An aerosol viral particle detection device and method

Abstract

The application discloses an aerosol virus particle detection device, which comprises an aerosol collection module, a microfluidic chip module, a fluorescence detection module and a control system. The microfluidic chip module comprises a first substrate, a detection chamber and a biochip. The other end of the flow guide pipe is communicated with the detection chamber. The biochip comprises a second substrate and a nano array structure. The nano array structure comprises a plurality of first nano columns and a plurality of nano flow guide plates. The plurality of first nano columns and the plurality of nano flow guide plates are alternately arranged in multiple rows. The spacing between adjacent first nano columns and the spacing between adjacent nano flow guide plates in each row gradually decrease along the direction of virus suspension flow. The surface of the first nano column is modified with target virus specific capture antibodies. The application also discloses an aerosol virus particle detection method. The application integrates virus particle transportation, interception and dispersion into one, realizes sufficient combination of virus particles and antibodies, and finally realizes accurate detection of single virus particles through fluorescence labeling.

Description

Technical Field

[0001] This invention relates to the field of virus detection technology, and in particular to an aerosol virus particle detection device and method. Background Technology

[0002] Aerosols are the primary carriers of viruses transmitted through the air. Their particle size typically ranges from 0.01 to 10 μm, and they are characterized by wide diffusion range and long survival time, posing a serious threat to public health. Therefore, achieving efficient collection and accurate detection of virus-carrying aerosol particles in the air is a key technological support for blocking virus transmission and carrying out early prevention and control.

[0003] Currently, existing aerosol virus detection technologies suffer from the following core defects, making it difficult to meet actual prevention and control needs: low detection sensitivity, difficulty in detecting single viruses, and the process requires specialized laboratory equipment, is cumbersome, and has a detection cycle of 1-4 hours, failing to meet the needs of rapid on-site screening; in some microfluidic detection devices, the virus and antibody do not bind sufficiently, resulting in poor detection accuracy; low device integration, high operational threshold, and existing detection systems are mostly decentralized structures, requiring independent equipment for sampling, sample processing, and detection, which are bulky, inconvenient to carry, and require professional technicians to operate, making them unsuitable for on-site monitoring scenarios such as airports, subway stations, and hospitals. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the present invention aims to provide an aerosol virus particle detection device and method.

[0005] To achieve the above objectives, an embodiment of the present invention provides the following technical solution:

[0006] An aerosol virus particle detection device, comprising:

[0007] An aerosol collection module includes a gas-liquid mixing chamber, a storage bottle, an air inlet pipe, a liquid inlet pipe, and a guide pipe. One end of the air inlet pipe extends into the gas-liquid mixing chamber and is inclined. Both ends of the liquid inlet pipe are connected to the gas-liquid mixing chamber and the storage bottle, respectively. One end of the guide pipe is connected to the bottom of the gas-liquid mixing chamber.

[0008] A microfluidic chip module includes a first substrate, a detection chamber, and a biochip disposed within the detection chamber. The other end of a flow guide tube is connected to the detection chamber. The biochip includes a second substrate and a nanoarray structure disposed on the second substrate. The nanoarray structure includes multiple first nanopillars and multiple nanoflow guides. The multiple first nanopillars and multiple nanoflow guides are arranged in multiple rows and alternately. The spacing between adjacent first nanopillars and the spacing between adjacent nanoflow guides in each row gradually decreases along the direction of virus suspension flow. The surface of the first nanopillars is modified with a target virus-specific capture antibody.

[0009] A fluorescence detection module, located corresponding to the detection chamber, is used to capture, process, and count fluorescence signals;

[0010] The control system is used to control the coordinated operation of the aerosol collection module, the microfluidic chip module, and the fluorescence detection module.

[0011] As a further improvement of the present invention, the inclination angle of the air inlet pipe is 30°-45°, one end of the air inlet pipe is 0.8cm-1.2cm away from the inner wall of the gas-liquid mixing chamber, and one end of the air inlet pipe is 2.5cm-3.5cm away from the liquid surface of the gas-liquid mixing chamber.

[0012] As a further improvement of the present invention, the biochip is detachably installed in the detection chamber.

[0013] As a further improvement of the present invention, the microfluidic chip module further includes an inlet and a delivery channel. The inlet is connected to the other end of the guide tube, and the two ends of the delivery channel are respectively connected to the inlet and the detection chamber. A rinsing liquid bottle and a fluorescent labeling liquid bottle are also provided, and the rinsing liquid bottle and the fluorescent labeling liquid bottle are both connected to the guide tube.

