A molybdenum disulfide thin-film sensor based on Raman detection and its manufacturing method

Abstract

This invention discloses a molybdenum disulfide thin-film sensor based on Raman detection and its manufacturing method, which can be used for the detection of tumor markers. The molybdenum disulfide thin-film silicon wafer is formed by layer-by-layer self-assembly of a molybdenum disulfide dispersion and a polydiallyl dimethyl ammonium chloride (PDDA) solution onto a clean silicon wafer after pretreatment with an argon plasma gun, forming a molybdenum disulfide thin film. The carboxyl groups provided by sodium cholate added during the ultrasonic activation of the molybdenum disulfide powder are utilized with EDC / NHS solution. Antibodies corresponding to tumor markers are then incubated, and after BSA blocking treatment, the corresponding tumor markers are captured and then Raman detection is performed.

Description

Technical Field

[0001] This invention relates to the field of sensor technology applicable to early tumor diagnosis, and in particular to a molybdenum disulfide thin film sensor based on Raman detection and its manufacturing method. Background Technology

[0002] Malignant tumors, with their high incidence and mortality rates, are characterized by short survival times, severe cancer pain, and significant adverse treatment reactions, seriously impacting patients' health and quality of life. Therefore, to achieve highly sensitive, rapid, and accurate detection of tumor markers, some novel detection methods have gradually transitioned from the laboratory stage to the clinical diagnostic stage, such as immunolabeling, magnetic bead labeling, magnetic bead-targeted drug delivery, electrochemical detection, and microfluidic technology. With the development of science and technology, advanced techniques and novel materials have been applied to the early detection of cancer. Among all these studies, Raman biosensors have attracted increasing interest from researchers.

[0003] Surface-enhanced Raman spectroscopy (SERS), a branch of spectroscopy, is based on the Raman scattering effect and has subsequently given rise to coherent Raman scattering and stimulated Raman scattering techniques. The Raman technology chips used are mainly divided into metal substrates and composite substrates.

[0004] Among them, metal substrates are the most widely used. Considering stability, metal substrates are mainly made of gold and silver. For example, in patent application CN 110530839 A, entitled "A Method for Preparing Molybdenum Disulfide / Silver Nano Immunoassay Substrate and Its Application in Repeatable Immunoassay", a method for preparing molybdenum disulfide / silver nano immunoassay substrate is disclosed.

[0005] When composite materials are used as substrates, nanomaterial substrates are often used. This is because substrates made by combining nanomaterials with metal nanotechnology can transmit stronger signals. At the same time, the cost of nanomaterials is much lower than that of metals such as gold and silver. More importantly, composite substrates reduce the signal interference problems caused by traditional metal substrates.

[0006] Nanomaterials, also known as ultrafine particulate materials, are composed of nanoparticles with particle sizes ranging from 1 to 100 nm. Their optical, thermal, electrical, magnetic, mechanical, and chemical properties differ significantly from those of bulk solids. Introducing nanomaterials into Raman detection leverages their superior mechanical, physical, and chemical properties to construct suitable substrates. For example, the excellent Raman enhancement effect of nanomaterials can significantly amplify the Raman signal intensity; their large specific surface area and good bioactivity allow for the addition of more antibodies, thereby improving sensor sensitivity; and their catalytic effects can accelerate reaction time. All these effects can improve sensor sensitivity, lower the detection limit, and shorten the detection response time.

[0007] Molybdenum disulfide (MoD), a graphene-like two-dimensional layered nanomaterial, has attracted widespread research interest due to its ultrathin structure, optical properties, and nanoscale characteristics. MoD possesses excellent biocompatibility, a large specific surface area, and is easily functionalized, allowing for direct contact with biomolecules and ensuring sample activity. This enables the effective loading of a large number of antibody molecules onto the MoD surface. Furthermore, MoD exhibits excellent Raman enhancement, significantly increasing Raman signal intensity. These properties make MoD highly suitable as a substrate for Raman biosensors. When a MoD substrate is coupled with a functionalized gold nanoparticle probe, the Raman scattering effect is further enhanced, improving the sensitivity and accuracy of the biosensor, lowering the detection limit, and showing broad application prospects in early tumor diagnosis.

[0008] For example, the patent application entitled "A Multi-Cavity Raman Substrate and Its Preparation Method and Application", with publication number CN111965160 A, discloses the following method for vertically growing molybdenum disulfide on the surface of pyramidal silicon: thiourea and sodium molybdate are dissolved in a mixed solution of water and alcohol, then transferred to a high-pressure reactor, pyramidal silicon is placed in the reactor, and then the reactor is placed in an oven for heating.

