Glycosylated hemoglobin concentration detection device and method based on terahertz metamaterial

Abstract

The invention discloses a glycosylated hemoglobin concentration detection device and method based on a terahertz metamaterial. The glycosylated hemoglobin concentration detection device comprises a sample feeding and discharging module, a measuring light path module, a terahertz metamaterial detection module and a data processing and analyzing module, the terahertz metamaterial detection module comprises a stainless steel metal panel, a quartz transparent detection window, a Teflon sealing gasket, a polyethylene gasket with a prismatic notch groove in the center and a terahertz metamaterial chip. According to the constructed glycosylated hemoglobin concentration detection device, a detection sample is sucked into a terahertz metamaterial detection module through a peristaltic pump, glycosylated hemoglobin in the blood sample can be accurately captured by an antibody on the surface of a terahertz metamaterial chip, red shift of a terahertz transmission spectrum of the glycosylated hemoglobin is caused, and then a terahertz time-domain spectrometer is used for monitoring. Through the comparison curve of the detection concentration of the protein and the terahertz transmission spectrum, the detection of the concentration of the glycosylated hemoglobin in the blood of the diabetic patient can be conveniently and quickly realized.

Description

Technical Field

[0001] The present invention relates to the field of biological detection technology, and in particular to a device and method for detecting glycated hemoglobin concentration based on terahertz metamaterials. Background Art

[0002] Terahertz waves refer to electromagnetic waves with a wavelength range of 0.1 THz to 10 THz and a corresponding wavelength range of 0.03mm to 3mm. It has both photon-like and electron-like properties, so it has great application value and potential, especially in biomedicine. Metamaterials are artificial electromagnetic materials with properties that natural materials do not have. Terahertz metamaterials have shown great potential in biological detection, especially in the high-sensitivity sensing and identification of biological samples, providing advanced technical means for early diagnosis and treatment of major diseases.

[0003] Glycated hemoglobin is a combination of glucose and hemoglobin in human blood vessels. Short-term fluctuations in blood sugar levels cannot affect the production of this substance, and its stability in the blood is relatively high. The results of the glycated hemoglobin test can indicate the average blood sugar concentration of an individual two months before the test. In clinical practice, it is generally used as a key indicator to reflect the treatment of the disease during diabetes treatment.

[0004] Since different proteins have different dielectric constants, different proteins in the blood cause different frequency changes in metamaterials. By measuring the relationship curve between the concentration of a specific protein and the transmission red frequency shift or amplitude change in advance, a protein concentration comparison card can be obtained. When measuring a specific protein in the blood, you only need to correspond the transmission peak frequency shift or amplitude change to the previously obtained concentration comparison card to get the concentration of the protein.

[0005] At present, the clinical detection methods for the concentration of glycated hemoglobin in blood samples mainly use some biochemical methods, such as Western blot tracking (WB), enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay (IFA), etc. However, these methods cannot meet the current clinical needs for rapid detection. Therefore, it is necessary to develop a more sensitive, easier to operate and relatively shorter detection device. Summary of the invention

[0006] The purpose of the present invention is to provide a device and method for detecting the glycated hemoglobin concentration based on terahertz metamaterials, so as to solve the technical problem in the prior art that the glycated hemoglobin concentration in a blood sample cannot be detected quickly.

[0007] To achieve the above object, the present invention provides the following technical solutions: The terahertz metamaterial-based glycated hemoglobin concentration detection device provided by the present invention comprises a sample introduction and output module, a measurement optical path module, a terahertz metamaterial detection module, and a data processing and analysis module.

[0008] Furthermore, the sample introduction and sample output module includes a container and a peristaltic pump capable of controlling the flow rate of the liquid, and the container is used to hold the sample to be tested and the waste liquid.

[0009] Furthermore, the containers are two beakers, which are used to contain the sample to be tested and the waste liquid respectively, and the peristaltic pump is used to extract the sample and discharge the waste liquid into the beaker.

[0010] Furthermore, the measurement optical path module includes a terahertz time-domain spectrometer, a terahertz wave transmitting port, four terahertz wave-specific plano-convex lenses with a focal length of 50 mm, a terahertz wave receiving port, a 240 mm long dovetail guide rail, and an optical breadboard, and the optical path system is fixed on the optical breadboard through the guide rail.