[0014] As a further improvement of the present invention, the height of the first nanopillar and the nanofluid plate is 200nm-400nm, the diameter of the first nanopillar is 80nm-120nm, the thickness of the nanofluid plate is 80nm-120nm, and the spacing between adjacent first nanopillars is 200nm-300nm.

[0015] As a further improvement of the present invention, the adjacent nano-guide plates in each row form a figure-eight shape.

[0016] As a further improvement of the present invention, a second nanopillar is provided between at least one adjacent nano-guide plate in each row.

[0017] As a further improvement of the present invention, the fluorescence detection module includes a fluorescence excitation unit, an optical imaging unit, and a signal processing unit. The fluorescence excitation unit includes a laser and a focusing lens. The optical imaging unit includes a filter group, a fluorescence microscope, and a CCD camera. The signal processing unit includes an image acquisition card and a data processing module.

[0018] As a further improvement of the present invention, the control system includes a core controller, a sensor group, a human-machine interaction module, and a data storage module. The core controller is a microcontroller or a programmable logic controller. The sensor group includes a gas flow meter and a liquid flow meter. The human-machine interaction module includes a touch screen and status indicator lights.

[0019] A method for detecting aerosol virus particles, using the aforementioned aerosol virus particle detection device, includes the following steps:

[0020] (1) Aerosol collection: The buffer solution is injected into the gas-liquid mixing chamber from the storage bottle. The control system is started. Air is rushed into the gas-liquid mixing chamber at an angle through the air inlet pipe. It forms a gas-liquid impact mixing with the injected buffer solution. Virus particles in the aerosol are captured into the buffer solution to form a virus suspension.

[0021] (2) Virus suspension delivery: The virus suspension is delivered from the gas-liquid mixing chamber to the detection chamber of the microfluidic chip module;

[0022] (3) Virus capture: The virus suspension comes into contact with the nanoarray structure of the biochip in the detection chamber. The nanofluid plate diverts the virus particles. The modified target virus-specific capture antibody on the surface of the first nanopillar binds specifically to the first antigen on the surface of the virus particles to complete the virus capture.

[0023] (4) Fluorescent labeling: Fluorescent labeling solution is delivered to the detection chamber. The fluorescent labeling detection antibody in the fluorescent labeling solution binds to the second antigen corresponding to the surface of the virus particles, and the captured virus is fluorescently labeled.

[0024] (5) Fluorescence detection: The fluorescence detection module is activated to capture the fluorescence signal of the nanoarray structure, process the fluorescence signal and count the fluorescence spots;

[0025] (6) Result output: The control system outputs the number or concentration of virus particles based on the fluorescence dot count results to complete the detection.

[0026] The beneficial effects of this invention are:

[0027] This invention achieves efficient collection of aerosol viruses through gas-liquid impact mixing. Combined with a biochip nanoarray structure, the nanoarray disperses virus particles through gaps between multiple nano-guide plates, preventing virus aggregation while ensuring sufficient contact with the first and second nanopillars for interception. A first-target virus-specific antibody captures the virus, achieving full binding and capture of virus particles. The virus is labeled with fluorescently modified antibodies, and a fluorescence detection module enables accurate detection of individual viruses. This invention has a detection limit as low as a single virus particle, a detection time of ≤30 minutes, high integration, and simple operation, making it widely applicable for aerosol virus monitoring and disease control in public places, meeting the practical needs of public health prevention and control. Attached Figure Description

[0028] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0029] Figure 1 This is a schematic diagram of a preferred embodiment of the present invention;

[0030] Figure 2 This is an exploded structural diagram of a microfluidic chip module according to a preferred embodiment of the present invention.

[0031] Figure 3 This is a partial structural schematic diagram of a biochip according to a preferred embodiment of the present invention;

[0032] Figure 4 The block diagram of the control system of the preferred embodiment of the present invention is shown, which is connected to the aerosol collection module, the microfluidic chip module and the fluorescence detection module respectively.

