A single-layer molybdenum disulfide phase sensor detection box for a novel coronavirus vaccine antigen protein and a detection method thereof

Abstract

The application relates to the field of rapid detection of new coronaviruses, in particular to a single-layer MoS2 phase sensor detection box for a new coronavirus vaccine antigen protein and a detection method thereof. The process that a MoS2 test sample is combined with a standard new coronavirus SARS-CoV-2 antibody is as follows: firstly, single-layer MoS2 is grown through a CVD method; further, ZnO is deposited on SiO2 / Si through atomic layer deposition (ALD); the single-layer MoS2 is transferred to a substrate through PMMA (polymethyl methacrylate) assisted wet transfer; the surface of the MoS2 is subjected to surface treatment; and the problem that current detection methods can only qualitatively detect whether there is a new coronavirus and cannot measure a specific concentration is solved. On the premise of ensuring high sensitivity, the actual concentration of a sample can be quantitatively measured. The pretreatment process of the sample is simple, the detection process is short in time consumption, the measurement result is intuitive and accurate, and the application has important significance for quantitative detection of the concentration of a new coronavirus for a large number of samples.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of rapid detection of new coronavirus, in particular to a single-layer MoS2 phase sensor detection box for new coronavirus vaccine antigen protein and a detection method thereof. BACKGROUND

[0002] Super surfaces based on high refractive index dielectric two-dimensional (2D) nanoparticles of subwavelength thickness provide an unprecedented platform for light field manipulation from the geometric Mie resonance effect. For example, a super lens is manufactured by using a dielectric 2D nanostructure with a thickness of λ / 10 to achieve phase modulation by sacrificing thickness. In the multi-layer MoS2, the phase accumulation caused by propagation and the interface phase shift make it possible for the resonance peak to have a 2π phase change. However, the high aspect ratio and low sidewall roughness of the dielectric resonant cavity bring great challenges to its manufacturing process. The development of chemical vapor deposition (CVD) method makes it possible to manufacture atomic 2D materials on a wafer scale integrated with current complementary metal oxide semiconductor (CMOS) technology. Zero reflection intensity provides a method to achieve singular behavior in phase control. When the amplitude of light tends to zero (zero or near zero), an extremely fast phase change (phase singularity) can be achieved near the critical point. Phase jump / gradient creates a singular optical field, including twisted photons and vortex optics, which provides a way for high-sensitivity biological analysis and virus detection. Although single-layer 2D materials are excellent visible light absorbers, 0.33nm thick graphene reaches 2.3%, and 0.7-0.8nm thick single-layer transition metal sulfide (MoS2, MoSe2 and WS2) reaches 5-10%, however, the absorption of up to 10% is still low for achieving perfect absorption, so it still needs special design to achieve zero light intensity. The present application is based on a Fabry-Perot cavity multi-layer film system of optical loss material / double transparent dielectric layer / Si substrate (i.e. single-layer MoS2 / ZnO / SiO2 / Si substrate), which realizes zero reflection (perfect absorption). And the extremely fast phase change that occurs at zero reflection is used to detect various concentrations of new coronavirus antigen N protein, and the mutated phase change makes the detection of new coronavirus antigen have a sensitivity of 10-12 M / L or even lower.

[0003] The detection steps of the traditional new coronavirus antigen colloidal gold method (latex method) are as follows:

[0004] Preparation before detection: 1. Check the validity period of the kit; 2. Place the sample to be tested (the sample to be tested is in a refrigerated or frozen state), test card, sample extraction solution and other test materials at room temperature; 3. Confirm the type of sample to be tested, which should be a respiratory sample (usually oropharyngeal, nasopharyngeal secretion); 4. Collect the sample, usually using the sampling swab provided with the kit; 5. Unscrew the sample extraction tube cap that has been filled with sample extraction solution; 6. Insert the sample collection swab into the sample extraction solution tube, rotate it against the tube wall for about 10 times (ensure that the swab head is in contact with the extraction solution), break the swab at the breakpoint, leave the swab head in the sample extraction solution tube, and tighten the extraction tube cap. Wait for 1 minute (try to configure a test tube rack and vertically fix the top of the extraction tube on the test tube rack).