[0011] Furthermore, the terahertz metamaterial detection module includes front and rear stainless steel metal panels, a pair of quartz transparent detection windows, a pair of Teflon sealing gaskets, a polyethylene gasket with a prismatic groove in the center, and a terahertz metamaterial chip. The terahertz metamaterial chip is located in the hollow area of ​​the polyethylene gasket, and is covered with transparent detection windows on the upper and lower parts, and the outer side of the transparent detection window is fixed by a central control metal frame bracket; the pair of Teflon sealing gaskets are respectively located between the transparent detection windows and between the lower detection window and the hollow metal frame bracket; the stainless steel metal front panel and the upper layer of the transparent detection window are provided with a through sample inlet and sample outlet.

[0012] Furthermore, the data processing and analysis module includes a host computer, and the host computer is connected to the terahertz time-domain spectrometer.

[0013] Furthermore, the data processing and analysis module includes a laptop computer, which is connected to the terahertz time-domain spectrometer via a data cable.

[0014] Furthermore, the terahertz metamaterial chip is made of a resonant ring array etched from a planar gold film on a quartz glass substrate.

[0015] Furthermore, the unit structure of the terahertz metamaterial array is composed of a pure gold square open resonant ring plus a pure gold square structure, the outer square side length of the square open ring is 20 μm, and the inner square side length is 15 μm; the length of the opening is 5 μm and the width is 5 μm.

[0016] Furthermore, the side length of the square pure gold structure in the unit structure of the terahertz metamaterial array is 5 μm.

[0017] Furthermore, a glycated hemoglobin-specific antibody is attached to the surface of the terahertz metamaterial chip to capture the glycated hemoglobin in the blood sample.

[0018] The present invention also provides a method for detecting the concentration of glycosylated hemoglobin based on terahertz metamaterials, comprising the following steps: (1) Prepare 9 samples of glycated hemoglobin with different concentration gradients. After loading, turn on the peristaltic pump switch and adjust the flow rate to 100 μL / min, and draw the sample into the terahertz metamaterial test module for antigen-antibody binding reaction for 20 minutes; open the sample outlet of the terahertz metamaterial test module, adjust the flow rate of the peristaltic pump to 10 μL / min, and pass PBS buffer to clean the inner walls of all pipes and the inside of the terahertz metamaterial test module for 5 minutes; finally, remove the PBS buffer from the sample inlet, and pump air into the peristaltic pump to remove the liquid on the chip surface in the terahertz metamaterial test module; turn on the terahertz spectrometer for testing; (2) Analyzing and fitting the measured data by a host computer connected to the terahertz time-domain spectrometer, obtaining a relationship curve between the glycated hemoglobin concentration and the shift of the resonance peak, and obtaining a glycated hemoglobin standard concentration comparison card; (3) The blood collected from the diabetic patient was added to the sample loading container. Before loading, the sample outlet of the terahertz metamaterial test module was closed. The peristaltic pump switch was turned on and the flow rate was adjusted to 100 μL / min. After that, the peristaltic pump sucked the sample into the terahertz metamaterial test module for antigen-antibody binding reaction for 20 minutes. The sample outlet of the terahertz metamaterial test module was opened, the flow rate of the peristaltic pump was adjusted to 10 μL / min, and PBS buffer was introduced to flush out the remaining components in the blood sample on the surface of the terahertz metamaterial chip, and the inner walls of all pipes and the inside of the terahertz metamaterial test module were cleaned for 5 minutes. Finally, the PBS buffer was removed from the sample inlet, and air was pumped into the terahertz metamaterial test module through the peristaltic pump to remove the liquid on the chip surface. The terahertz spectrometer was turned on for testing, and the amount of transmission peak shift was matched to the previous comparison card to obtain the glycated hemoglobin concentration in the patient's blood.

[0019] Based on the above technical solution, the embodiments of the present invention can at least produce the following technical effects: The present invention provides a glycated hemoglobin concentration detection device and method based on terahertz metamaterials. The chip of the terahertz metamaterial detection module is made of a resonant ring array etched by a planar gold film on a quartz glass substrate and has glycated hemoglobin-specific antibodies on the surface. The constructed glycated hemoglobin concentration detection device sucks the detection sample into the terahertz metamaterial detection module through a peristaltic pump. The antibodies on the surface of the terahertz metamaterial chip can accurately capture the glycated hemoglobin in the blood sample, causing its terahertz transmission spectrum to red shift, and then monitor it using a terahertz time-domain spectrometer. Through the comparison curve of the detection concentration of the protein and the terahertz transmission spectrum, the glycated hemoglobin concentration in the blood of diabetic patients can be detected conveniently and quickly. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the structures shown in these drawings without paying creative work.