[0033] In the diagram: 1. Aerosol collection module; 11. Gas-liquid mixing chamber; 111. Exhaust port; 12. Storage bottle; 13. Air inlet pipe; 130. Aerosol sampling head; 131. Air pump; 132. Airflow valve; 14. Liquid inlet pipe; 141. First peristaltic pump; 15. Guide pipe; 151. Second peristaltic pump; 16. Rinse solution bottle; 161. First branch pipe; 162. Third peristaltic pump; 17. Fluorescent labeling solution bottle; 171. Second branch pipe; 172. Fourth peristaltic pump; 2. Microfluidic chip module; 21. First substrate; 211. Groove; 212. Elastic silicone cap; 22. Detection chamber; 23. Biochip; 231. Second substrate; 232. First nanopillar; 233. Nanoparticle guide plate. 234. Virus suspension flow direction; 235. Second nanocolumn; 24. Inlet; 25. Delivery channel; 26. Drain; 27. Collection bottle; 28. Cover plate; 291. Virus particles; 292. Fluorescently labeled detection antibody; 293. Fluorescent label; 3. Fluorescent detection module; 31. Fluorescent excitation unit; 311. Laser; 312. Focusing lens; 313. Excitation light; 32. Optical imaging unit; 321. Beam splitter; 322. Fluorescence microscope; 323. CCD camera; 324. Excitation filter; 325. Fluorescent filter; 33. Signal processing unit; 4. Control system; 41. Core controller; 42. Sensor group; 43. Human-computer interaction module; 44. Data storage module. Detailed Implementation

[0034] To enable those skilled in the art to better understand the technical solutions of this invention, the technical solutions of the embodiments of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of this invention.

[0035] Please see Figures 1-3This application discloses an aerosol virus particle detection device, comprising: an aerosol collection module 1, a microfluidic chip module 2, a fluorescence detection module 3, and a control system 4. The aerosol collection module 1 includes a gas-liquid mixing chamber 11, a liquid storage bottle 12, an air inlet pipe 13, a liquid inlet pipe 14, and a guide pipe 15. One end of the air inlet pipe 13 extends into the gas-liquid mixing chamber 11 and is inclined. The two ends of the liquid inlet pipe 14 are respectively connected to the gas-liquid mixing chamber 11 and the liquid storage bottle 12. One end of the guide pipe 15 is connected to the bottom of the gas-liquid mixing chamber 11. The microfluidic chip module 2 includes a first substrate 21, a detection chamber 22, and a biochip 23 disposed within the detection chamber 22. The other end of the flow guide 15 is connected to the detection chamber 22. The biochip 23 includes a second substrate 231 and a nanoarray structure disposed on the second substrate 231. The nanoarray structure includes multiple first nanopillars 232 and multiple nanoflow guides 233. The multiple first nanopillars 232 and multiple nanoflow guides 233 are arranged in multiple rows and alternately. The spacing between adjacent first nanopillars 232 and adjacent nanoflow guides 233 in each row gradually decreases along the virus suspension flow direction 234. The surface of the first nanopillars 232 is modified with target virus-specific capture antibodies. The fluorescence detection module 3 is disposed corresponding to the detection chamber 22 and is used to capture, process, and count fluorescence signals. The control system 4 is used to control the coordinated operation of the aerosol collection module 1, the microfluidic chip module 2, and the fluorescence detection module 3.

[0036] The aerosol collection module 1 can efficiently convert virus-laden aerosols in the air into a liquid virus suspension. The air inlet pipe 13 extends obliquely into the gas-liquid mixing chamber 11, ensuring that the high-speed airflow can obliquely impact the liquid surface to form a vortex and atomization mixing zone, thereby achieving efficient collection of aerosol viruses. The microfluidic chip module 2 can achieve precise delivery of the virus suspension and specific capture of virus particles. A gradually staggered nanoarray structure composed of multiple first nanopillars 232 and multiple nano-guide plates 233 is used. Specifically, the density of the first nanopillars 232 and nano-guide plates 233 gradually increases along the flow direction of the virus suspension, breaking the laminar flow state of the microfluidic liquid. The nano-guide plates 233 direct and divert the virus suspension, breaking up virus particle aggregates and preventing aggregation, thus achieving single-particle dispersion. Simultaneously, the collision of two intersecting liquid flows around the first nanopillars 232 creates turbulence, causing the virus particles in the suspension to rotate randomly, significantly increasing the contact probability between the virus particles and the surface of the first nanopillars 232, allowing the virus particles to be accurately captured on the first nanopillars 232. The fluorescence detection module 3, based on the specific binding principle of "first nanopillar 232 capture - virus - detection antibody - fluorescent group," accurately identifies the fluorescent spot corresponding to a single virus, achieving fluorescent labeling of the virus. Then, the fluorescence signal is collected, processed, and counted. The control system serves as the "central hub" of the entire testing device, enabling collaborative work and automated operation of all modules, thereby lowering the operational threshold.

