Raman spectrometer for antigen protein detection and use method thereof

Abstract

The invention discloses a Raman spectrometer for antigen protein detection and a use method of the Raman spectrometer. The Raman spectrometer comprises a sample table which moves along a straight line in the horizontal direction and a laser which is arranged above the sample table and emits laser towards the sample table, the use method comprises the following steps: placing the test strip dripped with a sample solution to be detected in the clamping groove of the sample table, and determining the initial position of the sample table; first data is input, the sample table is moved to a first position, and the first data is displacement of the sample table from the initial position to the first position; inputting second data, moving the sample table to a second position, the second data being the displacement of the sample table from the first position to the second position so as to ensure that the laser is focused on the to-be-detected position on the test strip; and starting a laser, carrying out Raman spectrum analysis on the test strip, and determining whether the sample to be detected contains the antigen protein or not. The Raman spectrometer provided by the invention can give consideration to both use portability and detection accuracy, and realizes rapid and accurate detection of the antigen protein.

Description

Technical Field

[0001] The present invention relates to the field of Raman spectrometers, and in particular to a Raman spectrometer for antigen protein detection and a use method thereof. Background Art

[0002] When laser is irradiated onto a sample, the scattered light whose propagation direction and frequency are changed is called Raman scattering. Since the frequency difference between the scattered light and the incident light, namely the Raman shift, has nothing to do with the frequency of the incident light, it is only related to the structure of the sample molecules themselves. Different chemical bonds or groups with different characteristics correspond to different Raman shifts, so the molecular structure of the sample can be determined by using Raman scattering through a Raman spectrometer.

[0003] Although traditional Raman spectrometers have high accuracy and sensitivity, the entire instrument structure is complex, has high requirements for the use site, and requires professional technicians to operate, so it cannot be used in scenarios that require rapid detection. In the existing technology, although handheld Raman spectrometers have broken through the limitations of sites and professional technicians, the detection errors caused by human factors are large and cannot meet the needs of daily detection.

[0004] Therefore, the prior art still needs to be improved and developed. Summary of the invention

[0005] In view of the above-mentioned deficiencies in the prior art, an object of the present invention is to provide a Raman spectrometer for antigen protein detection and a method of using the same, aiming to solve the problem that the existing Raman spectrometer cannot take into account both portability and detection accuracy.

[0006] The technical solution of the present invention is as follows:

[0007] In a first aspect of the present invention, a method for using a Raman spectrometer for antigen protein detection is provided, wherein the Raman spectrometer comprises a sample stage that moves in a straight line in a horizontal direction and a laser that is arranged above the sample stage and emits laser light toward the sample stage, and the method for using the Raman spectrometer comprises the steps of:

[0008] Placing a test strip dripped with a sample solution to be tested in a card slot of the sample stage, and determining an initial position of the sample stage;

[0009] Input first data to move the sample stage to a first position, wherein the first data is a displacement of the sample stage from the initial position to the first position;

[0010] Input second data to move the sample stage to a second position, wherein the second data is a displacement of the sample stage from the first position to the second position, so as to ensure that the laser is focused on the position to be tested on the test strip;

[0011] The laser is turned on to perform Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein.

[0012] In one embodiment, before the step of placing the test strip dripped with the sample solution to be tested in the card slot of the sample stage and determining the initial position of the sample stage, the steps include:

[0013] preparing nanoparticles with Raman reporter molecules and antibodies to antigenic proteins modified on the surface;

[0014] Connecting a sample pad, a conjugate pad, a nitrocellulose membrane and an absorption pad in sequence to prepare a test strip for antigen protein detection, wherein the conjugate pad is soaked with a solution of the nanoparticles;

[0015] The sample solution to be tested is dropped onto the sample pad to form the test paper strip with the sample solution to be tested.

[0016] In one embodiment, the step of turning on the laser and performing Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein specifically includes:

[0017] Adjusting the focal length of the laser so that the focus of the laser is on the test strip;

[0018] The position to be tested of the test strip is subjected to Raman spectroscopy analysis. If a characteristic peak of the Raman reporter molecule appears in the Raman spectrum of the position to be tested, it indicates that the sample to be tested contains antigen protein.