[0009] The existing preparation process is cumbersome. It involves allowing molybdenum disulfide to grow freely and vertically on a silicon wafer. The growth process is difficult to control, and the growth conditions are high temperature and high pressure, which poses certain safety risks and is not suitable for modern mass production. Summary of the Invention

[0010] The purpose of this invention is to address the shortcomings of existing technologies by providing a simple, low-cost biosensor for detecting tumor markers based on Raman detection of a molybdenum disulfide thin film, and a method for its fabrication that does not require high temperature and high pressure. To solve the above-mentioned technical problems, this invention provides the following technical solution: A method for manufacturing a molybdenum disulfide thin film sensor based on Raman detection, comprising a molybdenum disulfide thin film and a silicon wafer. The clean silicon wafer is pretreated with an ion gun and then sequentially immersed in a polydiallyldimethylammonium chloride solution and a molybdenum disulfide dispersion to form a molybdenum disulfide thin film on the silicon wafer. Antibodies are then immobilized on the molybdenum disulfide thin film using carboxyl groups.

[0011] Furthermore, the molybdenum disulfide film includes a PDDA layer and a molybdenum disulfide layer, which are alternately stacked; the ratio of the number of PDDA layers to the number of molybdenum disulfide layers is 1:1, and there are 5 PDDA layers.

[0012] Furthermore, the PDDA solution is obtained by mixing a 0.1% PDDA solution with a 1 mg / mL NaCl solution.

[0013] Furthermore, the molybdenum disulfide dispersion is obtained by mixing molybdenum disulfide powder and sodium cholate powder in deionized water and then sonicating the mixture using an ultrasonic probe.

[0014] Furthermore, the clean silicon wafer is obtained by ultrasonically cleaning the silicon wafer successively in acetone, isopropanol, and deionized water.

[0015] Furthermore, this includes the following steps:

[0016] Step 10: Cut the silicon wafer into appropriate sizes, and then ultrasonically clean the cut silicon wafer in acetone, isopropanol and deionized water for 10 minutes each. After that, blow the surface of the silicon wafer with nitrogen and then treat it with an argon plasma gun for 5 minutes.

[0017] Step 20: Place the pretreated silicon wafer into a 0.1% PDDA solution, soak for 10 minutes, remove it, rinse with deionized water, blow dry with nitrogen, and let it air dry completely.

[0018] Step 30: Place the silicon wafer into the molybdenum disulfide dispersion, soak for 10 minutes, remove it, wash it with deionized water and blow it dry with nitrogen, let it dry completely, and repeat step 20. Repeat this process 10 times to prepare a layer-by-layer self-assembled molybdenum disulfide film.

[0019] Step 40: Drop a solution of 1-(3-dimethylpropyl)-3-ethylcarbodiimide hydrochloride / N-hydroxythiosuccinimide onto the prepared molybdenum disulfide film for 2 hours to activate the carboxyl groups provided by sodium cholate in the molybdenum disulfide film. Then, wash and dry the molybdenum disulfide film with deionized water.

[0020] Step 50: Drop a 10% concentration onto the molybdenum disulfide film. -2 The corresponding antibodies for the tumor markers at a concentration of mg / mL were incubated in a refrigerator at 0-4℃ for 12 hours, then washed with deionized water and dried. Next, 1% bovine serum albumin solution was added and incubated for 1 hour to block the remaining active sites. Finally, the membrane was washed with deionized water and dried. Thus, the preparation of the molybdenum disulfide thin film sensor was completed.

[0021] Furthermore, in step 20, preparing the 0.1% PDDA solution by mass fraction includes the following steps:

[0022] Step 21: Dissolve 0.029 g of 35% PDDA solution in 10 mL of deionized water and mix well;

[0023] Step 22: Weigh 10 mg of NaCl and dissolve it in the PDDA solution obtained in step 201. Shake to mix it evenly to obtain a PDDA solution with a mass fraction of 0.1%.

[0024] Furthermore, in step 30, the preparation of the molybdenum disulfide dispersion includes the following steps:

[0025] Step 31: Weigh 2.625 g of molybdenum disulfide powder and 1.125 g of sodium cholate powder, and add them to 50 mL of deionized water and stir thoroughly.

