CN1349096A - Electrochemical electrode test piece and manufacturing method thereof - Google Patents

Abstract

An electrochemical electrode test piece and its manufacturing method, which utilizes the specific biochemical or chemical oxidation-reduction reaction between the redox active substance and the specific substance in the sample to generate electron transfer, and the electrochemical electrode test piece matched with the sensor device to detect the concentration of the substance to be detected, the electrode test piece at least includes the electrode part for transferring the electron transfer effect generated in the specific biochemical or chemical oxidation-reduction reaction, the reaction film layer, the sample bearing space above the biological reaction layer and the upper cover protection plate. Wherein the sample bearing space has three openings.

Description

Electrochemical electrode test piece and manufacturing method thereof

The present invention relates to an electrochemical electrode test piece and its manufacturing method, which relates to a specific biochemical or chemical oxidation-reduction reaction by contacting a reaction film of an active material with redox characteristics with a sample, The electrochemical detection principle that produces electron transfer has a low sample volume, fast and accurate disposable electrochemical electrode test piece.

As we all know, electrode test strips that detect the concentration of specific substances in liquids based on electrochemical principles have been used for a long time. At present, there are many blood glucose test strips for self-testing at home on the market, which apply this technology. However, due to the unsatisfactory design of the reaction area for dropping the sample, the method of dropping the sample and the demand for the sample often cause problems, resulting in contamination of the reaction area and insufficient sample volume, resulting in deviations in measurement. For example, too little sample does not completely cover the reaction area, or the time taken for the sample to completely fill the reaction area is too long, resulting in poor reaction time, or fingertips touching the surface of the reaction area contaminates the test piece, resulting in errors in test results. Especially for the elderly or infants, the collection of blood specimens with blood collection needles cannot reach an appropriate amount, thus often causing errors in measurement and wasting test pieces. It is the trend of the times to drop specimens and design electrode strips with less demand for specimens. At present, the ideal goal is to use siphon test strips with a blood volume of less than 5uL.

Currently commercially available blood glucose electrode test strips. As shown in Figure 1, it includes an electrical insulation substrate 101, on which there is a bipolar electrode system 102, the second layer of electrical insulation layer 105 covers the electrical insulation substrate 101, and exposes the electrode surface of the reaction zone 104 and Connect the cathode and anode surfaces of the detector. The reaction area 104 is covered with reaction reagents, and then covered with the net 106 after drying, and finally pasted with adhesive tape 107 to fix the net 106 and protect the reaction area. The net 106 is completely located above the reaction area. In addition, the adhesive tape on both sides of the net 106 has two cracks 108, 109 for exhausting air and reducing air bubbles.

However, the sample demand for this electrode test piece can only be reduced to 9ul, and the exhaust effect of the two cracks in the adhesive tape is extremely poor.

Currently another commercially available blood glucose electrode test strip, as shown in Figure 2, the electrode test strip includes an electrode system 204, 205 on a substrate 201, and a specimen carrying space above the electrodes 204, 205 in the reaction zone 207, wherein the sample carrying space 207 has a sample suction port 208 and a gas discharge port 209. The production of the reaction layer of this blood glucose electrode test strip is to apply carboxymethyl cellulose CMC (carboxymethyl cellulose) aqueous solution on the surface of the electrodes 204, 205 in the reaction area in sequence, and carry out the first drying to form a layer of hydrophilic Permanent polymer CMC layer. Continue to apply glucose oxidase (GOD) solution on it. After drying for the second time, a hydrophilic polymer (polyvinyl azapentane PVP) solution is sprayed on it. After the third drying, a protective film is formed, and then an organic suspension solution containing electronic mediators is sprayed on the PVP protective film. Afterwards, it is dried for the fourth time to complete the biochemical reaction area. Finally, the insulating layer 206 and the upper cover protection plate 210 forming the specimen carrying space are covered on the substrate 201 to complete the blood glucose electrode test piece.