[0005] Test detection: 1. Open the aluminum foil bag, take out the test card and place it flat with the sample addition hole facing up; 2. Unscrew the transparent cap on the top of the extraction tube, and add 3-4 drops (80uL) of the treated sample extract to the test card sample addition hole, and time it; 3. Observe the results, which should be observed within 15-20 minutes, and the results have no clinical significance after 30 minutes.

[0006] Observe the results: 1. When the test card quality control area (C area) has no red line, the test is invalid; 2. When the test card quality control area (C area) has a red line, and the test area (T area) has no red line, the test result should be determined as negative; 3. When the test card quality control area (C area) and the test area (T area) both have a red line, the test result should be determined as positive.

[0007] After testing: 1. Report the test results; 2. Dispose of medical waste generated on site, disinfect and package according to the new coronavirus medical waste treatment.

[0008] Traditional electrochemiluminescence immunoassay kit for detecting expression of antigen protein of new coronavirus vaccine:

[0009] The present application provides an electrochemiluminescence immunoassay kit for detecting expression of antigen protein of new coronavirus vaccine, which comprises at least a well plate coated with streptavidin, biotin-labeled anti-new coronavirus spike protein antibody 1, SULFO-labeled anti-new coronavirus spike protein antibody 2, washing solution, reading solution, new coronavirus S protein standard and new coronavirus RBD protein standard. The present application uses biotin-labeled anti-new coronavirus spike protein antibody 1 to connect with streptavidin plate as the stationary phase, and uses new coronavirus S protein and RBD protein as reference, which can be recognized by SULFO-labeled antibody 2, so as to detect the expression of new coronavirus antigen.

[0010] Since the molecule for detecting the new coronavirus in the colloidal gold method is unstable and easy to fall off, it needs to be fixed on the colloidal gold particles to enhance its stability, so the sensitivity is low. By using the method of physical adsorption, the antigen / antibody is easy to separate from the surface of the gold particles, the marker is unstable, and only qualitative or semi-quantitative results can be given, there are more false negative results, and it cannot meet the current detection needs of the severity of infection after the heavy Yang. In contrast, the present application takes advantage of the specific binding of antibodies and antigens, and uses a two-dimensional material multi-layer film system combined with ellipsometer optical detection, which can obtain quantitative results under the premise of ensuring high sensitivity, low cost and intuitive detection results, which is of great significance for assisting in judging the severity of new crown infection.

[0011] The latex method can be approximately considered that it uses latex particles instead of colloidal gold particles, and still can only give qualitative or semi-quantitative results, there are more false negative results, and both colloidal gold method and latex method need 15-20 minutes of color development time to obtain results. In contrast, the present application based on optical detection means can obtain detection results immediately, and the detection speed is faster. Compared with nucleic acid and antibody detection, the present application can detect in the early stage of new crown infection, and has lower cost, faster speed, higher stability and sensitivity, and is expected to obtain results within 10 minutes. SUMMARY

[0012] In order to solve the problems in the prior art, the main purpose of the present application is to provide a single-layer MoS2 phase sensor detection box for new coronavirus vaccine antigen protein and a detection method thereof, which can effectively solve the problems of the existing antigen detection kit in the background art, such as unstable marker, only qualitative or semi-quantitative results, more false negative results, inability to meet the current detection needs of the severity of infection after the heavy Yang, long waiting time for test results of 15-20 minutes, and no clinical significance of the results after 30 minutes.

[0013] To achieve the above purpose, the technical scheme adopted by the present application is as follows: a single-layer MoS2 phase sensor detection box for new coronavirus vaccine antigen protein, which is based on phase sensing technology and comprises a reagent box body, MoS2 test samples combined with standard new coronavirus SARS-CoV-2 antibodies, intrinsic MoS2 sample standard, a portable ellipsometer, an antigen buffer and a detection liquid.

