CN113588747A - Electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase - Google Patents
Electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase Download PDFInfo
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- 102100036475 Alanine aminotransferase 1 Human genes 0.000 title claims abstract description 16
- 108010082126 Alanine transaminase Proteins 0.000 title claims abstract description 16
- 108090000340 Transaminases Proteins 0.000 title claims abstract description 16
- 102000014898 transaminase activity proteins Human genes 0.000 title claims abstract description 16
- 238000006243 chemical reaction Methods 0.000 claims abstract description 155
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- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 1
- 239000004372 Polyvinyl alcohol Substances 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
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- 239000002390 adhesive tape Substances 0.000 description 1
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- GBDZMMXUOBAJMN-UHFFFAOYSA-K azane;ruthenium(3+);trichloride Chemical compound N.N.N.N.N.N.[Cl-].[Cl-].[Cl-].[Ru+3] GBDZMMXUOBAJMN-UHFFFAOYSA-K 0.000 description 1
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- 238000006467 substitution reaction Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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Abstract
The invention discloses an electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase, which comprises a PET (polyethylene terephthalate) substrate, a washout white glue layer, a hydrophilic film layer and a cover sheet layer which are sequentially arranged, wherein an electrode layer is attached to the PET substrate, a reaction area on the electrode layer is provided with a pair of reaction electrodes and a full blood electrode, the pair of reaction electrodes are positioned at one end close to a sampling port of the reaction area, the full blood electrode is positioned at one end far away from the sampling port of the reaction area, one end of the PET substrate far away from the reaction area is provided with pins, and the pins are correspondingly connected with the reaction electrodes and the full blood electrode one by one. The electrochemical test card has the advantages of small blood sampling amount, high detection speed and capability of eliminating the interference of endogenous pyruvic acid in fingertip blood on the detection of ALT and AST, so that the measurement result is more accurate.
Description
Technical Field
The invention belongs to the technical field of electrochemical test paper, and particularly relates to an electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase.
Background
Glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase are two important enzymes of human metabolism, and the measurement of ALT and AST in blood mainly reflects the sensitive indexes of liver injury, and various acute liver injuries can cause the elevation of ALT and AST. Therefore, the detection of ALT and AST concentrations in the human body allows targeted treatment according to the degree of impairment of the liver function of the human body.
At present, two kinds of transaminase in vivo need to be measured by a large biochemical analyzer and a professional in a hospital, or a test card for testing ALT and AST by single electrochemistry is adopted, the former increases the detection time and cost and is inconvenient to operate, and the latter cannot simultaneously measure ALT and AST, thus bringing inconvenience to measurement. Therefore, it is imperative to invent a method for simultaneously measuring ALT and AST.
Disclosure of Invention
Based on the technical defects, the invention provides the electrochemical test card for simultaneously measuring ALT and AST, which has the advantages of small blood collection amount, high detection speed and capability of eliminating the interference of endogenous pyruvic acid in fingertip blood on the detection of ALT and AST, so that the measurement result is more accurate.
In order to achieve the technical purpose, the invention specifically adopts the following technical scheme:
an electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase comprises a PET substrate, a washgroove white glue layer, a hydrophilic film layer and a cover plate layer which are sequentially arranged, wherein an electrode layer is attached to the PET substrate, a reaction area on the electrode layer comprises a first reaction area, a second reaction area and a third reaction area, a pair of reaction electrodes and a full blood electrode are respectively arranged on the first reaction area, the second reaction area and the third reaction area, the pair of reaction electrodes are positioned at one end close to a sampling port of the reaction area, the full blood electrode is positioned at one end far away from the sampling port of the reaction area, the pair of reaction electrodes of the first reaction area are used for detecting ALT and endogenous pyruvic acid, the pair of reaction electrodes of the second reaction area are used for detecting AST and endogenous pyruvic acid, the pair of reaction electrodes of the third reaction area are used for detecting full blood sample in the reaction area, pins are arranged at one end of the PET substrate far away from the reaction area, and the pins are connected with the reaction electrodes in a one-to-one correspondence manner.