The above-mentioned blood glucose electrode test piece, the biological reaction layer is divided into four steps to complete, that is, the formation of the CMC layer, the GOD layer, the hydrophilic PVP layer and the electronic medium layer. Each layer needs to be dried first, and the process steps are complicated. Moreover, after the sample is inhaled from the suction port, it takes a while to completely fill the sample carrying space and activate the electrode detection side. There is a time difference in the measurement, resulting in measurement error.

The main purpose of the present invention is to provide an electrochemical sheet-shaped electrode test piece with three sample inlets, which can arbitrarily suck the sample through one of the inlets, and the other two openings are gas outlets, providing a more convenient Operation method.

The second object of the present invention is to provide an electrochemical sheet electrode test piece with different designs, which has a sample inlet and two gas outlets. The time difference for the sample to fill the reaction space can be shortened to reduce the error of the detection result.

The third object of the present invention is to provide an external design of the electrode test strip, in which the two ends of the reagent reaction area protrude so that the user can easily touch and carry the sample.

The fourth object of the present invention is to provide an electrochemical sheet-shaped electrode test strip with a sample requirement of less than 5uL.

The fifth object of the present invention is to provide an electrochemical sheet-shaped electrode test strip with a fixed sample requirement, so as to reduce detection errors caused by different sample quantities.

The sixth object of the present invention is to provide an electrochemical sheet-shaped electrode test piece that can be easily manufactured, simplifies the manufacturing process of the reaction film layer, and can be manufactured quickly and in large quantities.

The object of the present invention is achieved in that the electrochemical bioelectrode test strip of the present invention comprises: an electrically insulating flat substrate, and a conductive film positioned on the upper surface of the flat electrically insulating substrate to form two anodes that are not connected to each other. part and the cathode part. A reaction space area, covering the same side surface of the conductive film on the flat electrical insulating substrate with a second layer of padding insulating flat plate to form a reaction area of a groove channel, so that the anode part of the conductive film is not raised by the pad One end of the bare part covered by the insulating flat forms a reference electrode and the other end forms an anode joint, and one end of the exposed part of the cathode part of the conductive film that is not covered by the raised insulating layer forms a working electrode and the other end forms a cathode joint; And a reaction film layer, which is composed of a solution containing at least one active substance with redox properties, covering the reaction area of the aforementioned groove channel, wherein the active substance includes oxidoreductase, redox protein and redox-containing substances, etc.; the solution still contains polymers, electronic mediators and buffer solutions.

The main advantage of the present invention is to use the electrochemical electrode test strips that have specific biochemical or chemical redox reactions to produce electron transfer between active substances with redox characteristics and specific analytes in the sample, which cooperate with the sensor device It can be used to measure the content of the analyte in the sample very specifically and sensitively. Since the detection method of the electrode test piece in the present invention adopts the electrochemical principle, the quantitative determination of the analyte in the sample can be directly carried out. The upper cover protection plate is an electric insulation plate with a hole, which is used to fix the space of the reaction zone and prevent foreign matters from entering the reaction zone.

Below in conjunction with preferred embodiment and accompanying drawing, describe in detail as follows:

1 is a schematic diagram of the first commercially available blood glucose electrode test strip;

2 is a schematic diagram of another commercially available blood glucose electrode test strip;

Fig. 3 is the top view of the electrode test piece of embodiment 1 of the present invention;

Fig. 4 is the front view of Fig. 3;

Fig. 5 is a side view of Fig. 3;

6 is a schematic diagram of the manufacturing process step (1) of the electrode test piece in Example 2 of the present invention;

7 is a schematic diagram of the manufacturing process step (2) of the electrode test piece of Example 2 of the present invention;

8 is a schematic diagram of the manufacturing process step (3) of the electrode test piece in Example 2 of the present invention;

9 is a schematic diagram of the manufacturing process step (four) of the electrode test piece of Example 2 of the present invention;

Fig. 10 is a graph comparing the measurement results of the present invention with those of the biochemical analyzer.

Example 1

Referring to Fig. 3-Fig. 5, the electrochemical electrode test piece of the present invention is a rectangular piece. Its structure roughly includes a flat electrical insulating substrate 301, a conductive film 302 located on the surface of the electrical insulating substrate 301, a raised insulating flat plate 311 partially covering the conductive film 302 to form a groove channel, and a reaction that reacts with the specimen. Membrane 303, and an upper cover protection plate 306 with a hole.