[0014] The process of binding the MoS2 test sample to the standard new coronavirus SARS-CoV-2 antibody is as follows: first, monolayer MoS2 is grown by a CVD method, then ZnO is deposited on SiO2 / Si by atomic layer deposition (ALD) with a thickness of 34 nm, and monolayer MoS2 is transferred to the substrate by a polymethyl methacrylate (PMMA) assisted wet transfer method. Then the surface of MoS2 is treated to enhance the binding of the protein to be detected to MoS2.

[0015] The process of growing monolayer MoS2 by the CVD method is as follows: monolayer MoS2 is prepared by a molten salt method. First, an aqueous Na2MoO4 solution is prepared, and spin coating is performed on a SiO2 / Si substrate. Before growth, high-purity argon gas is introduced into a quartz tube furnace, and then the spin-coated substrate is placed in a ceramic boat located at the center of the tube furnace. S powder is placed in the ceramic boat 15-22 cm away from the upstream direction of Na2MoO4 in the furnace. After growing at 600-800℃ for 5-7 min, it is cooled to room temperature.

[0016] ZnO is deposited on SiO2 / Si by atomic layer deposition (ALD), and MoS2 flakes are transferred on a ZnO / SiO2 / Si substrate by a polymethyl methacrylate (PMMA) assisted wet transfer method. First, PMMA is spin-coated on the MoS2 flakes to obtain a uniform PMMA film. The sample is baked on a hot plate at 60-100℃ for 1-5 min, and after drying, the flakes are soaked in a supersaturated NaOH solution for 10-15 hours. Then the PMMA supported MoS2 flakes are separated from the original substrate and float on the solution. Finally, the floating film is transferred to the ZnO / SiO2 / Si substrate, and after the sample is dried, the PMMA is completely dissolved in acetone solution.

[0017] The process of surface treatment is as follows: first, the MoS2 flakes are soaked in an 11-mercapto-undecanoic acid (MUA) / ethanol solution at room temperature for 20-30 hours. The chemical linker MUA is attached to the surface of MoS2. The sulfur atoms and vacancies on the surface of MoS2 promote strong binding between the SH end of the MUA molecule and MoS2. The MUA molecules form a self-assembled monolayer on MoS2.

[0018] The MUA molecules are activated by soaking in a mixed solution of n-hydroxysuccinimide (NHS) and carbodiimide hydrochloride (EDC), and the MUA-treated MoS2 is soaked in a mixed ethanol solution of n-hydroxysuccinimide (NHS) and carbodiimide hydrochloride (EDC) for 6 hours at room temperature to activate the upper carboxyl group of the MUA molecule to form an n-hydroxysuccinimide ester. The active ester can easily react with the amino group provided by the antibody to form an amide bond. Then the SARS-CoV-2 antibody is fixed on the treated MoS2 sheet, and the antibody solution (600 mM) is soaked for 8-16 hours in a refrigerated environment.

[0019] The MUA solution is prepared as follows: 0.043672g MUA is dissolved in 2ml of ethanol, and the EDC mixed solution is prepared as follows: 0.023018g of NHS and 0.124192g of EDC are dissolved in 2ml of ethanol.

[0020] The buffer solution of the antigen is a carbonate buffer CB9.6 solution, which is prepared as follows: 1.59g of Na2CO3 and 2.94g of NaHCO3 are added to distilled water to make 1000mL, and stored at 2-6℃; the preparation of the phosphate buffer PBS solution of the antibody is as follows: 8.0g of NaCl, 0.2g of KCl, 1.44g of Na2HPO4, and 0.24g of KH2PO4 are dissolved in 800mL of distilled water, and the solution is adjusted to 7.4 with HCl, and finally distilled water is added to make 1L to obtain 0.01M PBS buffer solution. Different concentrations of 2019-nCoV antigen are soaked for 10 minutes, and then the MoS2 sheet is washed with deionized water and dried with nitrogen.