[0037] The gas-liquid mixing chamber 11 is made of polytetrafluoroethylene (PTFE). PTFE is chemically inert and does not adsorb virus particles, thus preventing virus inactivation and loss. Preferably, the internal dimensions of the gas-liquid mixing chamber 11 are 2.5cm × 5cm, i.e., a diameter of 2.5cm and a height of 5cm, with an effective volume of 20mL. Preferably, the upper part of the gas-liquid mixing chamber 11 is cylindrical, and the lower part is hemispherical.

[0038] Preferably, the air inlet pipe 13 is made of quartz material, with an inner diameter of 2cm and an outer diameter of 3cm. The other end is connected to the aerosol sampling head 130 via a sealed connector. The inclination angle of the air inlet pipe 13 is 30°-45°, meaning the angle between the centerline of the air inlet pipe 13 and the vertical plane is 30°-45°. One end of the air inlet pipe 13 is 0.8cm-1.2cm from the inner wall of the gas-liquid mixing chamber 11, and the other end is 2.5cm-3.5cm from the liquid surface of the gas-liquid mixing chamber 11, further ensuring that the high-speed airflow can obliquely impact the liquid surface to form a vortex and atomized mixing zone. More preferably, the inclination angle of the air inlet pipe 13 is 40°, with one end 1cm from the inner wall of the gas-liquid mixing chamber 11 and the other end 3cm from the liquid surface of the gas-liquid mixing chamber 11. An air pump 131 and an airflow valve 132 are preferably installed on the air intake pipe 13. The air pump 131 pumps external aerosol into the gas-liquid mixing chamber 11 along the air intake pipe 13, and the airflow valve 132 is used to control the gas flow rate.

[0039] Preferably, the inlet tube 14 is made of medical-grade polyethylene (PE) tubing, with an inner diameter of 2 mm and an outer diameter of 3 mm. One end is connected to the storage bottle 12, and the other end is connected to the bottom of the gas-liquid mixing chamber 11. Preferably, the injected buffer solution is PBS buffer solution with a pH of 7.2-7.6, and the liquid volume is controlled to be less than 1 / 4 of the internal volume of the storage bottle 12. Preferably, a first peristaltic pump 141 is installed on the inlet tube 14.

[0040] Preferably, the top of the gas-liquid mixing chamber 11 is provided with an exhaust port 111 to maintain balance with atmospheric pressure.

[0041] A second peristaltic pump 151 is preferably provided on the guide tube 15, and the virus suspension is delivered to the detection chamber 22 of the microfluidic chip module 2 by the control of the second peristaltic pump 151.

[0042] The microfluidic chip module 2 also includes an inlet 24 and a delivery channel 25. The inlet 24 is connected to the other end of the guide tube 15. The two ends of the delivery channel 25 are connected to the inlet 24 and the detection chamber 22, respectively. A rinsing solution bottle 16 and a fluorescent labeling solution bottle 17 are also provided, both of which are connected to the guide tube 15. Preferably, the first substrate 21 is made of quartz material with a light transmittance ≥95% and dimensions of 20mm × 30mm × 5mm. The inlet 24, delivery channel 25, and detection chamber 22 are fabricated on the first substrate 21 using photolithography. Both the delivery channel 25 and the detection chamber 22 are hydrophobically treated to prevent liquid adhering to the walls and causing virus residue. A first branch pipe 161 connects the rinsing solution bottle 16 to the guide tube 15, and a third peristaltic pump 162 is installed on the first branch pipe 161. A second branch pipe 171 connects the fluorescent labeling solution bottle 17 to the guide tube 15, and a fourth peristaltic pump 172 is installed on the second branch pipe 171, for respectively allowing the rinsing solution and the fluorescent labeling solution to enter the detection chamber 22. The microfluidic chip module 2 also includes a drain port 26, which is connected to the detection chamber 22 and is connected to a collection bottle 27 for discharging and collecting waste liquid.