The electrical insulation plate substrate 301 in the present invention preferably has a flat surface, and has electrical insulation properties and heat resistance of heat treatment above 60 degrees Celsius, so as to facilitate the heating, drying and curing treatment of the conductive film 302 and the reaction layer 303 , suitable materials such as polyethylene terephthalate board (PET), polyvinyl chloride board (PVC), polycarbonate board (PC), glass fiber board (FR-4), glass board, bakelite board, ceramic board (CEM-1), polyester board (polyester supbone) and the like.

The conductive film 302 includes at least two separated and disconnected anode parts and cathode parts, which are used to connect the sensor device; the cathode part is partially covered with a padding insulating plate 311, and the exposed two ends are respectively the working electrode and the cathode connector, and The working electrode of the cathode part is then covered with a reaction film 303 to detect the electrical effect induced by the sample during the electrochemical reaction, and the cathode connector is used to connect the sensing device. The anode part is also partly covered with a raised electrical insulating flat plate 311, and the exposed two ends are respectively the reference electrode and the anode joint, and the reference electrode of the anode part is covered with a reaction film 303 to cooperate with the working electrode to have an electrical effect on the specimen. detection. The anode connector is used to connect the sensing device.

In the electrochemical electrode test strip according to the present invention, the insulating flat plate 311 is covered on the same side surface of the electrically insulating substrate 301 without covering the anode joint, cathode joint, working electrode and reference electrode. And form a groove channel. A suitable thickness of the raised insulating plate 311 is 0.2 mm or more. The groove channel not covering the raised insulating plate 311 includes two parts, a working electrode and a reference electrode, forming a reaction area, which is covered by the reaction film 303 to accommodate the sample.

The reaction film 303 is an active substance with redox properties, such as redox enzymes, redox proteins, redox compounds, etc.; the mixture solution includes polymers, electronic mediators, non-ionic surfactants and buffer solutions. Among them, high molecular polymers are used to immobilize redox active substances and electronic mediators. This macromolecular polymer accounts for 1-4% weight percent in the reaction film reagent, can be selected from chitin chitin, albumin albumin, polyvinyl azapentacyclone PVP (PolyvinylPyrrolidone), polyethylene glycol PEG (Polyethylene G1vcol ), gelatin, dextran, polyvinyl alcohol PVA (polyvinyl alcohol), polyethyl acetate PVAc (polyvinyl acetate), methylcellulose methylcellulose, carboxymethylcellulose carboxymethylcellulose.

Another component of the reactive membrane 303 becomes the electron mediator. It is used to receive or provide the electron transfer generated during the specific biochemical or chemical redox reaction between the redox active substance and the analyte in the specimen. The voltage, resistance or current change of the electrochemical reaction at this time can be conducted from the working electrode and the reference electrode in contact with the conductive film 302 and the reaction film 303 to the cathode joint and the anode joint at the other end of the conductive film. When this electrochemical electrode test piece is connected with a sensor, the sensor can give the electrode test piece an external voltage through a voltage output device, and a signal receiving device, and the voltage, resistance or The current change is received, and the signal is converted into the concentration of the analyte to be displayed by a display device.

In the specific embodiment of the present invention, potassium ferricyanide is used as the electron mediator, and the electron mediator of the reaction membrane component accounts for 1-10% by weight of the reaction membrane composition formula.

Another component of the reaction film 303 is a non-ionic surfactant, which is used to disperse and cover the reaction reagent to the hydrophobic reaction area. The surfactant accounts for less than 0.3% by weight of the reagent composition formula. Suitable nonionic surfactants include Triton X-100, lecithine; phosphatidylcholine; oleic acid; cyclodextrin and Polyoxyethylene, glycerine, fatty acidester.

Another component of the reaction membrane 303 is a buffer solution, which can be composed of borate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, trisformaldehyde aminomethane (Tris) or citric acid and deionized water. 83.7-98% by weight. This buffer solution is used to maintain optimal activity of the overall reaction when in contact with the specimen.