[0019] In one embodiment, the step of turning on the laser and performing Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein further includes:

[0020] Setting the laser to a single-shot test mode;

[0021] The laser performs a single sampling on the position to be tested of the test strip to obtain the Raman spectrum of the position to be tested. If the characteristic peak of the Raman reporter molecule appears in the Raman spectrum of the position to be tested, it indicates that the sample to be tested contains antigen protein.

[0022] In one embodiment, when the laser performs single sampling, it also includes:

[0023] The spectral signals around the position to be measured are sampled together and used as background signals, and the background signals are deducted from the Raman spectrum of the position to be measured.

[0024] In one embodiment, the step of turning on the laser and performing Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein further includes:

[0025] Setting the laser to a continuous test mode;

[0026] Input the third data to determine the initial sampling position;

[0027] Input fourth data to determine a sampling end position, wherein a test interval is formed between the sampling initial position and the sampling end position;

[0028] The sample stage moves within the test area, and at every preset time interval, the laser samples the test strip once, extracts the peak intensity point at the characteristic peak of the Raman reporter molecule, and draws a distance-intensity curve. If the shape of the distance-intensity curve matches the shape of the standard distance-intensity curve, it indicates that the sample to be tested contains antigen protein.

[0029] In one embodiment, the standard distance-intensity curve is a distance-intensity curve obtained by continuously testing a test strip dripped with an antigen protein solution and extracting the peak intensity point at the characteristic peak of the Raman reporter molecule.

[0030] In one embodiment, when the laser performs continuous sampling, it also includes:

[0031] The spectral signals around the interval to be measured are sampled together and used as background signals, and the background signals are deducted from the distance-intensity curve.

[0032] In one embodiment, after the step of turning on the laser and performing Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein, the method further includes:

[0033] The laser is turned off and the test strip is moved to the initial position.

[0034] The second aspect of the present invention provides a Raman spectrometer for antigen protein detection, and applies the above-mentioned method of use.

[0035] Beneficial effect: The present invention provides a Raman spectrometer for antigen protein detection and a method for using the same. The Raman spectrometer includes a sample stage that moves in a straight line in the horizontal direction and a laser that is arranged above the sample stage and emits laser light toward the sample stage. The method for using the Raman spectrometer includes the following steps: placing a test strip dripped with a sample solution to be tested in a card slot of the sample stage to determine the initial position of the sample stage; inputting first data to move the sample stage to a first position, wherein the first data is the displacement of the sample stage from the initial position to the first position; inputting second data to move the sample stage to a second position, wherein the second data is the displacement of the sample stage from the first position to the second position, so as to ensure that the laser is focused on the position to be tested on the test strip; turning on the laser to perform Raman spectroscopy analysis on the test strip to determine whether the sample to be tested carries antigen protein. The Raman spectrometer has a simple structure and can reduce errors caused by human operation, thereby taking into account both portability and detection accuracy, and realizing rapid and accurate detection of viral antigen proteins. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 Schematic diagram of a Raman spectrometer used for antigen protein detection in an embodiment of the present invention.

[0037] Figure 2 The figure is a flow chart of the method of using the Raman spectrometer for antigen protein detection in an embodiment of the present invention.

[0038] Figure 3 Schematic diagram of the change of characteristic peak intensity with integration time in an embodiment of the present invention.

[0039] Figure 4 Schematic diagram of the change of characteristic peak intensity with laser power in an embodiment of the present invention.

[0040] Figure 5 It is a schematic diagram of the change of the characteristic peak signal with the first position distance in the characterization results of the Raman spectrometer used for antigen protein detection in light conditions and dark conditions in an embodiment of the present invention.

[0041] Figure 6 Schematic diagram of the variation of characteristic peak intensity with the distance of the laser on the Z axis in an embodiment of the present invention.

