Quantitative detection system and method based on enzyme molecule response hydrogel in body fluid

Abstract

The invention discloses a quantitative detection system and method based on enzyme molecule response hydrogel in body fluid. The quantitative detection system comprises a substance response unit, a measurement display unit and a gas passage, the substance response unit comprises a confinement pipeline, the confinement pipeline is filled with acceptor hydrogel to form a valve switch, and target enzyme molecules are utilized to cut the acceptor hydrogel to change the breakthrough pressure intensity of the valve switch; and the measurement display unit is used for monitoring the change of the air pressure and outputting digital information. According to the method disclosed by the invention, the stimulation response effect between the target molecule and the receptor hydrogel is combined, and the target molecule is utilized to destroy a corresponding functional group on a molecular chain of the receptor hydrogel, so that the network structure of the hydrogel is collapsed to change the property of the hydrogel, and the breakthrough pressure intensity of gas passing through a confinement pipeline is further changed; the result can be displayed in the measurement display unit in real time. The method has the functional characteristics of low cost, small quantity of samples required for detection, safety, no danger, simplicity and convenience.

Description

Technical Field

[0001] The invention belongs to the technical field of biological enzyme detection, and in particular relates to a quantitative detection system and method based on enzyme molecule response hydrogel in body fluid. Background Art

[0002] Hydrogels are three-dimensional networks composed of cross-linked polymer chains that can absorb and retain large amounts of water while maintaining their structural stability. The physical and chemical properties of this material can be optimized by adjusting its composition and structure to suit specific testing needs. In addition, the ease of operation of hydrogels allows them to be easily integrated into various testing devices, a feature that makes hydrogels very suitable for laboratory and clinical settings. Due to their unique properties and flexible applicability, hydrogels are increasingly used in scientific research, especially in the fields of biological testing and medical diagnosis.

[0003] Existing enzyme molecule response hydrogel detection methods are complex and cumbersome, and such methods usually require a large amount of sample, which further limits the convenience and practicality of their application, and cannot be detected in human body fluids. Therefore, how to better achieve safe, accurate, stable, low-cost, and quantitative detection in human body fluids has become a huge challenge. Summary of the invention

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art, provide a quantitative detection system and method based on enzyme molecule response hydrogel in body fluids, and solve the problems in the above-mentioned background technology.

[0005] One of the technical solutions adopted by the present invention to solve its technical problems is: providing a quantitative detection system based on enzyme molecule response hydrogel in body fluids, including a material response unit and a measurement display unit; the material response unit includes a confined channel, the confined channel is filled with receptor hydrogel to form a valve switch, and the target enzyme molecules are used to cut the receptor hydrogel to change the breakthrough pressure of the valve switch; the measurement display unit is used to monitor the change of air pressure and output digital information.

[0006] In a preferred embodiment of the present invention, a gas passage is further included, and the confined area pipeline is tightly connected to the measurement and display unit through the gas passage.

[0007] In a preferred embodiment of the present invention, the measurement and display unit includes a pressure sensor, a source measurement module and a display module. The pressure sensor is used to obtain the air pressure value in the gas passage in real time. The source measurement module is electrically connected to the pressure sensor for powering the pressure sensor and reading the data of the pressure sensor. The display module is used to output digital information of the air pressure value or the breakthrough pressure value.

[0008] In a preferred embodiment of the present invention, the pressure sensor, the source measurement module and the display module are respectively connected to the confined pipe through a gas passage.

[0009] In a preferred embodiment of the present invention, the confined pipe is made of glass, plastic or metal, and has a shape of cylindrical tube, square tube, conical tube or triangular tube, and the diameter of the tube (at the narrowest point) is 0.5-2 mm.

[0010] In a preferred embodiment of the present invention, the target enzyme molecule is an enzyme molecule dissolved in body fluids, including but not limited to one of trypsin, chymotrypsin, pepsin, glutamine aminotransferase, cholinesterase, glutamine aminotransferase or lipase, etc., and the target enzyme molecule can destroy the hydrogel structure, thereby changing the breakthrough pressure of the gas passing through the confined pipe in the gas passage. The body fluid includes but is not limited to one of saliva, sweat, urine, serum, etc.

