surface of the detection region.

6. The electrochemical biosensor for rapid detection according to claim 5, wherein the biosensor comprises a first double-sided adhesive layer and a second double-sided adhesive layer, the first double-sided adhesive layer and the second double-sided adhesive layer are respectively provided with a window corresponding to each other, the pervaporation layer is located between the first double-sided adhesive layer and the second double-sided adhesive layer, and the window corresponds to the pervaporation layer.

7. The electrochemical biosensor for rapid detection according to claim 6, wherein the biosensor comprises an insulating layer, an electrode layer is disposed under the insulating layer, the insulating layer has a window corresponding to the detection region, a pervaporation layer is disposed over the window, and the electrode layer, the insulating layer, the first double-sided adhesive layer, the pervaporation layer and the second double-sided adhesive layer are sequentially stacked.

8. The electrochemical biosensor for rapid detection according to claim 7, wherein the pervaporation layer has a plurality of layers, each layer of the pervaporation layer is coated with an enzyme solution, and the plurality of layers of the pervaporation layer are sequentially stacked.

9. The electrochemical biosensor for rapid detection according to claim 8, wherein the electrode layer modifies blank enzyme solution, and the permeation diffusion layer is covered with enzyme solution; or modifying the enzyme solution corresponding to the sample to be detected by the electrode layer, and covering the intermediate product corresponding to the enzyme solution by the permeation diffusion layer; or the electrode layer modifies enzyme liquid corresponding to the sample to be detected, and the permeation diffusion layer is covered with enzyme liquid corresponding to the interferent; or the electrode layer modifies the interferent corresponding enzyme liquid, and the permeation diffusion layer is covered with the enzyme liquid corresponding to the sample to be detected.

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