[0021] The present application can realize the following single-layer MoS2 phase sensor detection method for new coronavirus vaccine antigen protein (N protein), which comprises:

[0022] 1) Prepare different concentrations of new coronavirus N protein standard solution, and prepare N protein standard solution with concentrations of 10 -7 M / L, 10 -8 M / L, 10 - 9 M / L, 10 -10 M / L, 10 -11 M / L, 10 -12 M / L;

[0023] 2) The above new coronavirus N protein standard solution is added to the single-layer MoS2 sample after introducing functional molecules (including MUA, NHS / EDC and antibodies), respectively, and soaked for 10 minutes with different concentrations of 2019-nCoV antigen (N protein, purchased from Zhongdian Biotechnology Co., Ltd.), then the MoS2 flake is washed with deionized water and dried with nitrogen, and the optical signal value is read on the ellipsometer.

[0024] 3) A standard curve reflecting the relationship between the concentration of the new coronavirus N protein standard solution and the optical signal value is established;

[0025] 4) The new coronavirus N protein standard solution in step 2) is replaced with a new coronavirus vaccine antigen protein sample to be tested, and then the optical signal value of the sample to be tested is determined by the same method, and the concentration of the antigen protein in the sample to be tested is obtained by substituting the standard curve in step 3).

[0026] Compared with the prior art, the present application has at least the following advantages:

[0027] (I) The sample consumption of the method is low, and the method is stable. The kit can be used to detect the content of N protein in the new coronavirus, and quantitative results can be obtained, which is of great significance for assisting in judging the severity of new coronavirus infection.

[0028] (II) Compared with the colloidal gold method, the present application adopts an optical detection mode, which can obtain the detection result immediately, and significantly improves the detection speed and efficiency.

[0029] (III) Compared with nucleic acid and antibody detection, the present application can detect in the early stage of new coronavirus infection, and has lower cost, faster speed, higher stability and sensitivity, and is expected to obtain results within 10 minutes. DETAILED DESCRIPTION

[0030] The present application will be further described below according to the examples.

[0031] A single-layer MoS2 phase sensor detection box for new coronavirus vaccine antigen protein, the kit is based on phase sensing technology, comprising: a kit main body, a MoS2 test sample combined with a standard new coronavirus SARS-CoV-2 antibody, an intrinsic MoS2 sample standard, a portable ellipsometer, an antigen buffer and a detection liquid, wherein the portable ellipsometer is a ME-L-L type ellipsometer, and the phase difference between p-polarized light and s-polarized light of the sample before and after introducing functional molecules (including MUA, NHS / EDC and antibodies) is detected by the ellipsometer.

[0032] A single-layer MoS2 phase sensor detection kit for COVID-19 vaccine antigen proteins is characterized in that the portable ellipsometer is a ME-LL type ellipsometric spectrometer, which facilitates the detection of the phase difference between p-polarized and s-polarized light of samples before and after the introduction of functionalized molecules (including MUA, NHS / EDC and antibodies).

[0033] A method for detecting COVID-19 vaccine antigen proteins using a monolayer MoS2 phase sensor, characterized by comprising the following steps:

[0034] The first step is to prepare at least two different concentrations of SARS-CoV-2 N protein standard solutions;

[0035] The second step involves adding the above-mentioned SARS-CoV-2 N protein standard solution to the monolayer MoS2 sample after the introduction of functionalized molecules, immersing it in different concentrations of 2019-nCoV antigen, rinsing the MoS2 sheet with deionized water, gently drying it with nitrogen, and reading the optical signal value on an ellipsometry.

[0036] The third step is to establish a standard curve reflecting the relationship between the concentration of the SARS-CoV-2 N protein standard solution and the optical signal value;

[0037] The fourth step involves replacing the SARS-CoV-2 N protein standard solution from the second step with the SARS-CoV-2 vaccine antigen protein sample to be tested. Then, the optical signal value of the sample to be tested is measured using the same method and substituted into the standard curve from the third step to obtain the concentration of the antigen protein in the sample to be tested.

[0038] The process of binding MoS2 samples to standard SARS-CoV-2 antibodies was as follows: First, a monolayer of MoS2 was grown using CVD. Then, ZnO was deposited on SiO2 / Si with a thickness of 34 nm using atomic layer deposition (ALD), followed by polymethyl methacrylate (PMMA)-assisted wet transfer to transfer the monolayer MoS2 onto the substrate. Finally, the MoS2 surface was treated to enhance the binding of the target protein to MoS2.