The electrode layer is also provided with a starting electrode, the starting electrode is formed by direct short circuit of two electrodes, when the detection instrument is inserted, the two electrodes are disconnected with the detection instrument to conduct a circuit, and the instrument is automatically started.
The electrode layer is formed by printing conductive silver paste, conductive carbon paste and insulating ink on the PET sheet in a screen printing mode.
The first reaction zone, the second reaction zone and the third reaction zone are isolated by a notching white glue layer and form isolated sampling ports; first reaction zone, second reaction zone and third reaction zone coat have the enzyme, it has hydrophilic rete to coat on the white glue layer of dashing groove, is provided with the exhaust hole on hydrophilic rete, exhaust hole, enzyme and hydrophilic rete constitute siphon pond structure, and the blood sample is inhaled in the three independent reaction zone through the siphon effect from the sample connection during the blood sampling.
The hydrophilic film layer is covered with a cover sheet, and the cover sheet leaks out of the exhaust hole.
The reaction liquid stored in the first reaction zone comprises the following components in percentage by mass: 87% of buffer solution, 2% of alpha-ketoglutaric acid, 2% of L-alanine, 2% of sucrose, x-1001% of triton, 1% of cellulose, 3% of ruthenium hexammoniate chloride and 2% of pyruvate oxidase.
The reaction liquid stored in the second reaction zone comprises the following components in percentage by mass: 84% of buffer solution, 2% of L-aspartic acid, 2% of L-alanine, 2% of sucrose, 2% of triton x-1001%, 1% of cellulose, 3% of ruthenium hexammoniate chloride, 2% of oxaloacetate decarboxylase and 2% of pyruvate oxidase.
The reaction liquid stored in the third reaction zone comprises the following components in percentage by mass: 91% of buffer liquid medicine, 2% of sucrose, 2% of triton x-1001%, 1% of cellulose, 3% of ruthenium hexammoniate chloride and 2% of pyruvate oxidase.
The invention has the beneficial effects that:
1) according to the invention, the concentrations of ALT and AST are measured simultaneously through three independent reaction areas, and the interference of endogenous pyruvic acid is eliminated, so that the test is more accurate, the blood sampling amount is small, and the test speed is high;
2) the full blood electrode is arranged, when the blood sucking reaction area is full of the reaction area, the full blood electrode is conducted to start detection, if the blood is not full of the reaction area, the full blood electrode is not conducted, the blood is prompted to be not full of the reaction area, and the risk of misleading the blood by an inaccurate test result under the condition that the blood is not full of the reaction area and is not perceived in the test process can be completely avoided;
3) the silver electrode or the carbon electrode used by the invention is used for manufacturing the test card, and the used reagent is cheap, thereby greatly reducing the production cost.
Drawings
FIG. 1 is a schematic view of a layered structure of an electrochemical test card according to the present invention;
FIG. 2 is a schematic diagram of an electrode distribution for an electrode layer of an electrochemical test card according to the present invention;
FIG. 3 is a schematic diagram of the electrochemical test card according to the present invention;
FIG. 4 is a standard curve for ALT detection using the electrochemical test card of the present invention;
FIG. 5 is a standard curve for AST detection using the electrochemical test card of the present invention;
the PET substrate 1, the PET substrate 2, the notching white glue layer 3, the hydrophilic film layer 4 and the cover sheet layer; 5. a sampling port;
6. a first reaction area, 6-1, a first full blood electrode, 6-2, a first reaction electrode a, 6-3 and a first reaction electrode b;
7. a second reaction area 7-1, a second full blood electrode 7-2, a second reaction electrode a, 7-3 and a second reaction electrode b;
8. a third reaction area 8-1, a third full blood electrode 8-2, a third reaction electrode a, 8-3 and a third reaction electrode b;
9. and starting the electrode.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to specific embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides an electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase, which comprises a PET substrate 1, a notching white glue layer 2, a hydrophilic film layer 3 and a cover plate 4 from bottom to top in sequence as shown in figure 1.