In the electrode test piece according to the present invention, a hole is covered with a protective plate 306, which is used to protect the reaction membrane from contamination, and to limit the quantitative entry of the sample into the reaction area. At the same time, three openings are formed, one of which can be selected arbitrarily. The entrance is the sample introduction port, and the other two openings are the gas discharge ports. On the one hand, it can shorten the time difference for the sample to fill the reaction zone space. In order to reduce the detection error, on the other hand, it provides more convenient operation for users.

Another feature of the present invention is that the reaction membrane reagent is a single mixed solution, which only needs to be produced and coated in the reaction area in one process, and does not require two-stage production procedures of redox active substances and electronic media. The conditions are tightly controlled, so it is suitable for mass production and reduces costs.

The present invention also provides a method for manufacturing an electrochemical electrode test piece, comprising the following steps:

(1) Covering a conductive film on one surface of an electrically insulating plate-shaped substrate, the conductive film forms two mutually disconnected anode parts and cathode parts;

(2) Covering a raised insulating plate to partially cover the conductive film to form a groove channel, one end of the exposed part of the anode part of the conductive film not covered by the raised insulating plate forms at least one reference electrode and the other end forms an anode Connector, one end of the exposed part of the cathode part of the conductive film that is not covered by the electrically insulating flat plate forms at least one working electrode, and forms a cathode joint with the other end;

(3) The reaction film is covered in the reaction area of the groove channel in a one-time program coating method. The reaction film is composed of an active substance with redox properties and a mixed solution, wherein the mixed solution includes a polymer containing material, electronic medium, non-ionic surfactant and buffer solution;

(4) Cover the upper cover protection plate with a hole on the insulating plate, and form three openings in the reaction area.

Example 2

Referring to Fig. 6-Fig. 9, according to the method for manufacturing the electrochemical electrode test piece of the present invention, at first, on any flat surface of a plate-like electrical insulating substrate, print out an anode that at least one layer is independent and not connected with each other by screen printing and a cathode conductive film 302, as shown in FIG. 6 . The material of the conductive film can be carbon glue, silver glue, gold glue, copper glue, carbon-silver mixed glue, carbon-palladium mixed glue, carbon-platinum mixed glue, or any combination thereof, such as printing silver glue first, and then printing into Carbon glue, which is a conductive material suitable for screen printing, is then dried and cured at 60-70 degrees Celsius.

In the (2) step of the method of the present invention, an electrically insulating flat plate with a thickness of more than 0.2 mm is covered on the same side surface of the conductive film with a double-sided adhesive pad insulating flat plate, and the partially exposed conductive film is reserved. To form an anode joint 307, a cathode joint 308, a reference electrode 309, and a working electrode 310, as shown in FIG. 7 . The groove channel area formed by raising the insulating plate 311 is the area 303 of the reaction film layer.

In step (3), the reaction reagent is coated on the surface of the reaction area in one procedure, as shown in FIG. 8 .

In step (4), cover the upper cover protection plate with a hole on the insulating plate 311 and form three openings in the reaction area, as shown in FIG. 9 .

The detection method of the electrode test piece of the present invention is quite easy. It is carried out in the electrochemical sensor of the sheet electrode. The reduction of active substances and electronic mediators produces a redox reaction, and the electrical effect generated by the reaction can be easily measured by using an electrochemical sensor to infer the concentration of the analyte.

The present invention also provides an electrochemical sensing device, which includes a disposable electrode test piece and a sensor, which can directly analyze the content of the analyte in the specimen.

Application example

On a flat surface of a polyethylene terephthalate (PET) insulating substrate 301, two mutually independent carbon glue conductive film anodes and cathodes are printed by screen printing. Then dry it at 60-80 degrees Celsius. Immediately, the same side surface of the existing conductive film is covered with a second layer of insulating flat plate 311 to form a reaction zone 303 of a groove channel, and a part of the exposed conductive film is reserved to form an anode joint 307, a cathode joint 308, Working electrode 310 and reference electrode 309 . The area formed by the groove channel is the area of the reaction film layer.