[0039] The CVD method for growing monolayer MoS2 is as follows: Monolayer MoS2 is prepared using the molten salt method. First, a 5 mg / ml Na2MoO4 aqueous solution is prepared and spin-coated onto a SiO2 (300 nm) / Si substrate at 5000 rpm for 40 s. Before growth, high-purity argon gas is introduced into a quartz tube furnace at a rate of 80 sccm for 30 min. Next, the spin-coated substrate is placed in a ceramic boat located at the center of the tube furnace. 50 mg of sulfur powder is placed in the ceramic boat 20 cm upstream of the Na2MoO4 layer. After growth at 770 °C for 8 min, the substrate is cooled to room temperature for 120 min.

[0040] Further, ZnO was deposited on SiO2 / Si by atomic layer deposition (ALD) with a thickness of 34 nm, and MoS2 flakes were transferred on the ZnO / SiO2 / Si substrate by poly(methyl methacrylate) (PMMA)-assisted wet transfer. First, PMMA was spin-coated on the MoS2 flakes to obtain a uniform PMMA film. The sample was baked on a hot plate at 80 degrees for 2 minutes. After drying, the flakes were immersed in a supersaturated NaOH solution for 12 hours. Then the PMMA-supported MoS2 flakes were separated from the original substrate and floated on the solution. The floating film was then transferred to the ZnO / SiO2 / Si substrate, and after the sample was dried, the PMMA was completely dissolved with an acetone solution.

[0041] The surface treatment process is as follows: first, immerse the MoS2 flakes in an 11-mercapto-undecanoic acid (MUA) / ethanol solution (100 mM, purchased from Aladdin) at room temperature for 24 hours, attach the chemical linker MUA to the MoS2 surface, and the sulfur atoms and vacancies on the MoS2 surface promote strong binding between the SH end of the MUA molecule and the MoS2, forming a self-assembled monolayer of MUA molecules on the MoS2.

[0042] Further, the MUA molecules are activated by immersing an n-hydroxysuccinimide (NHS) and hydrochloric acid carbodiimide (EDC) mixed solution, and the MUA-treated MoS2 is immersed in an n-hydroxysuccinimide (NHS) and hydrochloric acid carbodiimide (EDC) mixed ethanol solution at room temperature for 6 hours (purchased from Aladdin), to activate the upper end carboxyl group of the MUA molecule to form an n-hydroxysuccinimide ester. This active ester can easily react with the amino group provided by the antibody to form an amide bond. Then the SARS-CoV-2 antibody is fixed on the treated MoS2 flakes, and the antibody solution (600 mM) is immersed in a refrigerated environment for 12 hours.

[0043] In the present application, the MUA solution is prepared as follows: 0.043672 g of MUA is dissolved in 2 ml of ethanol, and the NHS and EDC mixed solution is prepared as follows: 0.023018 g of NHS and 0.124192 g of EDC are dissolved in 2 ml of ethanol.

[0044] In the present application, the carbonate buffer solution CB9.6 solution of the antigen is prepared as follows: 1.59 g of Na2CO3 and 2.94 g of NaHCO3 are added to distilled water to make up to 1000 mL, and stored at 4℃. The preparation of the phosphate buffer solution PBS solution of the antibody is as follows: 8.0 g of NaCl, 0.2 g of KCl, 1.44 g of Na2HPO4, and 0.24 g of KH2PO4 are dissolved in 800 mL of distilled water, the solution is adjusted to 7.4 with HCl, and finally distilled water is added to make up to 1 L to obtain 0.01 M PBS buffer solution.

[0045] Further, different concentrations of 2019-nCoV antigen (purchased from Zhongdian Biotechnology Co., Ltd.) are introduced to soak for 10 minutes, then the MoS2 sheet is rinsed with deionized water and gently dried with nitrogen.