As shown in figure 2, the PET substrate 1 of the invention is provided with an electrode layer, and the electrode layer consists of a first reaction electrode a 6-2, a first reaction electrode b 6-3, a second reaction electrode a 7-2, a second reaction electrode b 7-3, a third reaction electrode a 8-2, a third reaction electrode b 8-3, a first full blood electrode 6-1, a second full blood electrode 7-1, a third full blood electrode 8-1 and a starting electrode 9.
Wherein, the first reaction electrode a 6-2, the first reaction electrode b 6-3 and the first full blood electrode 6-1 are sequentially arranged on the first reaction area 6 from top to bottom, the second reaction electrode b 7-3, the second reaction electrode a 7-2 and the second full blood electrode 7-1 are arranged on the second reaction area 7 from top to bottom, the third reaction electrode a 8-2, the third reaction electrode b 8-3 and the third full blood electrode 8-1 are arranged on the third reaction area 8 from top to bottom, the first full blood electrode 6-1, the second full blood electrode 7-1 and the third full blood electrode 8-1 are respectively arranged at the bottoms of the first reaction area 6, the second reaction area 7 and the third reaction area 8 for detecting the full degree of the blood sample, when the blood sample is not fully absorbed, the full blood electrodes are not conducted, it is indicated that the blood is not full, and the defect of misleading the blood by an inaccurate test result under the condition of not full and being undetected in the test process can be completely avoided.
The first reaction electrode a 6-2 and the first reaction electrode b 6-3, the second reaction electrode b 7-3 and the second reaction electrode a 7-2, and the third reaction electrode a 8-2 and the third reaction electrode b 8-3 are respectively matched for use, and are respectively used for detecting ALT and endogenous pyruvic acid, AST and endogenous pyruvic acid, and endogenous pyruvic acid.
Pins corresponding to a first reaction electrode a 6-2, a first reaction electrode b 6-3, a second reaction electrode a 7-2, a second reaction electrode b 7-3, a third reaction electrode a 8-2, a third reaction electrode b 8-3, a first full blood electrode 6-1, a second full blood electrode 7-1 and a third full blood electrode 8-1 are arranged at the bottom of the PET substrate 1, are connected in a one-to-one correspondence manner and are used for being connected and conducted with a detection instrument.
The electrode layer is formed by adopting a screen printing mode through conductive silver paste, conductive carbon paste and insulating ink. The conductive carbon paste is used for preparing each reaction electrode, each full blood electrode and each pin, the conductive silver paste is used for preparing a lead for connecting the reaction electrodes, the full blood electrodes and the pins, and the insulating ink is attached to the circuit and leaks out of the reaction area and the pins.
The starting electrode 9 of the invention is arranged at the bottom of the PET substrate 1 and is formed by direct short circuit of two electrodes, when a detection instrument is inserted, the two electrodes are conducted with a power-off interruption circuit of the detection instrument, and the instrument is automatically started.
The electrodes are covered with insulating ink for isolating the interference among the electrodes, and the first reaction area 6, the second reaction area 7 and the third reaction area 8 at the end parts are leaked out, the tail parts of the reaction electrodes and the full blood electrodes on the PET substrate 1, and the starting electrode 9 for being conducted with each detection circuit of the detection instrument.
The first reaction zone 6, the second reaction zone 7 and the third reaction zone 8 are respectively covered with enzyme, a notching white glue layer 2 is arranged on the enzyme, and the first reaction zone 6, the second reaction zone 7 and the third reaction zone 8 are respectively and independently isolated by the notching white glue to form three sampling ports 5, so that the reaction zones are mutually independent and mutually conducted interference is eliminated.