[0021] Figure 1 This is a structural diagram of a terahertz metamaterial chip according to an embodiment of the present invention; Figure 2 This is a schematic diagram of the testing principle of a terahertz metamaterial chip according to an embodiment of the present invention; Figure 3 is a simulated transmission spectrum of the terahertz metamaterial chip according to an embodiment of the present invention; Figure 4 This is a schematic diagram of a device for detecting glycated hemoglobin using a terahertz metamaterial according to an embodiment of the present invention; Figure 5 This is an assembly diagram of a terahertz metamaterial detection module according to an embodiment of the present invention; In the figure: 1-glycosylated hemoglobin; 2-antibody; 3-metamaterial structure; 4-substrate; 5-all-fiber terahertz time-domain spectrometer; 6-beaker 1; 7-peristaltic pump; 8-optical platform; 9-laptop computer; 10-beaker 2; 11-x-axis dovetail guide slider; 12-y-axis dovetail guide slider; 13-optical breadboard; 14-dovetail guide; 15-terahertz wave transmitting end; 16-a plano-convex lens 1 with a focal length of 50 mm; 17-a plano-convex lens 2 with a focal length of 50 mm; 18- Liquid sample pool; 19- three plano-convex lenses with a focal length of 50 mm; 20- four plano-convex lenses with a focal length of 50 mm; 21- terahertz wave receiving end; 22- liquid filling port; 23- nut; 24- stainless steel front panel; 25- Teflon gasket one; 26- perforated quartz glass transparent window; 27- terahertz metamaterial chip; 28- polyethylene gasket; 29- quartz glass transparent window; 30- Teflon gasket two; 31- stainless steel rear panel; 32- threaded column; 33- stainless steel fixing plate. DETAILED DESCRIPTION

[0022] The following will combine the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work belong to the scope of protection of the present invention.

[0023] In the description of the present application, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inside", "outside" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present application; the terms "first", "second", and "third" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance; in addition, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be a connection between the two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present application can be understood according to specific circumstances.

[0024] Example 1: Preparation of terahertz metamaterial chip A titanium layer with a thickness of 10 nm and a gold layer with a thickness of 150 nm were deposited on a quartz glass substrate with a dielectric constant of 3.9 by magnetron sputtering. The DC conductivity of gold is 4.56×107 S / m. Then, projection step lithography is used to prepare the corresponding metal pattern, where the specific shape is as follows Figure 1 As shown. Each unit of the array is a square with a side length P of 40μm. The metal pattern is composed of a square open resonant ring plus a square structure. The side length L1 of the outer square of the square open ring is 20μm, and the side length L2 of the inner square is 15μm. The length g of the opening is 5μm, and the side length L of the square structure is 5μm. The terahertz metamaterial array is only sensitive to x-polarization. The specific parameters are specially designed for glycated hemoglobin, which can maximize the detection capability of terahertz metamaterials. The x-polarization transmission spectrum of terahertz metamaterials is shown in Figure 3 As shown; Terahertz metamaterial chips Figure 1 As shown; The terahertz metamaterial chip is composed of a substrate 4, a metamaterial structure 3, and an antibody 2, and the antibody 2 is bound to glycated hemoglobin; Figure 2 As shown, a specific antibody or probe of glycated hemoglobin is modified on the surface of the terahertz metamaterial, the concentration of the antibody or probe is controlled to be 5-10 μg / mL, and then it is placed in a terahertz metamaterial test module.

[0025] Example 2: Construction of a glycosylated hemoglobin concentration detection device based on terahertz metamaterials like Figure 4 Shown, including (1) Sample injection and sample output module: Beaker 1 6: a beaker with unlimited range and sterilized in advance at high temperature, used to hold the blood sample to be tested; Beaker 2 10: a beaker with unlimited range, used to hold waste liquid; a pipe with an outer diameter of 3 mm and an inner diameter of 1 mm is connected to the sample inlet of the terahertz metamaterial detection module, leading to the sample inlet beaker; a pipe with an outer diameter of 3 mm and an inner diameter of 1 mm is connected to the sample outlet of the terahertz metamaterial detection module, leading to the waste liquid tank; a peristaltic pump 7 with adjustable flow rate.