[0046] In the kit of the present application, the detection principle of the kit of the present application is that the adsorption of 2019-nCoV antigen (NCPN2) changes the stacking environment based on the atomic-level MoS2 sample, thereby changing the wavelength position of the phase mutation. After introducing different concentrations of 2019-nCoV antigen (NCPN2), different concentrations of antigen will cause the wavelength of phase mutation to be blue-shifted, and the wavelength shift has linear sensitivity to the concentration of novel coronavirus antigen. Based on the single-layer MoS2 phase sensor, the novel coronavirus antigen with extremely low concentration can be clearly detected.

[0047] Compared with the prior art, the present application has at least the following advantages:

[0048] (I) The present method has low sample consumption and is stable. The kit can be used to detect the content of N protein in the novel coronavirus, and quantitative results can be obtained, which is of great significance for assisting in judging the severity of novel coronavirus infection.

[0049] (II) Compared with the colloidal gold method, the present application adopts an optical detection mode, and the detection result can be obtained immediately, significantly improving the detection speed and efficiency.

[0050] (III) Compared with nucleic acid and antibody detection, the present application can detect in the early stage of novel coronavirus infection, and has lower cost, faster speed, higher stability and sensitivity, and the result can be obtained within 10 minutes.

Claims

1. A single-layer MoS2 phase sensor detection box for a novel coronavirus vaccine antigen protein, characterized in that, The detection box comprises a kit main body, a MoS2 test sample combined with a standard new coronavirus SARS-CoV-2 antibody, an intrinsic MoS2 sample standard, an antigen buffer and a detection liquid, and a portable ellipsometer is further arranged in the detection box, the MoS2 test sample combined with the standard new coronavirus SARS-CoV-2 antibody is first grown by a CVD method to form a single-layer MoS2, ZnO is further deposited on SiO2 / Si by atomic layer deposition (ALD), and the single-layer MoS2 is transferred to a ZnO / SiO2 / Si substrate by a polymethyl methacrylate (PMMA) assisted wet transfer method, and then the surface of the MoS2 is treated; Different concentrations of 2019-nCoV antigens are introduced for soaking, and then the MoS2 sheet is washed with deionized water and dried with nitrogen, and the optical signal value is read on an ellipsometer. The process of growing the single-layer MoS2 by the CVD method comprises the following steps: single-layer MoS2 is prepared by a molten salt method, first, a Na2MoO4 aqueous solution is prepared, spin coating is performed on a SiO2 / Si substrate, before growth, high-purity argon gas is introduced into a quartz tube furnace, then the spin-coated substrate is placed in a ceramic boat located at the center of the tube furnace, S powder is placed in the ceramic boat 15-22 cm away from the upstream direction of the Na2MoO4, and after growth at 600-800 DEG C for 5-7 min, the temperature is cooled to room temperature.

2. The single-layer MoS2 phase sensor detection box of a new coronavirus vaccine antigen protein according to claim 1, characterized in that The portable ellipsometer is an ME-L-L type ellipsometer, which is convenient for detecting the phase difference of p-polarized light and s-polarized light of the sample before and after the functional molecules are introduced.

3. The single-layer MoS2 phase sensor detection box of a new coronavirus vaccine antigen protein according to claim 1, characterized in that ZnO is deposited on SiO2 / Si by atomic layer deposition (ALD), which is used for transferring the MoS2 sheet to the ZnO / SiO2 / Si substrate by a polymethyl methacrylate (PMMA) assisted wet transfer method; first, PMMA is spin-coated on the MoS2 sheet to obtain a uniform PMMA film, the sample is baked on a hot plate at 60-100 DEG C for 1-5 min, after drying, the sheet is soaked in a supersaturated NaOH solution for 10-15 hours, then the PMMA supported MoS2 sheet is separated from the original substrate and floats on the solution, and then the floating film is transferred to the ZnO / SiO2 / Si substrate, and after the sample is dried, the PMMA is completely dissolved in acetone solution; The process of surface treatment comprises the following steps: first, the MoS2 sheet is soaked in an 11-mercapto-undecanoic acid (MUA) / ethanol solution at room temperature for 20-30 hours, the chemical linker MUA is attached to the surface of the MoS2, the sulfur atoms and vacancies on the surface of the MoS2 promote the strong binding between the SH end of the MUA molecule and the MoS2, and the MUA molecules form a self-assembled monolayer on the MoS2. The MUA molecules are activated by soaking in a mixed solution of n-hydroxysuccinimide (NHS) and ethyl carbodiimide hydrochloride (EDC). The MUA-treated MoS2 is soaked in a mixed ethanol solution of n-hydroxysuccinimide (NHS) and ethyl carbodiimide hydrochloride (EDC) at room temperature for 6 hours to activate the upper carboxyl group of the MUA molecules to form n-hydroxysuccinimide ester, which reacts with the amino group provided by the antibody to form an amide bond. Then the SARS-CoV-2 antibody is fixed on the treated MoS2 sheet and soaked in a 600 mM antibody solution in a refrigerated environment for 8-16 hours.