Afterwards, the reaction reagents containing the following formula components and ratios were added dropwise into the groove channel, and dried at 45 degrees Celsius. Then cover the upper protective plate 306 with a hole, the hole is located on the channel, and the electrochemical electrode test piece is completed.

Glucose oxidase (270U/mg) 0.7%

Carboxymethyl cellulose 3%

Potassium ferricvanide red blood salt (potassium ferricyanide) 7%

TritonX—100 0.08%

Phosphate buffer solution pH5.5; 0.1M 89.22%

The prepared electrochemical electrode test piece was measured for the whole blood sample, and it was found that the blood glucose concentration obtained by the glucose detection electrode test piece of the present invention was completely consistent with the result measured by the biochemical analyzer Ys12300. Refer to Figure 10.

Application Example 2

The step of application embodiment 1 is repeated, and the formula composition and the ratio of its reaction reagent are changed into following replacement:

Glucose oxidase (270U/mg) 0.6%

Albumin 1%

Methylcellulose 2%

Red blood salt (potassium ferricyanide) 6%

TritonX 100 0.09%

Phosphate buffer solution pH—5.5; 0.1M 90.31%

Application Example 3

The step of application embodiment 1 is repeated, and the formula composition and the ratio of its application reagent are changed into the following substitutions:

Uric acid oxidase (1000U/g) 1.5%

Methylcellulose 2.5%

Red blood salt 6.5%

Triton X—100 0.1%

Phosphate buffer solution pH8.5; 0.2M 89.41%

Application Example 4

The step of application embodiment 1 is repeated, and the formula composition and the ratio of its reaction reagent are changed into following replacement:

Glucose oxidase (270U/mg) 0.8%

Gelatin 2%

Albumin 0.5%

Red blood salt (potassium ferricyanide) 7.5%

TritonX—100 0.09%

Phosphate buffer solution pH: 5.5; 0.1M 89.11%

Application Example 5

The step of application embodiment 1 is repeated, and the formula composition and the ratio of its biological reaction agent are changed into the following substitutions:

Glucose oxidase (270U/mg) 0.66%

Albumin 0.5%

Polyvinyl azacyclone 2%

Red blood salt (potassium ferricyanide) 7%

Triton X—100 0.12%

Phosphate buffer solution pH: 5.5; 0.1M 89.53%.

Claims (40)