The notching white glue layer 2 is respectively and sequentially provided with a hydrophilic film layer 3 and a cover plate 4. The hydrophilic film layer 3 and the cover plate 4 are respectively provided with exhaust holes corresponding to the first reaction area 6, the second reaction area 7 and the third reaction area 8, the exhaust holes are as close as possible to one side of each reaction area, which is as close as possible to the electrode, and the exhaust holes, the reaction area enzyme and the hydrophilic film layer 3 form a siphon pool.
The reaction liquid stored in the first reaction zone 6 of the present invention has the following components: 87% of buffer solution, 2% of alpha-ketoglutaric acid, 2% of L-alanine, 2% of sucrose, x-1001% of triton, 1% of cellulose, 3% of ruthenium hexammoniate chloride and 2% of pyruvate oxidase. The reaction zone is used for detecting ALT + endogenous pyruvic acid, L-alanine and alpha-ketoglutaric acid generate pyruvic acid under the action of ALT, and the pyruvic acid and an electron mediator ruthenium hexaammine chloride undergo redox reaction under the action of pyruvate oxidase to generate current.
The reaction liquid stored in the second reaction zone 7 of the present invention has the following components: 84% of buffer solution, 2% of L-aspartic acid, 2% of L-alanine, 2% of sucrose, 2% of triton x-1001%, 1% of cellulose, 3% of ruthenium hexammoniate chloride, 2% of oxaloacetate decarboxylase and 2% of pyruvate oxidase. The reaction area is used for detecting AST + endogenous pyruvic acid, L-aspartic acid and L-alanine generate pyruvic acid under the action of the AST, and the pyruvic acid reacts with an electron mediator ruthenium hexaammine chloride under the action of pyruvate oxidase to generate current.
The reaction liquid stored in the third reaction zone 8 of the present invention comprises the following components: 91% of buffer liquid medicine, 2% of sucrose, 2% of triton x-1001%, 1% of cellulose, 3% of ruthenium hexammoniate chloride and 2% of pyruvate oxidase. The reaction area is used for detecting endogenous pyruvic acid, and the endogenous pyruvic acid reacts with the electron mediator ruthenium hexaammine chloride under the action of pyruvate oxidase to generate current.
And respectively differentiating the currents generated by the ALT + endogenous pyruvic acid and the AST + endogenous pyruvic acid with the current generated by detecting the endogenous pyruvic acid, so as to eliminate the interference of the endogenous pyruvic acid, obtain the current generated by the reaction of the ALT and the AST, and obtain the concentrations of the ALT and the AST by using a standard curve of the current and the concentration.
As shown in FIG. 3, the electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase of the present invention has the advantages that when the reaction area is filled with blood, each full blood electrode and the reaction electrode are conducted, indicating that the blood absorption amount is sufficient; when the reaction area is not filled with blood, the reaction electrode generates current to conduct, the full blood electrode is not triggered, or the reaction electrode is not triggered, the full blood electrode is conducted, the blood suction amount of the reaction area is insufficient, and the detection instrument prompts that the blood suction amount is insufficient. When the blood sucking amount of the three reaction areas is enough, the currents generated by respective reactions are detected, the current values generated by pyruvic acid in the middle reaction area are subtracted from the currents generated in the ALT reaction area and the AST reaction area respectively, so that the interference of endogenous pyruvic acid can be eliminated, and the ALT concentration and the AST concentration are respectively obtained according to the standard curve relationship of the fitted current values and the concentrations.
In another embodiment of the present invention, the electrochemical test card of the present invention is prepared by the steps of:
1) taking a PET substrate, and printing conductive silver paste, conductive carbon paste and insulating ink on the PET substrate, wherein the electrodes comprise a reaction electrode, a full blood electrode and a starter electrode. The three groups of reaction electrodes and the full blood electrode are respectively arranged in three independent reaction areas to respectively test ALT + endogenous pyruvic acid content, AST + endogenous pyruvic acid content and endogenous pyruvic acid content, the starting electrode is formed by directly short-circuiting the two electrodes, and the instrument is automatically started when the instrument is inserted into a test card and detects that conduction current exists between the two electrodes. And covering the electrode with insulating ink to expose a reaction area and electrode pins with a certain area, thereby obtaining the substrate of the electrochemical test card for testing ALT and AST.