[0011] In a preferred embodiment of the present invention, the receptor hydrogel contains peptide bonds, ester bonds or amide bonds, and the target enzyme molecules change the rheological properties or mechanical properties of the receptor hydrogel by cutting the corresponding groups of the receptor hydrogel. The rheological properties of the receptor hydrogel include but are not limited to one of storage modulus, loss modulus, complex viscosity, etc.

[0012] The second technical solution adopted by the present invention to solve the technical problem is: providing a quantitative detection method based on enzyme molecule response hydrogel in body fluid, using the above-mentioned quantitative detection system based on enzyme molecule response hydrogel in body fluid. Specifically comprising the following steps:

[0013] 1) Prepare standard enzyme solutions with gradient concentrations and draw a standard curve;

[0014] 2) Add the target enzyme molecule to be tested and place the mixture after it fully reacts with the receptor hydrogel in a confined pipe, connect the confined pipe to the gas passage, monitor the change of breakthrough pressure in the response process in real time through the measurement display unit, and display digital information.

[0015] In a preferred embodiment of the present invention, step 1) prepares standard enzyme solutions with gradient concentrations (for example, prepares four standard enzyme solutions with concentrations of 0 μg / mL, 0.05 μg / mL, 0.1 μg / mL, and 0.15 μg / mL, and measures the breakthrough pressure based on these four standard solutions, with the horizontal axis being the enzyme concentration and the vertical axis being the breakthrough pressure, and fits them into a curve, which is the standard curve).

[0016] In a preferred embodiment of the present invention, in step 2), the pressure change is monitored in real time by the pressure sensor of the measuring display unit, the breakthrough pressure is calculated, and the concentration of the target enzyme molecule is quantitatively calculated according to the concentration corresponding to the standard curve at this time.

[0017] This technical solution uses enzyme molecules to induce changes in the properties of receptor hydrogels, thereby causing changes in the breakthrough pressure of gases passing through macroscopic confined pipes to detect enzyme molecules. The confined pipe provides a simple and convenient space for hydrogel detection. The properties of the receptor hydrogel will change with the concentration of the target molecule, and its properties can be represented by the breakthrough pressure. Finally, the concentration of the target molecule can be judged based on the change in the breakthrough pressure. Compared with the background technology, it has the following advantages:

[0018] 1. The present invention is based on converting the qualitative and quantitative responses between the target molecule and the receptor hydrogel into breakthrough pressure changes when the gas passes through the macroscopic confined channel.

[0019] 2. The present invention provides a method for converting the reaction signal between biological macromolecules and gel molecular chains into a physical signal that can be directly displayed.

[0020] 3. The system provided by the present invention has the characteristics of low cost, portability, and small amount of sample required for detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is a schematic diagram of the structure of the quantitative detection system;

[0022] Among them: 1-measurement display unit, 2-gas passage, 3-material response unit, 1-1-display unit, 1-2-pressure sensor, 1-3-source measurement unit.

[0023] Figure 2 It is a schematic diagram of the principle of an embodiment;

[0024] Figure 3 Schematic diagram of the relationship between the breakthrough pressure of the enzyme molecule-containing hydrogel and the enzyme molecule concentration of Example 1;

[0025] Figure 4 This is a schematic diagram of the relationship between the breakthrough pressure of the hydrogel containing enzyme molecules and the concentration of enzyme molecules in urine in Example 2;

[0026] Figure 5 This is a schematic diagram of the relationship between the breakthrough pressure of the enzyme molecule-containing hydrogel and the enzyme molecule concentration in serum in Example 3. DETAILED DESCRIPTION

[0027] The present invention is further explained below in conjunction with the accompanying drawings and embodiments.