1. An electrochemical electrode test strip, comprising an electrically insulating flat substrate, a conductive film, a reaction film layer and a reaction zone space, characterized in that: the conductive film is positioned on the surface of the electrically insulating flat substrate to form an electrode system, The electrode system includes at least a working electrode and a reference electrode; the reaction film layer is formed on the surface of the reaction zone on the electrode system, and the reaction film contains at least one active material with redox properties, which is used to contact the detection The body produces a specific biochemical or chemical redox reaction; the reaction area is spaced on the electrically insulating flat substrate, and the surface on the same side as the conductive film is covered with a second layer of insulating flat to form a groove channel The reaction area is covered with an electrically insulating upper cover with holes to protect the plate, forming a space for carrying the reaction film and the sample, and has three openings. 2. The electrochemical electrode test strip according to claim 1, characterized in that: the electrical insulation substrate is selected from polyethylene terephthalate polyester board, glass board, polycarbonate board, polyvinyl chloride board, glass Any of fiberboard, bakelite board, ceramic board, polyester board. 3. The electrochemical electrode test strip according to claim 1, wherein the electrode system at least includes a working electrode and a reference electrode, which is a two-pole electrode system. 4. The electrochemical electrode test strip according to claim 1, wherein the electrode system comprises at least a working electrode, a reference electrode and an auxiliary electrode, and is a three-pole electrode system. 5. The electrochemical electrode test strip as claimed in claim 1, wherein the thickness of the second raised electrically insulating plate for forming the groove channel of fixed width is at least 0.2 mm. 6. The electrochemical electrode test strip according to claim 1, wherein the area of the reference electrode in the electrode system is greater than or equal to the area of the working electrode. 7. The electrochemical electrode test piece as claimed in claim 1, wherein the reaction film layer is used to contact with the sample to generate an electrochemical specific biochemical or chemical redox reaction. 8. The electrochemical electrode test strip according to claim 1, characterized in that: the size of the hole on the protective plate of the electrical insulation cover is equal to or larger than the width of the channel of the groove. 9. The electrochemical electrode test strip according to claim 1, characterized in that: two ends of the reagent reaction area of the electrode test strip protrude. 10. The electrochemical electrode test strip according to claim 1, characterized in that: the polymer contained in the reaction film component is selected from chitin, albumin, polyvinyl azapentacyclone, polyethylene Ethylene glycol, gelatin, dextran, polyvinyl alcohol, polyvinyl alcohol esters, methylcellulose, carboxymethylcellulose. 11. The electrochemical electrode strip as claimed in claim 1, wherein the reaction film component contains an electron mediator. 12. The electrochemical electrode test piece as claimed in claim 1, wherein the reaction film component contains a non-ionic surfactant. 13. The electrochemical electrode test strip according to claim 12, characterized in that: the non-ionic surfactant includes trinitrotoluene, lecithin, phosphatidylcholine, oleic acid, cyclodextrin, poly At least one of oxyethylene, glycerin, and fatty acid. 14. An electrochemical electrode test piece, which includes an electrically insulating flat substrate, a conductive film, a reaction film layer, a reaction zone space, and an upper cover protection plate, characterized in that the conductive film is located on the surface of the flat electrically insulating substrate Above, an electrode system, at least one working electrode, and a reference electrode are formed; the reaction film layer contains at least one active substance with redox properties, wherein the active substance refers to immobilized or unimmobilized redox enzymes , redox protein or redox content, which is used to produce a specific biochemical or chemical redox reaction with the contacted sample; the reaction area is covered with the second surface on the same side of the conductive film on the electrically insulating flat substrate. A layer of insulating flat plate is used to form the reaction area of the groove channel, and an electric insulating flat plate with holes and nitrocellulose membrane is covered to cover the protective flat plate to form a space for carrying the reaction film and the sample, and has three openings; The upper cover protection plate corresponds to the same side of the reaction area, and has the same size and holes as the channel of the groove with the nitrocellulose membrane. 15. The electrochemical electrode test strip according to claim 14, characterized in that the width of the nitrocellulose membrane is the same as that of the concave channel, and the thickness is at least 0.15 mm. 16. The electrochemical electrode test piece as claimed in claim 14, characterized in that: the electrically insulating upper cover protection plate has a nitrocellulose membrane of the same size as the channel of the groove, which is used to speed up the specimen covering the reaction area and reduce the time difference The resulting measurement error. 17. An electrochemical test piece, comprising an electrically insulating flat substrate layer, a conductive layer, a raised insulating layer, a reaction film layer, and an upper cover protection flat plate, characterized in that the conductive layer is formed on the flat insulating substrate On the surface, the conductive layer at least includes a working electrode and a reference electrode, and the working electrode and the reference electrode are not in contact with each other; the raised insulating layer covers the conductive film layer, wherein the raised insulating layer edge is recessed The groove channel is divided into a first spacer wall and a second spacer wall, and part of the conductive layer is exposed at the bottom of the groove channel; the reactive film layer covers the exposed conductive film layer of the groove channel; the upper cover protection plate covers On the raised insulation layer, the upper cover protection plate has a hole at the position corresponding to the channel; Specimen carrying space. 