[0026] Step 32: Place the ultrasonic probe of the cell disruptor in the solution, set the parameters of the cell disruptor to 750W power, 75% amplitude, and 1 second ultrasonic interval, and sonicate the solution for 12 hours. During the ultrasonic process, ensure that the solution temperature does not exceed 70℃. After the ultrasonic process is completed, let it stand and settle for 24 hours.

[0027] Step 33: After standing, take 30 mL of the supernatant solution into a centrifuge tube and centrifuge at 10000 Rpm for 30 minutes;

[0028] Step 34: After centrifugation, completely remove the supernatant, add 1 mL of deionized water to redissolve all the precipitate at the bottom of the centrifuge tube, and prepare a high-concentration molybdenum disulfide dispersion. Store it in a refrigerator at 0-6℃ for later use.

[0029] Furthermore, it also includes a method for preparing Raman marker probes, which involves taking 900 μL of gold nanoparticle solution in a test tube, adding 200 μL of 1 mM Raman marker, mixing thoroughly for 30 minutes, adding 10 μL of recognition antibody at a concentration of 1 mg / mL, mixing thoroughly for 2 hours, centrifuging at 7500 Rpm for 10 minutes, adding 100 μL of standard PBS solution, and storing in a refrigerator for later use;

[0030] The method for preparing the gold nanoparticle solution includes adding 400 μL of 50 mg / mL HAuCl4 solution to 200 mL of pure water, heating and stirring the solution to boiling, then adding 2.6 mL of 1% sodium citrate solution dropwise. After the solution turns wine red, continue heating and stirring for 15 minutes to ensure a complete reaction, thereby obtaining the gold nanoparticle solution.

[0031] Furthermore, in step 50, the 1-(3-dimethylpropyl)-3-ethylcarbodiimide hydrochloride / N-hydroxythiosuccinimide solution is prepared by using a standard phosphate buffer solution as a solvent, and is prepared from a 0.2M solution of the 1-(3-dimethylpropyl)-3-ethylcarbodiimide hydrochloride solution and a 0.02M solution of the N-hydroxythiosuccinimide solution.

[0032] The present invention also discloses a molybdenum disulfide thin film sensor based on Raman detection, which is prepared by the above-described method for manufacturing a molybdenum disulfide thin film sensor based on Raman detection.

[0033] Compared with the prior art, the beneficial effects of the present invention are:

[0034] 1. The molybdenum disulfide thin film sensor of this invention is a Raman biosensor. It captures corresponding tumor markers by modifying a molybdenum disulfide thin film substrate with a capture antibody. A recognition probe is formed by adding gold nanoparticles, Raman markers, and recognition antibodies. The recognition probe is labeled on the capture antibody that has successfully captured the tumor marker, and Raman detection is performed on the incubated molybdenum disulfide thin film. The preparation process of the molybdenum disulfide thin film in this invention is simple and low-cost, and the preparation process does not require high temperature and high pressure. After washing the silicon wafer, it is pretreated with a plasma gun and then directly immersed alternately in a molybdenum disulfide dispersion and a PDDA solution to quickly form a film. The molybdenum disulfide dispersion and PDDA solution can be reused multiple times and can be stored for use. The required silicon wafer area is extremely small, and molds can be designed for mass production to meet the needs of large-scale production.

[0035] 2. In this invention, the layer-by-layer self-assembly of molybdenum disulfide and PDDA enables the two-dimensional molybdenum disulfide to form a three-dimensional structure, greatly increasing the specific surface area and allowing more antibodies to attach to the substrate. Furthermore, the number of self-assembled layers of the molybdenum disulfide film is controllable. Additionally, the functionalized gold nanoparticle probes used exhibit Raman enhancement, significantly increasing the Raman signal intensity. Moreover, when the gold nanoparticles are labeled on the target, they couple with the molybdenum disulfide substrate, further enhancing the Raman signal. Both of these factors improve the sensor's sensitivity and lower the detection limit.

[0036] 3. Since both the molybdenum disulfide thin film and the silicon substrate used are conductive, this biosensor can also be used for electrochemical detection. Attached Figure Description

[0037] Figure 1 This is a schematic diagram of a molybdenum disulfide thin film sensor structure based on Raman detection according to the present invention;

[0038] Figure 2 This is a SEM characterization image of the molybdenum disulfide thin film substrate of the present invention;

[0039] Figure 3 This invention is for a concentration of 10 -6 -10 -13 Raman signal image of mg / mL AFP antigen using DTNB as a Raman marker;

[0040] Wherein: 1-molybdenum disulfide film, 2-silicon wafer. Detailed Implementation

[0041] To enhance understanding of the present invention, we will now describe it in further detail with reference to the accompanying drawings. These embodiments are for illustrative purposes only and do not constitute a limitation on the scope of protection of the present invention.