[0042] Figure 7 1 is a standard distance-intensity curve at different concentrations when a material containing cyanate is used as a Raman reporter molecule in an embodiment of the present invention. DETAILED DESCRIPTION

[0043] The present invention provides a Raman spectrometer for antigen protein detection and a method for using the same. To make the purpose, technical solution and effect of the present invention clearer and more specific, the present invention is further described in detail below. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0044] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0045] Immunochromatography is a commonly used virus detection method. Generally, the antigen protein of the virus is used as the target detection object. After the sample to be tested is dripped on the immunochromatography detection plate, the target detection object is captured by the double antibody sandwich method, and then a preliminary judgment is made on the presence or absence of the antigen protein by the appearance of the test line and the control line on the immunochromatography detection plate, and then whether the virus is in the sample to be tested is judged. However, for the existing immunochromatography detection plate, the appearance of the test line and the control line can only be judged by the naked eye, and there is a deviation in naked eye observation, resulting in low accuracy of the test result. The present invention provides a Raman spectrometer for antigen protein detection and a method of using the same. The Raman spectrometer is used to detect a test strip dripped with a sample solution to be tested, and the Raman spectrum results at the test line and the control line are used to judge whether the sample to be tested contains the antigen protein, thereby eliminating the error of naked eye judgment. The Raman spectrometer has a simple structure and requires a small space, so that both portability and detection accuracy can be taken into account.

[0046] The present invention provides a Raman spectrometer for antigen protein detection, such as Figure 1 As shown, the Raman spectrometer includes a sample stage 200 that moves in a straight line in the horizontal direction and a laser 300 that is arranged above the sample stage 200 and emits laser light toward the sample stage 200. The moving direction of the sample stage 200 is set to the X-axis direction, that is, the sample stage can only move along the X-axis; the vertical direction is set to the Z-axis direction, that is, the laser 300 moves along the Z-axis, and the focal length of the laser emitted by the laser on the sample stage is adjusted by adjusting the distance between the laser 300 and the sample stage 200.

[0047] The method for using the Raman spectrometer for antigen protein detection of the present invention first includes the steps of:

[0048] S91, preparing nanoparticles with Raman reporter molecules and antibodies modified with antigenic proteins on the surface;

[0049] S92, sequentially connecting a sample pad, a conjugation pad, a nitrocellulose membrane and an absorption pad to prepare a test strip for antigen protein detection, wherein the conjugation pad is soaked with a solution of the nanoparticles;

[0050] S93, dropping the sample solution to be tested on the sample pad to form a test strip with the sample solution to be tested.

[0051] Specifically, the nanoparticles are gold nanoparticles or silver nanoparticles, and the antigen protein is an antigen protein of a virus, such as an antigen protein of a virus such as the new coronavirus, influenza A virus, influenza B virus, etc. The antigen protein is detected to determine whether the corresponding virus exists in the sample to be tested.

[0052] Specifically, a test line and a control line are also provided on the nitrocellulose membrane, wherein the test line is arranged close to the binding pad, and the control line is arranged close to the absorption pad. Specifically, a secondary antibody solution of the antibody of the antigen protein is drawn on the nitrocellulose membrane to form the control line; and a solution of another antibody of the antigen protein is drawn on the nitrocellulose membrane to form the test line. When the characteristic peak of the Raman reporter molecule appears in the Raman spectrum at the position of the test line, it indicates that the sample to be tested contains the antigen protein, thereby determining the presence of a specific virus in the sample to be tested.

[0053] After obtaining the test strip dripped with the sample solution to be tested, in the method for using the Raman spectrometer for antigen protein detection of the present invention, as follows: Figure 2 As shown, the steps include:

[0054] S100, placing the test strip dripped with the sample solution to be tested in the card slot of the sample stage, and determining the initial position of the sample stage;

[0055] S200, inputting first data to move the sample stage to a first position, wherein the first data is a displacement of the sample stage from the initial position to the first position;

[0056] S300, inputting second data to move the sample stage to a second position, wherein the second data is a displacement of the sample stage from the first position to the second position, so as to ensure that the laser is focused on the position to be tested on the test strip;

[0057] S400, turning on the laser, performing Raman spectroscopy analysis on the test strip, and determining whether the sample to be tested carries antigen protein.

[0058] Specifically, the card slot of the sample stage corresponds to the shape of the test strip, and placing the test strip in the card slot can ensure that the test strip moves synchronously with the sample stage, and the initial position is determined according to the size of the test strip. Optionally, when the laser emitted by the laser irradiates one end of the sample pad of the test strip, the position of the sample stage and the test strip at this time is set as the initial position, and the initial position is set as the origin of the X-axis, and the direction from the sample pad on the test strip to the absorption pad is the positive direction of the X-axis.