18. The electrochemical electrode test piece as claimed in claim 17, characterized in that: the components of the reaction film layer include active substances with redox properties, electronic mediators, high molecular polymers and non-ionic surfactants . 19. The electrochemical electrode test strip according to claim 18, characterized in that: the redox active substance is used to produce a specific biochemical or chemical redox reaction with the contacted sample, and is selected from the following components or Its combination: immobilized or unimmobilized redox enzymes, redox proteins or components with specific recognition ability in redox compounds. 20. The electrochemical electrode test strip according to claim 18, characterized in that: the polymer is selected from the following components or combinations thereof: chitin, albumin, polyvinyl azapentacyclone, polyethylene Glycol esters, gelatin, dextran, polyvinyl alcohol, polyethyl acetate, methylcellulose, carboxymethylcellulose. 21. The electrochemical electrode strip according to claim 18, wherein the electron mediator is selected from potassium ferricyanide. 22. The electrochemical electrode test strip according to claim 18, characterized in that: the non-ionic surfactant is selected from the following components: trinitrotoluene, lecithin, phosphatidylcholine, oleic acid, cyclodextrin , polyoxyethylene, glycerin, at least one of lipoic acid. 23. The electrochemical electrode test strip according to claim 17, wherein the conductive film comprises an auxiliary electrode. 24. The electrochemical electrode test strip according to claim 17, wherein the thickness of the insulating layer is at least 0.2 mm. 25. The electrochemical electrode test strip according to claim 17, wherein the area of the reference electrode is greater than or equal to the area of the working electrode. 26. The electrochemical electrode test strip according to claim 17, wherein the diameter of the hole is equal to or larger than the width of the channel of the groove. 27. The electrochemical electrode test strip according to claim 17, characterized in that: a layer of nitrocellulose film is provided on the side of the upper cover protection plate facing the reaction space area. 28. The electrochemical electrode test piece as claimed in claim 17, characterized in that: the upper cover protection plate contains a layer of nitrocellulose membrane. 29. The electrochemical electrode test strip according to claim 28, characterized in that the nitrocellulose membrane has the same width as the channel of the groove, and the thickness is at least 0.15 mm. 30. The electrochemical electrode test strip according to claim 17, characterized in that: the sample carrying space uses the hole of the upper cover protection plate as the first opening, and the channel in the groove is not covered by the first and second openings. The two ends blocked by the second partition wall are respectively the second opening and the third opening. 31. The electrochemical electrode test strip according to claim 30, wherein when the first opening is used for sample introduction, the second opening and the third opening are exhaust ports. 32. The electrochemical electrode test strip according to claim 30, wherein when the second opening is used for sample introduction, the first opening and the third opening are exhaust ports. 33. The electrochemical electrode test strip according to claim 17, characterized in that: the electrically insulating flat layer is selected from polyethylene terephthalate polyester board, polycarbonate board, polyvinyl chloride board, glass fiber board, Any of glass plate, bakelite plate, ceramic plate, polyester plate. 34. The electrochemical electrode test strip according to claim 17, characterized in that the width of the nitrocellulose membrane is the same as that of the channel of the groove, and the thickness is at least 0.15mm. 35. A method for manufacturing an electrochemical electrode test piece, characterized in that it comprises the following steps: First, an electrically insulating flat substrate is provided; a conductive film is covered on the upper surface of the electrically insulating flat substrate, so that the conductive film forms two mutually disconnected anode parts and cathode parts; a pad is covered on the conductive film An insulating layer, an opening is formed in the raised insulating layer, and the opening forms a groove channel, so that one end of the bare part of the conductive film anode part not covered by the raised insulating layer forms at least one reference electrode and the other end forms An anode joint, one end of the bare part of the cathode part of the conductive film that is not covered by the electrically insulating flat plate forms at least one working electrode, and forms a cathode joint with the other end; a reaction reagent is applied to the concave channel; drying is performed to make the reaction reagent A reaction film is formed to cover the bottom of the groove to form a reaction area; and an upper cover protection plate with a hole is covered on the raised insulating plate, and three openings are formed in the reaction area. 36. The electrochemical electrode test strip according to claim 35, characterized in that: the reaction reagent is composed of active substances with redox properties and a mixed solution, and the mixed solution includes electronic mediators, high molecular polymers, non- Composed of ionic surfactant and buffer solution. 37. The electrochemical electrode test piece as claimed in claim 36, characterized in that the weight percentage of the composition formula of the polymer reaction reagent is 1-4%. 38. The electrochemical electrode test piece as claimed in claim 36, characterized in that the electron mediator accounts for 1-10% by weight of the reaction reagent composition formula. 39. The electrochemical electrode test piece according to claim 36, characterized in that the non-ionic surfactant accounts for less than 0.3% by weight of the reaction reagent composition formula. 40. The electrochemical electrode test strip according to claim 36, characterized in that the buffer solution accounts for 83.7-98% by weight of the reaction reagent composition formula.

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