[0042] like Figure 1 As shown, a molybdenum disulfide thin film sensor based on Raman detection includes a molybdenum disulfide thin film 1 and a silicon wafer 2 as a substrate, wherein the molybdenum disulfide thin film is obtained by layer-by-layer self-assembly of molybdenum disulfide dispersion and PDDA solution.

[0043] In the molybdenum disulfide film 1, the ratio of the number of layers of molybdenum disulfide dispersion to the number of layers of PDDA solution is 1:1, and the final number of layers is 10.

[0044] The molybdenum disulfide dispersion was obtained by mixing molybdenum disulfide powder and sodium cholate powder in deionized water and then sonicating using the ultrasonic probe of a cell disruptor; the PDDA solution was obtained by mixing a 0.1% PDDA solution with a 1 mg / mL NaCl solution.

[0045] Before preparing the molybdenum disulfide film, silicon wafer 2 was ultrasonically cleaned with acetone, isopropanol, and deionized water for 10 minutes, and then pretreated with an argon plasma gun for 5 minutes.

[0046] The method for manufacturing the molybdenum disulfide thin-film sensor based on Raman detection in this invention includes the following steps:

[0047] Step 10: Cut silicon wafer 2 into appropriate sizes, and ultrasonically clean it for 10 minutes each with acetone, isopropanol and deionized water, blow the surface dry with nitrogen, and then treat it with argon plasma gun for 5 minutes.

[0048] Step 20: Place the pretreated silicon wafer 2 into a 0.1% PDDA solution, soak for 10 minutes, remove it, rinse with deionized water and dry with nitrogen, and let it air dry for 5 minutes.

[0049] Step 30: Place the silicon wafer 2 into the molybdenum disulfide dispersion, soak for 10 minutes, remove it, wash it with deionized water and blow it dry with nitrogen, let it sit for 5 minutes until it is completely dry, and repeat step 20. Repeat this process 10 times to prepare the layer-by-layer self-assembled molybdenum disulfide film 1.

[0050] Step 40: Take 900 μL of gold nanoparticle solution in a test tube, add 200 μL of 1 mM Raman marker, mix thoroughly for 30 minutes, then add 10 μL of 1 mg / mL recognition antibody, mix thoroughly for 2 hours, centrifuge at 7500 Rpm for 10 minutes, then add 100 μL of standard PBS solution, and store in the refrigerator for later use.

[0051] Step 50: Drop the 1-(3-dimethylpropyl)-3-ethylcarbodiimide hydrochloride / N-hydroxythiosuccinimide solution, i.e., EDC / NHS solution, onto the prepared molybdenum disulfide film for 2 hours to activate the carboxyl groups provided by sodium cholate in the molybdenum disulfide film, and then wash and dry with deionized water.

[0052] Step 60: Drop a 10% concentration onto the molybdenum disulfide film. -2 The corresponding antibodies for the tumor markers at a concentration of mg / mL were incubated in a refrigerator at 0-4℃ for 12 hours, then washed with deionized water and dried. Next, 1% bovine serum albumin solution was added and incubated for 1 hour to block the remaining active sites. Finally, the membrane was washed with deionized water and dried. Thus, the preparation of the molybdenum disulfide thin film sensor was completed.

[0053] The method for preparing the 0.1% PDDA solution in step 20 includes the following steps:

[0054] Step 21: Dissolve 0.029 g of 35% PDDA solution in 10 mL of deionized water and mix well;

[0055] Step 22: Weigh 10 mg of NaCl and dissolve it in the PDDA solution obtained in step 201. Shake to mix it evenly to obtain a PDDA solution with a mass fraction of 0.1%.

[0056] Preferably, the preparation method of the molybdenum disulfide dispersion in step 30 includes the following steps:

[0057] Step 31: Weigh 2.625 g of molybdenum disulfide powder and 1.125 g of sodium cholate powder, and add them to 50 mL of deionized water and stir thoroughly.

[0058] Step 32: Place the ultrasonic probe of the cell disruptor in the solution, set the parameters of the cell disruptor to 750W power, 75% amplitude, and 1 second ultrasonic interval, and sonicate the solution for 12 hours. During the ultrasonic process, ensure that the solution temperature does not exceed 70℃. After the ultrasonic process is completed, let it stand and settle for 24 hours.