[0026] (2) Measurement optical path module: The all-fiber terahertz time-domain spectrometer 5 generates effective terahertz waves in the effective frequency range of 0-3 terahertz; the entire optical path system is located at the upper end of an optical breadboard 13, and all optical components are fixed on a dovetail guide 14 with a length of 240 mm through a dovetail guide slider that can move in the xy axis; from the terahertz transmitting end 15 to the terahertz receiving end 21, the other components in the middle are: a plano-convex lens 16 with a focal length of 50 mm, a convex section of the lens facing the receiving port side; a plano-convex lens 2 17 with a focal length of 50 mm, a convex section of the lens facing the transmitting port side; a terahertz metamaterial test module, with a side with a sample inlet and outlet port facing the transmitting port side; a plano-convex lens 3 19 with a focal length of 50 mm, a convex section of the lens facing the receiving port side; a plano-convex lens 4 20 with a focal length of 50 mm, a convex section of the lens facing the transmitting port side.

[0027] (3) Terahertz metamaterial detection module: The terahertz metamaterial detection module is as follows: Figure 5 As shown, it includes front and rear stainless steel metal panels, a pair of transparent detection windows made of quartz material, a pair of Teflon sealing gaskets, a polyethylene gasket with a prismatic groove in the center and a terahertz metamaterial chip; the stainless steel front panel 24, Teflon gasket 25, perforated quartz glass transparent window 26, terahertz metamaterial chip 27, polyethylene gasket 28, quartz glass transparent window 29, Teflon gasket 30, and stainless steel rear panel 31 are assembled in sequence and fixed by nuts 23 and threaded columns 32, and there is also a liquid filling port 22 above the stainless steel front panel 24.

[0028] The terahertz metamaterial chip is located in the hollow area of ​​the polyethylene gasket, and is covered with transparent detection windows on the upper and lower covers. The outer side of the transparent detection window is fixed by a central control metal frame bracket; the pair of Teflon sealing gaskets are respectively located between the transparent detection windows and between the lower detection window and the hollow metal frame bracket; the stainless steel metal front panel and the upper layer of the transparent detection window are provided with a through sample inlet and a sample outlet.

[0029] (4) Data processing and analysis module: The main component of this module is a laptop computer equipped with the terahertz time-domain spectrometer operating software, which is connected to the terahertz time-domain spectrometer via a data cable.

[0030] Example 3: Using a terahertz metamaterial-based glycosylated hemoglobin concentration detection device to detect glycosylated hemoglobin concentration, the specific steps are as follows: (1) Prepare 9 samples of glycated hemoglobin with different concentration gradients, namely 1 ng / mL, 5 ng / mL, 10 ng / mL, 50 ng / mL, 100 ng / mL, 200 ng / mL, 400 ng / mL, 800 ng / mL, and 1000 ng / mL, and their volume is 70 μL; then place them in different beakers; close the sample outlet of the terahertz metamaterial test module before loading; turn on the peristaltic pump switch and adjust the flow rate to 100 μL / min, then the peristaltic pump draws the samples into the terahertz metamaterial test module in turn for antigen-antibody binding reaction for 20 min; open the sample outlet of the terahertz metamaterial test module, adjust the flow rate of the peristaltic pump to 10 μL / min, and pass PBS buffer to clean the inner walls of all pipes and the inside of the terahertz metamaterial test module for 5 min; finally, remove the PBS buffer from the sample inlet, pump air into the peristaltic pump to remove the liquid on the chip surface in the terahertz metamaterial test module; turn on the terahertz spectrometer for testing.

[0031] (2) As the concentration of glycated hemoglobin increases, frequency shifts of varying degrees occur compared to the transmission spectra of the terahertz metamaterial whose surface is only modified with glycated hemoglobin-specific antibodies. The glycated hemoglobin concentration and the shift of the resonance peak show a specific functional correlation. The measured data are analyzed and fitted by computer to obtain a relationship curve between the two and a glycated hemoglobin standard concentration comparison card. (3) Add the blood collected from the diabetic patient to the beaker containing the sample, and close the sample outlet of the terahertz metamaterial test module before loading; turn on the peristaltic pump switch and adjust the flow rate to 100 μL / min, then the peristaltic pump draws the sample into the terahertz metamaterial test module for antigen-antibody binding reaction for 20 minutes; open the sample outlet of the terahertz metamaterial test module, adjust the flow rate of the peristaltic pump to 10 μL / min, pass PBS buffer to flush out the remaining components in the blood sample on the surface of the terahertz metamaterial chip, and clean all the inner walls of the pipes and the inside of the terahertz metamaterial test module for 5 minutes; finally, remove the PBS buffer from the sample inlet, and pump air into the terahertz metamaterial test module through the peristaltic pump to remove the liquid on the chip surface in the terahertz metamaterial test module; turn on the terahertz spectrometer for testing, and correspond the amount of transmission peak shift to the previous comparison card to obtain the glycated hemoglobin concentration in the patient's blood.