4. The single-layer MoS2 phase sensor detection box of a new coronavirus vaccine antigen protein according to claim 3, characterized in that The MUA solution is prepared as follows: 0.043672 g of MUA is dissolved in 2 ml of ethanol. The EDC mixed solution is prepared as follows: 0.023018 g of NHS and 0.124192 g of EDC are dissolved in 2 ml of ethanol. The buffer solution for the antigen is a carbonate buffer CB9.6 solution, which is prepared as follows: 1.59 g of Na2CO3 and 2.94 g of NaHCO3 are added to distilled water to make 1000 mL, and stored at 2-6°C. The phosphate buffer PBS solution for the antibody is prepared as follows: 8.0 g of NaCl, 0.2 g of KCl, 1.44 g of Na2HPO4, and 0.24 g of KH2PO4 are dissolved in 800 mL of distilled water, and the solution is adjusted to 7.4 with HCl. Finally, distilled water is added to make 1 L to obtain a 0.01 M PBS buffer solution. Different concentrations of 2019-nCoV antigen are soaked for 10 minutes, followed by rinsing the MoS2 sheet with deionized water and gently drying with nitrogen.

5. The single-layer MoS2 phase sensor detection box of a new coronavirus vaccine antigen protein according to claim 2, characterized in that The functional molecules include MUA, NHS / EDC, and antibodies. The MUA molecules are activated by soaking in a mixed solution of n-hydroxysuccinimide (NHS) and ethyl carbodiimide hydrochloride (EDC). The MUA-treated MoS2 is soaked in a mixed ethanol solution of n-hydroxysuccinimide (NHS) and ethyl carbodiimide hydrochloride (EDC) at room temperature for 6 hours to activate the upper carboxyl group of the MUA molecules to form n-hydroxysuccinimide ester, which reacts with the amino group provided by the antibody to form an amide bond. Then the SARS-CoV-2 antibody is fixed on the treated MoS2 sheet and soaked in a 600 mM antibody solution in a refrigerated environment for 8-16 hours. The MUA solution is prepared as follows: 0.043672 g of MUA is dissolved in 2 ml of ethanol. The EDC mixed solution is prepared as follows: 0.023018 g of NHS and 0.124192 g of EDC are dissolved in 2 ml of ethanol. The buffer solution for the antigen is a carbonate buffer CB9.6 solution, which is prepared as follows: 1.59 g of Na2CO3 and 2.94 g of NaHCO3 are added to distilled water to make 1000 mL, and stored at 2-6°C. The phosphate buffer PBS solution for the antibody is prepared as follows: 8.0 g of NaCl, 0.2 g of KCl, 1.44 g of Na2HPO4, and 0.24 g of KH2PO4 are dissolved in 800 mL of distilled water, and the solution is adjusted to 7.4 with HCl. Finally, distilled water is added to make 1 L to obtain a 0.01 M PBS buffer solution. Different concentrations of 2019-nCoV antigen are soaked for 10 minutes, followed by rinsing the MoS2 sheet with deionized water and gently drying with nitrogen. The functional molecules include MUA, NHS / EDC, and antibodies.