[0059] Specifically, the first data is a positive integer, and the first data is the displacement of the sample stage from the initial position to the first position; the second data is an integer, and the second data is the displacement of the sample stage from the first position to the second position. That is, according to the first data, the sample stage can only move in the positive direction of the X-axis and move to the first position whose distance relative to the initial position (origin) is the first data. Optionally, the first data is the distance between one end of the sample pad of the test strip and the position of the test line on the nitrocellulose membrane. When the second data is a positive integer, the sample stage is controlled to move in the positive direction of the X-axis; when the second data is a negative integer, the sample stage is controlled to move in the negative direction of the X-axis, and the distance between the second position and the first position is the absolute value of the second data. That is, the distance of the sample stage relative to the first position is adjusted by the second data. Optionally, the second position is a position within the position interval of 0-10mm on both sides of the first position. Optionally, the units of the first data and the second data are determined according to the accuracy of moving the sample stage, for example, the units of the first data and the second data can be millimeters (mm) or micrometers (μm).

[0060] Specifically, when the laser is turned on, the laser power and integration time (sampling time) of the laser have a great influence on the accuracy and efficiency of the spectrum testing process. Figure 3 and Figure 4 As shown, by increasing the integration time and laser power, the intensity of the characteristic peak can be increased, thereby ensuring the detection of low-concentration antigen proteins. However, when the laser power is too strong, the test strip may be burned, thereby affecting the final test result; when the integration time is too long, the noise caused by environmental factors increases, which is not conducive to testing and has low efficiency. Optionally, in the method of using the Raman spectrometer for antigen protein detection described in the present invention, the laser power of the laser is set to 30-50mW, and the integration time (sampling time) is 0.1-5s. Figure 5The figure shows different test results of the Raman spectrometer of the present invention under light conditions and dark conditions. Under the condition that other experimental conditions and parameters are consistent, due to the selection of appropriate laser power and sampling time, the characteristic peak signal in the Raman spectrum changes with the first position distance under light conditions and dark conditions. That is, the Raman spectrometer of the present invention has strong environmental adaptability, is not affected by natural light sources, and can be used in a portable manner. By specifically setting the laser power and sampling time of the laser, the standard test parameters of the conventional Raman test are guaranteed, the error of human operation is reduced, and the accuracy of the final test result is improved.

[0061] In one implementation, step S300 includes:

[0062] S310, adjusting the focal length of the laser so that the focus of the laser is on the test strip;

[0063] S320, performing Raman spectroscopy analysis on the position to be tested of the test strip. If a characteristic peak of the Raman reporter molecule appears in the Raman spectrum of the position to be tested, it indicates that the sample to be tested contains antigen protein.

[0064] The focal length of the laser is adjusted by adjusting the distance between the laser and the sample stage to find the optimal Raman signal intensity. Optionally, the focal point of the laser emission is ensured to be on the test line of the test strip. The position of the laser on the Z axis is set to adjust the focal length of the laser. Figure 6 As shown, when the distance of the laser on the Z axis changes, the characteristic peak intensity obtained by the final test also changes accordingly. When the distance is large, the laser is too far away from the sample stage, resulting in a defocused light spot, which affects the final test result; when the distance is too small, the laser is too close to the sample stage, resulting in excessive laser intensity and burning the test strip, causing the test result to shift, affecting the final test result. Optionally, the distance between the laser and the sample stage is adjusted to 1 cm. By adjusting the appropriate distance between the laser and the sample stage, the optimal Raman signal intensity can be obtained, and the test strip can be avoided from being burned due to excessive laser intensity, thereby achieving routine characterization, avoiding test result deviations caused by operator error, and improving the accuracy of the final test result.

[0065] In a specific implementation, the test line position of the test strip can be sampled once, thereby completing a single test of the test strip, which specifically includes the steps of:

[0066] S400, setting the laser to a single test mode;

[0067] S410, the laser samples the position to be tested of the test strip once to obtain the Raman spectrum of the position to be tested. If the characteristic peak of the Raman reporter molecule appears in the Raman spectrum of the position to be tested, it indicates that the sample to be tested contains antigen protein.

[0068] In one embodiment, when the laser performs single sampling, it also includes:

[0069] S420, sampling the spectral signals around the position to be measured and taking them as background signals, and subtracting the background signals from the Raman spectrum of the position to be measured.