[0043] Preferably, the width of the delivery channel 25 is 80μm-200μm and the depth is 40μm-80μm. The dimensions of the detection chamber 22 are 4mm×4mm×0.4mm-6mm×6mm×0.6mm, and the effective volume of the detection chamber 22 is 8μL-20μL. More preferably, the width of the delivery channel 25 is 100μm and the depth is 50μm. Preferably, the dimensions of the detection chamber 22 are 5mm×5mm×0.5mm, and the effective volume is 12.5μL. Preferably, the biochip 23 is detachably installed in the detection chamber 22, and replacement does not require disassembly of the first substrate 21, facilitating the replacement of the biochip 23 with an operation time ≤30s. Specifically, the first substrate 21 is provided with a groove 211 communicating with the detection chamber 22, and the biochip 23 is inserted into the detection chamber 22 through the groove 211. To achieve a seal, the groove 211 is sealed by a flexible silicone cap 212. The preferred microfluidic chip module 2 also includes a cover plate 28, which covers the first substrate 21.

[0044] Please see Figure 3 Preferably, the height of the first nanopillar 232 and the nano-flow guide plate 233 are both 200nm-400nm, the diameter of the first nanopillar 232 is 80nm-120nm, the thickness of the nano-flow guide plate 233 is 80nm-120nm, and the spacing between adjacent first nanopillars 232 is 200nm-300nm. To facilitate the flow of the virus suspension, it is preferable that adjacent nano-flow guide plates 233 in each row form a figure-eight shape. A second nanopillar 235 is disposed between at least one adjacent nano-flow guide plate 233 in each row, which can better capture the virus.

[0045] The preferred second substrate 231 of the biochip 23 is a single-crystal silicon wafer with dimensions of 5mm × 5mm × 0.3mm. A nanoarray structure is fabricated on its surface using a photolithography-dry etching-wet cleaning process. The specific fabrication steps of the biochip 23 are as follows: ① A 1μm thick photoresist is coated onto the silicon wafer surface, followed by drying, exposure, and development to obtain a photoresist pattern; ② Inductively coupled plasma (ICP) dry etching is used to obtain the nanoarray structure; ③ BOE etching solution is used... =1:6, wet cleaning for 5s to remove residual oxide layer, finally ultrasonic cleaning with deionized water and drying with nitrogen. Preferably, the photoresist in step ① is AZ6130, the drying temperature is 90℃, the drying time is 60s, the ultraviolet light wavelength used for exposure is 365nm, the radiation intensity is 100mW / cm², and the developer is AZ300MIF, the developing temperature is 25℃, and the developing time is 60s. Preferably, the etching gas in step ② is... and The mixed gas has a volume ratio of 3:1, an etching power of 500W, and an etching time of 300s. Preferably, the cleaning power in step ③ is 50W, and the cleaning time is 30s. The nanoarray structure consists of several first nanopillars 232 and nano-guide plates 233 distributed in a gradually increasing manner along the flow direction of the virus suspension, with the density gradually increasing from front to back, and the final gap shrinking to 200nm. This ensures that each first nanopillar 232 can accommodate one or two virus particles, and the spacing between adjacent first nanopillars 232 is set to avoid virus aggregation. Virus capture methods include antibody capture, electrostatic capture, and chemical bonding. Precise binding is achieved by modifying the surfaces of the first nanopillar 232 and the second nanopillar 235 with target virus-specific capture antibodies via EDC / NHS activation. The specific steps are as follows: ① Immerse the biochip 23 in a 1% APTES (3-aminopropyltriethoxysilane) ethanol solution and incubate at room temperature to introduce a large number of amino groups, increasing the number of antibody binding sites; ② Immerse the biochip 23 in a pH 5.5 MES buffer, add 10 mM EDC and 5 mM NHS, and activate at room temperature for 30 min; ③ Add a first target virus-specific antibody, such as the novel coronavirus spike protein S1 antibody (catalog number: ab272504), and incubate at 4°C; ④ Rinse three times with PBS buffer for 5 min each time to remove unbound antibodies, and finally vacuum dry for storage. The biochip 23 is inserted into the detection chamber 22 via an embedded structure.