2) Preparing treating liquid containing polyvinyl alcohol, carrying out hydrophilic treatment on the reaction area by using the treating liquid, and drying in a drying tunnel. The drying temperature of the treatment liquid is 50 ℃, and the time is 4 min.
3) Three groups of reaction liquid medicines are respectively prepared, wherein the first group is the reaction liquid medicine for testing ALT + endogenous pyruvic acid, the second group is the reaction liquid medicine for testing AST + endogenous pyruvic acid, and the third group is the reaction liquid medicine for testing endogenous pyruvic acid. Respectively dropping the prepared three groups of reaction liquid medicines in corresponding reaction zones, and drying in a drying tunnel at 50 deg.C for 20-25 min.
The buffer solution of the three reaction solutions was 0.5mmol/L Tris buffer solution having a PH of 7.3.
In the first group of reaction liquid medicine, the buffer solution accounts for 87%, the alpha-ketoglutaric acid accounts for 2%, the L-alanine accounts for 2%, the cane sugar accounts for 2%, the surfactant triton x-100 accounts for 1%, the cellulose accounts for 1%, the electron mediator ruthenium hexamine chloride accounts for 3%, and the pyruvate oxidase accounts for 2%.
In the second group of reaction liquid medicine, the buffer solution accounts for 84 percent, the L-aspartic acid accounts for 2 percent, the L-alanine accounts for 2 percent, the sucrose accounts for 2 percent, the surfactant triton x-100 accounts for 1 percent, the cellulose accounts for 1 percent, the electron mediator ruthenium hexammoniachloride accounts for 3 percent, the oxaloacetate decarboxylase accounts for 2 percent, and the pyruvate oxidase accounts for 2 percent.
In the third group of reaction liquid medicine, the buffer liquid medicine accounts for 91 percent, the cane sugar accounts for 2 percent, the surfactant triton x-100 accounts for 1 percent, the cellulose accounts for 1 percent, the electron mediator ruthenium hexamine chloride accounts for 3 percent, and the pyruvate oxidase accounts for 2 percent.
4) And after drying, respectively covering the notching white glue layer, the hydrophilic film layer and the cover sheet. The material of the notching white glue layer is Sowess WH234343 type double-sided adhesive tape, the hydrophilic membrane is subjected to single-sided hydrophilic treatment, the 3M 9901P type is used, and the material of the cover plate is synthetic paper.
Example 1
5 samples of fresh whole blood with a gradient of ALT and AST were prepared, each concentration being divided into 3, the first being tested with a biochemical analyzer for ALT and AST concentrations, the second being used to confirm the ALT and AST standard curves on the test card, and the third being used to confirm the standard curves and then to compare the ALT and AST concentrations with the biochemical concentrations using the test card.
In the standard curve confirmation process, 10 times of repeated tests are respectively carried out on each concentration sample, the CV of the tested current value is within 3%, and the average value of the tested current of each concentration is shown in the following table 1:
TABLE 1 ALT and AST test currents at different concentrations
| Glutamic-pyruvic transaminase concentration (g/L) | 20 | 50 | 100 | 300 | 500 |
| Glutamic-oxaloacetic transaminase concentration (g/L) | 20 | 50 | 100 | 300 | 500 |
| First reaction zone Current (nA) | 236 | 287 | 364 | 636 | 962 |
| Second reaction zone Current (nA) | 225 | 268 | 358 | 645 | 958 |
| Third reaction zone Current (nA) | 203 | 197 | 201 | 198 | 200 |
| ALT corresponding current (nA) | 33 | 90 | 163 | 438 | 762 |
| AST corresponding current (nA) | 22 | 71 | 157 | 447 | 758 |
The biochemical concentrations of ALT and AST and the corresponding current values were fitted linearly, respectively, to obtain standard curves for ALT and AST (fig. 4 and 5). From the standard curve, it can be seen that the biochemical concentrations of ALT and AST and the corresponding current values are linear, and can reach 0.9987 and 0.9997 respectively.