[0070] Specifically, the signal intensity of the background signal is subtracted from the signal intensity of the Raman spectrum of the position to be tested. If the characteristic peak of the Raman reporter molecule exists in the processed Raman spectrum, the result is positive, and the antigen protein exists in the sample to be tested. Optionally, the quantitative detection of the antigen protein content in the sample to be tested can be achieved through the intensity of the characteristic peak and the characteristic peak intensity of the standard Raman reporter molecule in the Raman spectrum. Optionally, the Raman spectrum data of the standard Raman reporter molecule is stored in a database to identify the relationship between the sample to be tested and the standard substance by similarity matching. Subtracting the background signal from the Raman spectrum of the position to be tested can reduce the influence of the Raman signal molecules in the background, improve the accuracy of spectral detection, and avoid the occurrence of false positives.

[0071] Optionally, the sampling range needs to be preprocessed before a single test. Since the filter effect is not ideal when the laser wavelength is 785nm, the Raman signal close to 785nm is very easily affected by it, generating obvious background signals and the Raman signal is easy to exceed the range, so the Raman spectrum data needs to be clipped. Optionally, the range of the Raman spectrum is limited to 200cm -1 Up to 4000cm -1 , only the Raman scattering signals within this range are collected.

[0072] Optionally, the intensity range needs to be preprocessed before a single test. Since the interference between the imaging device and the external environment noise during the digitization and transmission process will affect the final digital image, the background noise can be reduced by adjusting the intensity range in the Raman spectroscopy characterization results.

[0073] In a specific implementation, a position interval of the test strip can be continuously sampled to complete the continuous test of the test strip, which specifically includes the steps of:

[0074] S500, setting the laser to a continuous test mode;

[0075] S510, input third data to determine the initial sampling position;

[0076] S520, input fourth data, determine the sampling end position, and form a test interval between the sampling initial position and the sampling end position;

[0077] S530, the sample stage moves within the test area, and at a preset time interval, the laser samples the test strip once, extracts the peak intensity point at the characteristic peak of the Raman reporter molecule, and makes a distance-intensity curve of the distance of the sampling point relative to the control line and the peak intensity of the characteristic peak. If the shape of the distance-intensity curve matches the shape of the standard distance-intensity curve, it indicates that the sample to be tested contains antigen protein.

[0078] Optionally, the third data and the fourth data are integers, and the third data is the displacement of the sample stage from the second position to the sampling initial position; the fourth data is the displacement of the sample stage from the second position to the sampling end position. When the third data and the fourth data are positive integers, the sample stage moves in the positive direction of the X-axis; when the third data and the fourth data are negative integers, the sample stage moves in the negative direction of the X-axis. That is, the Raman spectrometer continuously samples the test interval around the second position.

[0079] Optionally, the third data and the fourth data are both positive integers, and the third data is the displacement of the sample stage from the initial position to the sampling initial position; the fourth data is the displacement of the sample stage from the initial position to the sampling end position, and the initial position is the origin of the X-axis. That is, the Raman spectrometer directly samples a test interval of the test strip continuously.

[0080] Optionally, the sample stage moves at a uniform speed in the test interval, and at every preset time interval, the sample stage stops moving, allowing the laser to sample a certain point on the test strip until the laser completes the continuous test of the test strip. Optionally, by setting the test interval, the moving speed of the sample stage, and the number of test points for continuous testing, the Raman spectrometer automatically calculates the preset interval time to ensure that all points are fully sampled from one end of the test interval to the other. Through the fully automatic processing of the Raman spectrometer, the detection interval is continuously adjustable, so that the final output result of the spectrum is stable and reliable, effectively avoiding errors in manual operation and improving the accuracy of the final detection result.

[0081] In one embodiment, when the laser performs continuous sampling, it also includes:

[0082] S540: Sample the spectral signals around the interval to be measured and use them as background signals, and deduct the background signals from the distance-intensity curve.