[0046] Fluorescent labeling solution: Detection antibodies modified with fluorescent groups can bind to viral surface antigen sites, causing viral labeling fluorescence. Preparation steps: ① Select a second target virus-specific detection antibody, such as an antibody against the nucleocapsid protein N of the novel coronavirus (catalog number: ab283773), and pretreat it by dialysis with carbonate buffer; ② Dissolve and activate fluorescein (FITC) in anhydrous DMSO, and couple it at a 1:8 molar ratio of antibody to fluorescein at 4°C in the dark with stirring for 3 hours, then terminate the reaction with aminoethanol; ③ Purify and remove free fluorescein using a Sephadex G-25 gel column, collect qualified conjugates (F / P value, fluorescein / protein molar ratio of 2-4), dilute to 0.1-0.5 mg / mL with PBS buffer containing 0.02% sodium azide at pH 7.4, and store in a sealed container in the dark.

[0047] The fluorescence detection module 3 includes a fluorescence excitation unit 31, an optical imaging unit 32, and a signal processing unit 33. The fluorescence excitation unit 31 includes a laser 311 and a focusing lens 312. The optical imaging unit 32 includes a beam splitter 321, a filter group, a fluorescence microscope 322, and a CCD camera 323. The signal processing unit 33 includes an image acquisition card 331 and a data processing module 332. The laser 311 serves as the excitation source, emitting excitation light 313. The wavelength of the excitation light matches the excitation wavelength of the fluorescein in the fluorescent labeling solution. For example, FITC is compatible with a 488nm excitation wavelength. The focusing lens 312 focuses the excitation light onto the nanoarray structure region of the detection chamber 22, ensuring concentrated excitation light energy and enhancing the fluorescence signal intensity. Preferably, the laser 311 is an ultraviolet-visible laser with an output power of 5-20mW (adjustable). The beam splitter 321 reflects the excitation light and transmits the target fluorescence. The filter group of the optical imaging unit 32 consists of an excitation filter 324 and a fluorescence filter 325. The excitation filter 324 only allows excitation light of a specific wavelength to pass through, while the fluorescence filter 325 filters background stray light and residual excitation light, retaining only the target fluorescence signal and reducing interference. The fluorescence microscope 322 has a magnification of 200×-400×, which is suitable for observing single virus fluorescence points and ensures that the fluorescence signal corresponding to a single virus can be clearly identified. The CCD camera 323 has a pixel count ≥10 million pixels, an exposure time of 10-50ms, a frame rate ≥15fps, and a quantum efficiency ≥80%. It has high resolution and high sensitivity, and can quickly capture fluorescence images of the nanoarray structure region, avoiding fluorescence signal attenuation. The image acquisition card 331 of the signal processing unit 3 converts the optical image captured by the CCD camera 323 into a digital signal. The data processing module 332 removes image noise through noise reduction algorithms such as median filtering, extracts the fluorescent dot region using a threshold segmentation algorithm, and eliminates false signals through morphological analysis. Finally, it achieves accurate counting of fluorescent dots, with each fluorescent dot corresponding to a captured virus particle.

[0048] Please see Figure 4 The control system 4 includes a core controller 41, a sensor group 42, a human-machine interface module 43, and a data storage module 44. The core controller 41 is a microcontroller or a programmable logic controller (PLC). The sensor group 42 includes a gas flow meter 421 and a liquid flow meter 422. The human-machine interface module 43 includes a touch screen and status indicator lights. The core controller 41 uses an industrial-grade microcontroller, such as the STM32 series, or a PLC, which has stable control performance and rich interfaces. It can simultaneously connect to and control the air pump 131, the first peristaltic pump 141, the second peristaltic pump 151, the third peristaltic pump 162, the fourth peristaltic pump 172, the fluorescence detection module 3, and other actuators, as well as the sensor group 42. The gas flow meter 421 monitors the airflow velocity in the inlet pipe 13 in real time, and the liquid flow meter 422 monitors the liquid delivery velocity. The data is fed back to the core controller 41 in real time. The core controller 41 automatically adjusts the actuators according to preset parameters to ensure that the parameters of each link are accurately met. Equipped with a 3.5-inch touchscreen display and status indicator lights, the touchscreen is used for parameter settings such as sampling time, delivery flow rate, incubation time, start / stop operation, and display of test results. The status indicator lights use different colors to distinguish the device's operating status, such as green - standby, yellow - sampling in progress, blue - testing in progress, red - malfunction, and white - completed. The data storage module 44 has a built-in storage chip that can store at least 1000 sets of test data, including test time, virus concentration, and fluorescence images. It supports data export via USB interface or wireless data transmission via Bluetooth or WiFi, facilitating data traceability and statistical analysis.