[0028] refer to Figure 1A quantitative detection system of hydrogels in body fluids based on enzyme molecule response comprises a measurement display unit 1, a gas passage 2, a material response unit 3, a display unit 1-1, a pressure sensor 1-2, and a source measurement unit 1-3. The display unit 1-1, the pressure sensor 1-2, and the source measurement unit 1-3 are integrated into a measurement display unit 1, and are all connected to the gas passage 2. The gas passage 2 and the material response unit 3 are connected, and the entire device has good airtightness. The target molecules and receptor hydrogels to be measured are placed in a macroscopic confined pipeline. When the gas passes through, the gas with sufficient pressure will destroy the structure of the hydrogel, and a breakthrough pressure P will appear. The process at this time will be recorded by the pressure sensor 1-2 and the display unit 1-1. The higher the concentration of the target molecules, the worse the storage modulus, complex viscosity, mechanical properties, etc. of the receptor hydrogel, the easier it is for the gas to destroy the structure of the hydrogel, and the smaller the breakthrough pressure P.

[0029] In the following embodiments, the breakthrough pressure of the target molecule of the same concentration in different body fluids will also be different. The receptor hydrogel is at least one of the groups containing peptide bonds, ester bonds, amide bonds, etc. The target (enzyme) molecule includes but is not limited to one of trypsin, chymotrypsin, pepsin, glutamine aminotransferase, cholinesterase, glutamine aminotransferase, lipase, etc. The target molecule can destroy the hydrogel structure, thereby changing the breakthrough pressure of the gas through the macroscopic confinement pipeline in the gas path. The body fluid includes but is not limited to one of saliva, sweat, urine, serum, etc.

[0030] The material of the macro-confined pipe includes but is not limited to one of a glass tube, a plastic tube, a metal tube, etc., the shape of the macro-confined pipe includes but is not limited to one of a cylindrical tube, a square tube, a conical tube, a triangular tube, etc., and the size of the macro-confined pipe (the narrowest point) includes one of 0.5 mm-2 mm.

[0031] The quantitative detection principle of the hydrogel based on enzyme molecule response in body fluids is as follows Figure 2 In the material response unit 3, when the target molecule to be tested is not added, the internal network structure of the receptor hydrogel is not destroyed, the mechanical strength and storage modulus of the hydrogel are relatively large, the gas cannot pass through the gas passage 2, and can only accumulate continuously. The measurement display unit 1 shows a high breakthrough pressure value. When the target molecule to be tested is added, the target molecule destroys the internal network structure of the receptor hydrogel, reducing its mechanical properties and storage modulus, and the gas will pass through the gas passage. At this time, there will be a breakthrough pressure value, and the concentration of the target molecule can be judged according to the breakthrough pressure.

[0032] Example 1

[0033] by Figure 1Based on the structure shown in the figure, gelatin is selected as the receptor hydrogel, trypsin is selected as the target molecule, and buffer is selected as the solvent of the trypsin solution. Trypsin of different concentrations is first dissolved in the buffer, and then added to a gelatin solution of a certain concentration and a certain volume. After fully reacting for a certain period of time, 10 microliters of the mixed solution is placed in a macroscopic confinement channel and allowed to stand for a period of time. The macroscopic confinement channel containing the mixed solution is placed in a gas passage, and the gas in the gas passage passes through the macroscopic confinement channel. The breakthrough pressure value obtained by the measurement display unit can be used to obtain the concentration of trypsin, as shown in FIG. Figure 3 .

[0034] Example 2

[0035] Gelatin is selected as the receptor hydrogel, trypsin is selected as the target molecule, and artificial urine is selected as the solvent of the trypsin solution. Trypsin of different concentrations is first dissolved in artificial urine, and then added to a gelatin solution of a certain concentration and a certain volume. After sufficient reaction for a certain period of time, 10 microliters of the mixed solution is placed in a macroscopic confinement channel, and allowed to stand for a period of time. The macroscopic confinement channel containing the mixed solution is placed in a gas passage, and the gas in the gas passage passes through the macroscopic confinement channel. The breakthrough pressure value obtained by the measurement display unit can be used to obtain the concentration of trypsin, such as Figure 4 .