[0083] Specifically, the signal intensity of the Raman spectrum of the test interval is subtracted from the signal intensity of the surrounding background signal. If the shape of the distance-intensity curve obtained after processing matches the shape of the standard distance-intensity curve, the result is positive and the antigen protein exists in the test sample. Figure 7 The standard distance-intensity curve at different concentrations when a material containing cyanate is used as a Raman reporter molecule is shown. When the shape of the curve obtained when the Raman spectrometer of the present invention performs continuous testing matches the standard distance-intensity curve, the result is positive, and the antigen protein is present in the sample to be tested. Optionally, the standard distance-intensity curve is a distance-intensity curve obtained by continuously testing a test strip dripped with an antigen protein solution and extracting the peak intensity point at the characteristic peak of the Raman reporter molecule. Optionally, the standard distance-intensity curve is saved in a database to identify the relationship between the sample to be tested and the standard substance by similarity matching. Subtracting the background signal from the Raman spectrum of the position to be tested can reduce the influence of the Raman signal molecules in the background, improve the accuracy of spectral detection, and avoid the occurrence of false positives.

[0084] Optionally, the sampling range needs to be preprocessed before continuous testing. Since the filter effect is not ideal when the laser wavelength is 785nm, the Raman signal close to 785nm is very easily affected by it, generating obvious background signals and the Raman signal is easy to exceed the range, so the Raman spectrum data needs to be clipped. Optionally, the range of the Raman spectrum is limited to 200cm -1 Up to 4000cm -1 , only the Raman scattering signals within this range are collected.

[0085] Optionally, the intensity range needs to be preprocessed before continuous testing. Since the interference between the imaging device and the external environmental noise during the digitization and transmission process will affect the final digital image, the background noise can be reduced by adjusting the intensity range in the Raman spectroscopy characterization results.

[0086] In one embodiment, after determining whether the sample to be tested contains antigenic protein, the method further comprises:

[0087] S600, turning off the laser and moving the test strip to the initial position.

[0088] By resetting the sample stage, the initial position of the next test is ensured to be the same as the initial position of the previous test, so as to reduce the detection error caused by position deviation, thereby improving the accuracy of the final detection result. Optionally, for test strips of different sizes, the initial position can be adjusted to ensure that the focus of the laser emitted by the laser is at one end of the test strip at the initial position.

[0089] Further, the Raman spectrometer for antigen protein detection provided by the embodiment of the present invention can achieve accurate detection of antigen protein by applying the method of use as described above. At the same time, the Raman spectrometer has a simple structure, and only needs to be provided with a sample stage that moves in the horizontal direction and a laser that is provided above the sample stage and emits a laser toward the sample stage, and has high portability. Therefore, the Raman spectrometer for antigen protein detection of the present invention can take into account both portability and detection accuracy, and realize rapid and accurate detection of viral antigen protein.

[0090] The present invention will be further described below by means of specific examples.

[0091] Example 1

[0092] Preparation of Nanoparticles:

[0093] Gold nanoparticles were prepared by sodium citrate hydrothermal reduction of chloroauric acid, Raman reporter molecules were covalently coupled to the gold nanoparticles, and then an antibody solution of a specific antigen protein was coated on the surface of the gold nanoparticles by electrostatic adsorption, thereby obtaining nanoparticles with surface modified Raman reporter molecules and antibodies of antigen proteins.

[0094] Example 2

[0095] Assembly of the test strips:

[0096] The sample pad and conjugate pad were immersed in the treatment solution for 1 h, then taken out and dried at 37°C.

[0097] The nanoparticle solution was dropped onto the conjugate pad, and the conjugate pad was dried at 37°C.

[0098] The sample pad was cut into a strip of 2.0 cm*4 mm, the binding pad was cut into a strip of 0.5 cm*4 mm, the nitrocellulose membrane was cut into a strip of 2.5 cm*4 mm, and the absorption pad was cut into a strip of 1.7 cm*4 mm. Another antibody solution of the specific antigen protein was drawn on 1 / 3 of the nitrocellulose membrane as a test line (T line), and a secondary antibody solution of the first antibody corresponding to the specific protein was drawn on 2 / 3 of the nitrocellulose membrane as a control line (C line). The nitrocellulose membrane was dried at 37°C.

[0099] Take a 6.0 cm*4 mm bottom plate, connect and fix the sample pad, the binding pad, the nitrocellulose membrane and the absorption pad on the bottom plate in sequence, and ensure that the test line of the nitrocellulose membrane is close to the sample pad, and the control line of the nitrocellulose membrane is close to the absorption pad, and drop the sample solution to be tested on the sample pad, so as to obtain the test strip with the sample solution to be tested of the present invention.