[0059] Step 33: After standing, take 30 mL of the supernatant solution into a centrifuge tube and centrifuge at 10000 Rpm for 30 minutes;

[0060] Step 34: After centrifugation, completely remove the supernatant, add 1 mL of deionized water to redissolve all the precipitate at the bottom of the centrifuge tube, and prepare a high-concentration molybdenum disulfide dispersion. Store it in a refrigerator at 0-6℃ for later use.

[0061] The preparation method of the gold nanoparticle solution in step 40 is as follows: add 400 μL of HAuCl4 solution with a concentration of 50 mg / mL to 200 mL of pure water, heat and stir the solution until it boils, then add 2.6 mL of sodium citrate solution with a mass fraction of 1%, and after the solution turns wine red, continue to heat and stir for 15 minutes to ensure that the reaction is complete, and the gold nanoparticle solution can be obtained.

[0062] In step 50, the EDC / NHS solution is prepared using standard PBS solution as solvent, with EDC concentration of 0.2M and NHS concentration of 0.02M.

[0063] In this example, in order to construct a Raman biosensor targeting tumor markers, a layer-by-layer self-assembled molybdenum disulfide film 1 containing sodium cholate was modified on silicon wafer 2, and the desired antibody was immobilized on the molybdenum disulfide film using the carboxyl groups provided by sodium cholate.

[0064] Depend on Figure 3It is known that the molybdenum disulfide thin film prepared according to this method can detect AFP antigen, i.e., tumor marker, and the signal intensity decreases as the concentration of tumor marker decreases, that is, it has a relative response to different concentrations of tumor marker, and can be used for early diagnosis of tumor.

[0065] When a corresponding tumor marker is added, it is captured on the antibody due to specific recognition. Then, adding gold nanoparticle probes modified with Raman markers and recognizing antibodies allows for the recognition of the antibodies that have successfully captured the tumor markers. When different concentrations of tumor markers are added, the number of recognition probes remaining on the molybdenum disulfide film changes accordingly, and the corresponding Raman signal intensity also changes. Figure 2 As shown. Therefore, different concentrations of tumor markers will produce Raman signals of different intensities, thereby enabling the quantitative detection of tumor markers.

[0066] The above specific embodiments are only for illustrating the technical concept and structural features of the present invention, and are intended to enable those skilled in the art to implement them. However, the above content does not limit the scope of protection of the present invention. Any equivalent changes or modifications made in accordance with the spirit and essence of the present invention shall fall within the scope of protection of the present invention.

Claims

1. A method for manufacturing a molybdenum disulfide thin-film sensor based on Raman detection, comprising a molybdenum disulfide thin film and a silicon wafer, characterized in that: After being pretreated with an ion gun, the clean silicon wafer is sequentially immersed in a polydiallyldimethylammonium chloride solution and a molybdenum disulfide dispersion to form a molybdenum disulfide film on the silicon wafer. The antibody is then immobilized on the molybdenum disulfide film using carboxyl groups. Includes the following steps: Step 10: Cut the silicon wafer into appropriate sizes, and then ultrasonically clean the cut silicon wafer in acetone, isopropanol and deionized water for 10 minutes each. After that, blow the surface of the silicon wafer with nitrogen and then treat it with an argon plasma gun for 5 minutes. Step 20: Place the pretreated silicon wafer into a 0.1% PDDA solution, soak for 10 minutes, remove it, rinse with deionized water, blow dry with nitrogen, and let it air dry completely. The preparation of the 0.1% PDDA solution includes the following steps: Step 21: Dissolve 0.029g of 35% PDDA solution in 10mL of deionized water and mix well; Step 22: Weigh 10 mg of NaCl and dissolve it in the PDDA solution obtained in step 201. Shake to mix it evenly to obtain a PDDA solution with a mass fraction of 0.1%. Step 30: Put the silicon wafer into the molybdenum disulfide dispersion, soak it for 10 minutes, take it out, wash it with deionized water and blow it dry with nitrogen gas. Let it dry completely. Repeat step 20. Repeat this process 10 times to prepare a layer-by-layer self-assembled molybdenum disulfide film. The preparation of the molybdenum disulfide dispersion includes the following steps: Step 31: Weigh 2.625g of molybdenum disulfide powder and 1.125g of sodium cholate powder, and add them to 50mL of deionized water and stir thoroughly. Step 32: Place the ultrasonic probe of the cell disruptor in the solution, set the parameters of the cell disruptor to 750W power, 75% amplitude, and 1 second ultrasonic interval, and sonicate the solution for 12 hours. During the ultrasonic process, ensure that the solution temperature does not exceed 70℃. After the ultrasonication is completed, let it stand for 24 hours to settle. Step 33: After standing, take 30 mL of the supernatant solution into a centrifuge tube and centrifuge at 10000 Rpm for 30 minutes; Step 34: After centrifugation, completely remove the supernatant, then add 1 mL of deionized water to redissolve all the precipitate at the bottom of the centrifuge tube, preparing a high-concentration molybdenum disulfide dispersion, and store it in a refrigerator at 0-6℃ for later use; Step 40: Drop the 1-(3-dimethylpropyl)-3-ethylcarbodiimide hydrochloride / N-hydroxythiosuccinimide solution onto the prepared molybdenum disulfide film for 2 hours to activate the carboxyl groups provided by sodium cholate in the molybdenum disulfide film, and then wash and dry the molybdenum disulfide film with deionized water; Step 50: Drop a 10% concentration onto the molybdenum disulfide film. -2 The corresponding antibody for the tumor marker at a concentration of mg / mL was incubated in a refrigerator at 0-4℃ for 12 hours, then washed with deionized water and dried. A 1% bovine serum albumin solution was then added and incubated for 1 hour to block the remaining active sites. The membrane was then washed with deionized water and dried. Thus, the preparation of the molybdenum disulfide thin film sensor was completed.