[0036] Example 3

[0037] Gelatin is selected as the receptor hydrogel, trypsin is selected as the target molecule, and human serum is selected as the solvent of the trypsin solution. Trypsin of different concentrations is first dissolved in serum, and then added to a gelatin solution of a certain concentration and volume. After sufficient reaction for a certain period of time, 10 microliters of the mixed solution is placed in a macroscopic confinement channel, and allowed to stand for a period of time. The macroscopic confinement channel containing the mixed solution is placed in a gas passage, and the gas in the gas passage passes through the macroscopic confinement channel. The breakthrough pressure value obtained by the measurement display unit can be used to obtain the concentration of trypsin, such as Figure 5 .

[0038] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some or all of the technical features thereof may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A quantitative detection system based on enzyme molecule response hydrogel in body fluids, characterized by: It comprises a material response unit and a measurement and display unit; the material response unit comprises a confined channel, the confined channel is filled with a receptor hydrogel to form a valve switch, and the target enzyme molecules are used to cut the receptor hydrogel to change the breakthrough pressure of the valve switch; the measurement and display unit is used to monitor the change of air pressure and output digital information.

2. The quantitative detection system based on enzyme molecule responsive hydrogel in body fluid according to claim 1, characterized in that: It also includes a gas passage, and the confined area pipeline is tightly connected with the measurement and display unit through the gas passage.

3. The quantitative detection system based on enzyme molecule responsive hydrogel in body fluid according to claim 2, characterized in that: The measurement and display unit includes a pressure sensor, a source measurement module and a display module. The pressure sensor is used to obtain the air pressure value in the gas passage in real time. The source measurement module is electrically connected to the pressure sensor. The display module is used to output the measured data.

4. The quantitative detection system based on enzyme molecule responsive hydrogel in body fluid according to claim 3, characterized in that: The pressure sensor, source measurement module and display module are respectively connected to the confined area pipeline through a gas passage.

5. The quantitative detection system based on enzyme molecule responsive hydrogel in body fluid according to claim 1, characterized in that: The confined area pipe is made of glass, plastic or metal, and has a shape of cylindrical tube, square tube, conical tube or triangular tube, with a tube diameter of 0.5-2 mm.

6. The quantitative detection system based on enzyme molecule responsive hydrogel in body fluid according to claim 1, characterized in that: The target enzyme molecule is an enzyme molecule dissolved in body fluids, including trypsin, chymotrypsin, pepsin, glutamine aminotransferase, cholinesterase, glutamine aminotransferase or lipase, and the body fluids include saliva, sweat, urine, and serum.

7. The quantitative detection system based on enzyme molecule responsive hydrogel in body fluid according to claim 1, characterized in that: The receptor hydrogel contains peptide bonds, ester bonds or amide bond groups, and the target enzyme molecules change the rheological properties or mechanical properties of the receptor hydrogel by cutting the corresponding groups of the receptor hydrogel.

8. A quantitative detection method based on enzyme molecule response hydrogel in body fluids, characterized in that: A quantitative detection system based on enzyme molecule responsive hydrogel in body fluid as described in any one of claims 1 to 7 is used.

9. The quantitative detection method based on enzyme molecule responsive hydrogel in body fluid according to claim 8, characterized in that: The steps include: 1) Prepare standard enzyme solutions with gradient concentrations and draw a standard curve; 2) Add the target enzyme molecule to be tested and place the mixture with the receptor hydrogel in a confined pipe, connect the confined pipe to the gas passage, measure the breakthrough pressure during the response process by measuring the display unit, and calculate the concentration of the target enzyme molecule to be tested using the displayed digital information.

10. The quantitative detection method based on enzyme molecule responsive hydrogel in body fluid according to claim 9, characterized in that: In step 2), the pressure change is monitored in real time by the pressure sensor of the measuring display unit to obtain the breakthrough pressure, and the concentration of the target enzyme molecule is quantitatively calculated by comparing with the standard curve.

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