[0100] Example 3

[0101] Detection of antigenic protein:

[0102] In this embodiment, a reagent containing cyanate is used as a Raman reporter molecule. The characteristic vibration peak of this type of Raman reporter molecule is between 2000 and 2500 cm -1 Place the test strip in the card slot of the sample stage of the Raman spectrometer of the present invention, set the initial position so that the laser focus emitted by the laser is at one end of the sample pad of the test strip, and set the initial position to the X-axis origin. Enter the first data as 35mm, and move the sample stage to the first position at 35mm on the X-axis. After the sample stage is stationary, enter the second data as -1mm, move the sample stage to the second position at 34mm on the X-axis and 1mm away from the first position to ensure that the laser focus of the laser emission is on the test line of the test strip. Adjust the laser to 1cm on the Z axis, set the laser power to 50mW, the sampling time to 3s, turn on the laser and set the laser to single test mode to complete a single test. Set the laser to continuous test mode, enter the third data as -6mm, enter the fourth data as +6mm, and the number of sampling points is 50. First, move the sample stage to the sampling initial position of 28mm from the X axis and 6mm from the second position (test line), then move the sample stage to the sampling end position of 40mm from the X axis and 6mm from the second position (test line) at a uniform speed, and stop for 3s after each movement of 0.24mm, so that the laser can complete a sampling. After the entire sampling process is completed, obtain the test results of 50 points in the test interval from the sampling initial position to the sampling end position. After the test is completed, turn off the laser and move the sample stage to the initial position.

[0103] For single detection results, the signal intensity of the Raman spectrum at the position to be tested is subtracted from the signal intensity of the background signal. If the characteristic peak of cyanate is present in the processed Raman spectrum, the result is positive, and the antigen protein is present in the sample to be tested. For continuous detection results, the signal intensity of the Raman spectrum at the interval to be tested is subtracted from the signal intensity of the surrounding background signal. If the shape of the distance-intensity curve obtained after processing matches the shape of the standard distance-intensity curve of the cyanate Raman reporter molecule, the result is positive, and the antigen protein is present in the sample to be tested.

[0104] Example 4

[0105] like Figure 1The Raman spectrometer for antigen protein detection of the present invention is shown, including a base 100, a sample stage 200 and a laser 300. The base 100 includes a horizontally arranged substrate 110 and a slide rail 120 arranged vertically to the substrate 110. The sample stage 200 is horizontally arranged above the substrate 110, and a motor 210 is arranged between the sample stage 200 and the base 100, and the motor 210 controls the sample stage to move in a straight line direction. A slot 220 is also provided on the sample stage 200 to accommodate a test strip, and the laser 300 is arranged above the slot 220 to ensure that the laser focus emitted by the laser 300 is located on the test strip in the slot 220. Further, the laser 300 is movably connected to the slide rail 120, and the laser 300 can move along the slide rail 120 to adjust the distance between the laser 300 and the sample stage 200. By moving the sample stage 200, the laser 300 can sample a certain point of the test strip or a certain section of the test strip. The Raman spectrometer has a simple structure and can reduce errors caused by human operation, thereby taking into account both portability and detection accuracy, and realizing rapid and accurate detection of viral antigen proteins.

[0106] In summary, the present invention provides a Raman spectrometer for antigen protein detection and a method for using the same, wherein the Raman spectrometer includes a sample stage that moves in a straight line in the horizontal direction and a laser that is arranged above the sample stage and emits laser light toward the sample stage, and the method for using the same includes the steps of: placing a test strip dripped with a sample solution to be tested in a card slot of the sample stage, and determining the initial position of the sample stage; inputting first data to move the sample stage to a first position, wherein the first data is the displacement of the sample stage from the initial position to the first position; inputting second data to move the sample stage to a second position, wherein the second data is the displacement of the sample stage from the first position to the second position, so as to ensure that the laser is focused on the position to be tested on the test strip; turning on the laser, and performing Raman spectroscopy analysis on the test strip to determine whether the sample to be tested carries an antigen protein. The Raman spectrometer has a simple structure and can reduce errors caused by human operation, thereby taking into account both portability and detection accuracy, and realizing rapid and accurate detection of viral antigen proteins.

[0107] It should be understood that the application of the present invention is not limited to the above examples. For ordinary technicians in this field, improvements or changes can be made based on the above description. All these improvements and changes should fall within the scope of protection of the claims attached to the present invention.