2. The method for manufacturing a molybdenum disulfide thin-film sensor based on Raman detection according to claim 1, characterized in that: The molybdenum disulfide film includes a PDDA layer and a molybdenum disulfide layer, which are stacked alternately; the ratio of the number of PDDA layers to the number of molybdenum disulfide layers is 1:1, and there are 5 PDDA layers.

3. The method for manufacturing a molybdenum disulfide thin-film sensor based on Raman detection according to claim 2, characterized in that: The PDDA solution was obtained by mixing a 0.1% PDDA solution with a 1 mg / mL NaCl solution.

4. The method for manufacturing a molybdenum disulfide thin-film sensor based on Raman detection according to claim 3, characterized in that: The molybdenum disulfide dispersion is obtained by mixing molybdenum disulfide powder and sodium cholate powder in deionized water and then sonicating the mixture using an ultrasonic probe.

5. The method for manufacturing a molybdenum disulfide thin-film sensor based on Raman detection according to claim 1, characterized in that: The clean silicon wafers are obtained by ultrasonically cleaning the silicon wafers in acetone, isopropanol, and deionized water in sequence.

6. The method for manufacturing a molybdenum disulfide thin-film sensor based on Raman detection according to claim 1, characterized in that: It also includes a method for preparing Raman marker probes, which involves taking 900 μL of gold nanoparticle solution in a test tube, adding 200 μL of 1 mM Raman marker, mixing thoroughly for 30 minutes, adding 10 μL of recognition antibody at a concentration of 1 mg / mL, mixing thoroughly for 2 hours, centrifuging at 7500 Rpm for 10 minutes, adding 100 μL of standard PBS solution, and storing in a refrigerator for later use; The method for preparing the gold nanoparticle solution includes adding 400 μL of 50 mg / mL HAuCl4 solution to 200 mL of pure water, heating and stirring the solution to boiling, then adding 2.6 mL of 1% sodium citrate solution dropwise. After the solution turns wine red, continue heating and stirring for 15 minutes to ensure that the reaction is complete, thereby obtaining the gold nanoparticle solution.

7. The method for manufacturing a molybdenum disulfide thin-film sensor based on Raman detection according to claim 1, characterized in that: In step 40, the 1-(3-dimethylpropyl)-3-ethylcarbodiimide hydrochloride / N-hydroxythiosuccinimide solution is prepared by using a standard phosphate buffer solution as a solvent, and is prepared by using a 0.2M solution of the 1-(3-dimethylpropyl)-3-ethylcarbodiimide hydrochloride and a 0.02M solution of the N-hydroxythiosuccinimide.

8. A molybdenum disulfide thin-film sensor based on Raman detection, characterized in that: It is prepared by the manufacturing method of the molybdenum disulfide thin film sensor based on Raman detection as described in any one of claims 1-7.

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