A third blood sample was taken and ALT and AST were tested on the third blood sample using a different electrochemical test card of the present invention, with the results shown in tables 2 and 3.
TABLE 2 ALT test results
TABLE 3 AST test results
As can be seen from tables 2 and 3, the deviation between the concentration of ALT and AST tested by the test card and the biochemical concentration is very small, and the interference of endogenous pyruvic acid is eliminated by utilizing the difference principle, so that the test card can realize that one drop of blood can simultaneously measure the concentration of ALT and AST, and the accuracy of the result is high.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (7)
1. An electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase comprises a PET substrate, a notching white glue layer, a hydrophilic film layer and a cover plate layer which are sequentially arranged, and is characterized in that an electrode layer is attached to the PET substrate, a reaction area on the electrode layer comprises a first reaction area, a second reaction area and a third reaction area, a pair of reaction electrodes and a full blood electrode are respectively arranged on the first reaction area, the second reaction area and the third reaction area, the pair of reaction electrodes are positioned at one end close to a sampling port of the reaction area, the full blood electrode is positioned at one end far away from the sampling port of the reaction area, the pair of reaction electrodes of the first reaction area is used for detecting ALT and endogenous pyruvic acid, the pair of reaction electrodes of the second reaction area is used for detecting AST and endogenous pyruvic acid, and the pair of reaction electrodes of the third reaction area is used for detecting endogenous pyruvic acid, the full blood electrode is used for detecting the full condition of a blood sample in the reaction area, one end of the PET substrate far away from the reaction area is provided with pins, and the pins are connected with the reaction electrodes in a one-to-one correspondence mode.
2. The electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase according to claim 1, wherein the electrode layer is provided with a start-up electrode, and the start-up electrode is formed by direct short circuit of two electrodes.
3. The electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase according to claim 2, wherein the electrode layer is formed by printing conductive silver paste, conductive carbon paste and insulating ink on the PET sheet by screen printing.
4. The electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase according to claim 1, wherein the first reaction zone, the second reaction zone and the third reaction zone are separated by a white glue layer and form separated sampling ports.
5. The electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase of claim 1, wherein the hydrophilic film layer is provided with an air vent, and the air vent, the enzyme and the hydrophilic film layer form a siphon structure.
6. The electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase according to claim 5, wherein the hydrophilic membrane layer is covered with a cover sheet, and the cover sheet leaks out of the vent hole.
7. The electrochemical test card for simultaneously measuring glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase according to claim 1, wherein the electrochemical test card comprises the following components in percentage by mass:
the reaction liquid stored in the first reaction zone comprises the following components: 87% of buffer solution, 2% of alpha-ketoglutaric acid, 2% of L-alanine, 2% of sucrose, x-1001% of triton, 1% of cellulose, 3% of ruthenium hexammoniate chloride and 2% of pyruvate oxidase;
the reaction liquid stored in the second reaction area comprises the following components: 84% of buffer solution, 2% of L-aspartic acid, 2% of L-alanine, 2% of sucrose, 2% of triton x-1001%, 1% of cellulose, 3% of ruthenium hexammoniate chloride, 2% of oxaloacetate decarboxylase and 2% of pyruvate oxidase;
the reaction liquid stored in the third reaction zone comprises the following components: 91% of buffer liquid medicine, 2% of sucrose, 2% of triton x-1001%, 1% of cellulose, 3% of ruthenium hexammoniate chloride and 2% of pyruvate oxidase.
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