Claims

1. A method for using a Raman spectrometer for antigen protein detection, It is characterized in that The Raman spectrometer comprises a sample stage that moves in a straight line in a horizontal direction and a laser that is arranged above the sample stage and emits laser light toward the sample stage. The method of using the laser spectrometer comprises the steps of: Placing a test strip dripped with a sample solution to be tested in a card slot of the sample stage, and determining an initial position of the sample stage; Input first data to move the sample stage to a first position, wherein the first data is a displacement of the sample stage from the initial position to the first position; Input second data to move the sample stage to a second position, wherein the second data is a displacement of the sample stage from the first position to the second position, so as to ensure that the laser is focused on the position to be tested on the test strip; The laser is turned on to perform Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein.

2. The method for using the Raman spectrometer for antigen protein detection according to claim 1, It is characterized in that Before the step of placing the test paper strip dripped with the sample solution to be tested in the card slot of the sample stage and determining the initial position of the sample stage, the method includes the following steps: preparing nanoparticles with Raman reporter molecules and antibodies to antigenic proteins modified on the surface; Connecting a sample pad, a conjugate pad, a nitrocellulose membrane and an absorption pad in sequence to prepare a test strip for antigen protein detection, wherein the conjugate pad is soaked with a solution of the nanoparticles; The sample solution to be tested is dropped onto the sample pad to form the test paper strip with the sample solution to be tested.

3. The method for using the Raman spectrometer for antigen protein detection according to claim 2, It is characterized in that In step 1, the laser is turned on to perform Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein, which specifically includes: Adjusting the focal length of the laser so that the focus of the laser is on the test strip; The position to be tested of the test strip is subjected to Raman spectroscopy analysis. If a characteristic peak of the Raman reporter molecule appears in the Raman spectrum of the position to be tested, it indicates that the sample to be tested contains antigen protein.

4. The method for using the Raman spectrometer for antigen protein detection according to claim 3, It is characterized in that The step of turning on the laser and performing Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein also includes: Setting the laser to a single-shot test mode; The laser performs a single sampling on the position to be tested of the test strip to obtain the Raman spectrum of the position to be tested. If the characteristic peak of the Raman reporter molecule appears in the Raman spectrum of the position to be tested, it indicates that the sample to be tested contains antigen protein.

5. The method for using the Raman spectrometer for antigen protein detection according to claim 4, It is characterized in that When the laser performs single sampling, it also includes: The spectral signals around the position to be measured are sampled together and used as background signals, and the background signals are deducted from the Raman spectrum of the position to be measured.

6. The method for using the Raman spectrometer for antigen protein detection according to claim 3, It is characterized in that The step of turning on the laser and performing Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein also includes: Setting the laser to a continuous test mode; Input the third data to determine the initial sampling position; Input fourth data to determine a sampling end position, wherein a test interval is formed between the sampling initial position and the sampling end position; The sample stage moves within the test area, and at a preset time interval, the laser samples the test strip once, extracts the peak intensity point at the characteristic peak of the Raman reporter molecule, and draws a distance-intensity curve. If the shape of the distance-intensity curve matches the shape of the standard distance-intensity curve, it indicates that the sample to be tested contains antigen protein.

7. The method for using the Raman spectrometer for antigen protein detection according to claim 6, It is characterized in that The standard distance-intensity curve is a distance-intensity curve obtained by continuously testing a test strip dripped with an antigen protein solution and extracting the peak intensity point at the characteristic peak of the Raman reporter molecule.

8. The method for using the Raman spectrometer for antigen protein detection according to any one of claims 6 to 7, It is characterized in that When the laser performs continuous sampling, it also includes: The spectral signals around the interval to be measured are sampled together and used as background signals, and the background signals are deducted from the distance-intensity curve.

9. The method for using the Raman spectrometer for antigen protein detection according to claim 1, It is characterized in that After the step of turning on the laser and performing Raman spectroscopy analysis on the test strip to determine whether the sample to be tested contains antigen protein, the method further includes: The laser is turned off and the test strip is moved to the initial position.

10. A Raman spectrometer for antigen protein detection, It is characterized in that The Raman spectrometer comprises a sample stage that moves in a horizontal direction and a laser that is arranged above the sample stage and emits laser light toward the sample stage, and is applied by the method of use as described in any one of claims 1 to 9.

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