[0032] The structures, functions and connection forms disclosed in this article can be implemented in other ways. For example, the embodiments described above are only exemplary, for example, multiple components can be combined or integrated into another component; in addition, each functional component in each embodiment of this article can be integrated into one functional component, or each functional component can exist physically separately, or two or more functional components can be integrated into one functional component.

[0033] The above is only a preferred embodiment of the present invention. It should be pointed out that a person skilled in the art can make several modifications and improvements without departing from the inventive concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A glycosylated hemoglobin concentration detection device based on terahertz metamaterials, characterized in that: It includes a sample feeding and discharging module, a measurement optical path module, a terahertz metamaterial detection module, and a data processing and analysis module; the terahertz metamaterial detection module includes a stainless steel metal panel, a quartz transparent detection window, a Teflon sealing gasket, a polyethylene gasket with a prismatic groove in the center, and a terahertz metamaterial chip.

2. The terahertz metamaterial-based glycated hemoglobin concentration detection device according to claim 1, characterized in that: The sample injection and sample output module comprises a container and a peristaltic pump capable of controlling the flow rate of the liquid, and the container is used for containing samples to be tested and waste liquid.

3. The glycated hemoglobin concentration detection device based on terahertz metamaterial according to claim 1, characterized in that: The measuring optical path module consists of a terahertz time domain spectrometer, a terahertz wave transmitting port, four plano-convex lenses with a focal length of 50 mm, a terahertz wave receiving port, a 240 mm long dovetail guide rail, and an optical breadboard. The optical path system is fixed on the optical breadboard through the guide rail.

4. The glycated hemoglobin concentration detection device based on terahertz metamaterial according to claim 1, characterized in that: The terahertz metamaterial chip is located in the hollow area of ​​the polyethylene gasket, and is covered with transparent detection windows on the upper and lower parts. The outer side of the transparent detection window is fixed by a hollow metal frame bracket; the pair of Teflon sealing gaskets are respectively located between the transparent detection windows and between the lower detection window and the hollow metal frame bracket; the front plate of the stainless steel metal panel and the upper layer of the transparent detection window are provided with a through sample inlet and a sample outlet.

5. The glycated hemoglobin concentration detection device based on terahertz metamaterial according to claim 1, characterized in that: The data processing and analysis module includes a host computer, and the host computer is connected to the terahertz time-domain spectrometer.

6. The glycated hemoglobin concentration detection device based on terahertz metamaterial according to claim 1, characterized in that: The terahertz metamaterial chip is made of a resonant ring array etched by a planar gold film on a quartz glass substrate. A glycated hemoglobin-specific antibody is attached to the surface of the terahertz metamaterial chip to capture the glycated hemoglobin in a blood sample.

7. A method for detecting glycated hemoglobin concentration based on terahertz metamaterials, according to the device for detecting glycated hemoglobin concentration based on terahertz metamaterials according to any one of claims 1 to 6, characterized in that: The following steps are involved: (1) Prepare 9 samples of glycated hemoglobin with different concentration gradients. After loading, turn on the peristaltic pump switch and adjust the flow rate to 100 μL / min, and draw the sample into the terahertz metamaterial test module for antigen-antibody binding reaction for 20 min. Open the sample outlet of the terahertz metamaterial test module, adjust the flow rate of the peristaltic pump to 10 μL / min, and pass PBS buffer to clean the inner walls of all pipes and the inside of the terahertz metamaterial test module for 5 min. Finally, remove the PBS buffer from the sample inlet, and pump air into the peristaltic pump to remove the liquid on the chip surface in the terahertz metamaterial test module. Turn on the terahertz spectrometer for testing; (2) Analyzing and fitting the measured data by a host computer connected to the terahertz time-domain spectrometer, obtaining a relationship curve between the glycated hemoglobin concentration and the shift of the resonance peak, and obtaining a glycated hemoglobin standard concentration comparison card; (3) Add the collected blood from the diabetic patient into the sample container, repeat the step (1), correspond the amount of transmission peak shift to the previous comparison card, and obtain the glycated hemoglobin concentration in the patient's blood.

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