[0049] This application also discloses a method for detecting aerosol virus particles, using the aforementioned aerosol virus particle detection device, including the following steps:

[0050] (1) Aerosol collection: The buffer solution is injected into the gas-liquid mixing chamber 11 from the storage bottle 12. The control system 4 is started. Air is rushed into the gas-liquid mixing chamber 11 at an angle through the air inlet pipe 13. It forms a gas-liquid impact mixing with the injected buffer solution. The virus particles in the aerosol are captured into the buffer solution to form a virus suspension.

[0051] (2) Virus suspension delivery: The virus suspension is delivered from the gas-liquid mixing chamber 11 to the detection chamber 22 of the microfluidic chip module 2;

[0052] (3) Virus capture: The virus suspension comes into contact with the nanoarray structure of the biochip 23 in the detection chamber 22. The nanofluid guide plate 233 diverts the virus particles. The modified target virus-specific capture antibody on the surface of the first nanopillar 232 specifically binds to the first antigen on the surface of the virus particles to complete the virus capture.

[0053] (4) Fluorescent labeling: Fluorescent labeling solution is delivered to detection chamber 22. The fluorescent labeling detection antibody in the fluorescent labeling solution binds to the second antigen corresponding to the surface of the virus particles, and the captured virus is fluorescently labeled.

[0054] (5) Fluorescence detection: Fluorescence detection module 3 is activated to capture the fluorescence signal of the nanoarray structure, process the fluorescence signal and count the fluorescence spots;

[0055] (6) Result output: The control system 4 outputs the number or concentration of virus particles based on the fluorescence dot count results to complete the detection.

[0056] Preferably, after step (3) and before step (4), a rinsing process is also included: after virus capture is completed, rinsing fluid is introduced into the detection chamber 22 to rinse away unbound impurities, free viruses, and other interfering substances. This process improves the accuracy of subsequent virus detection.

[0057] To better illustrate the aerosol virus particle detection method of the present invention, the following are detailed steps.

[0058] 1. Device preparation: Select a biochip 23 that is compatible with the target virus, such as the novel coronavirus, and embed it into the detection chamber 22 of the microfluidic chip module 2, ensuring good sealing; inject PBS buffer solution of pH 7.4 into the storage bottle 12, inject PBS buffer solution of pH 7.4 into the rinsing bottle 16, and inject FITC-labeled detection antibody solution into the fluorescent labeling bottle 17; connect each tubing and check the sealing to avoid leakage.

[0059] 2. Parameter settings: Detection parameters can be set via the touch screen. The parameters are: aerosol collection time 8 min, air pump 131 airflow speed 50 L / min, virus suspension delivery flow rate 2 μL / min, virus capture incubation time 6 min, and fluorescent labeling incubation time 3 min. The parameters can be flexibly adjusted according to the target virus type and detection scenario.

[0060] 3. Aerosol collection: The control system 4 starts the air pump 131 and the first peristaltic pump 141. Air enters the gas-liquid mixing chamber 11 at a preset flow rate through the air inlet pipe 13, forming a strong gas-liquid impact and vortex mixing with the PBS buffer in 1 / 4 of its volume. The aerosol virus particles in the air are efficiently captured into the buffer to form a virus suspension. After collection is completed, the air pump 131 and the first peristaltic pump 141 are turned off.

[0061] 4. Virus suspension delivery: The second peristaltic pump 151 is started, and the virus suspension at the bottom of the gas-liquid mixing chamber 11 is slowly delivered to the detection chamber 22 of the microfluidic chip module 2 along the delivery channel 25. The virus suspension is delivered at a low speed, and the virus particles 291 are in full contact with the nanoarray structure.

[0062] 5. Virus capture: The virus suspension remains and incubates in the detection chamber 22. The multi-row first nanopillars 232 and multi-row nano-guide plates 233 of the nano-array structure divert and swirl the virus particles, preventing virus aggregation while ensuring full contact with the first nanopillars 232 and the second nanopillars 235 and interception. The modified specific antibodies on the surface of the first nanopillars 232 and the second nanopillars 235 specifically bind to the first antigen on the surface of the virus particles 291, achieving precise virus capture.

[0063] 6. Rinsing: After the virus capture is completed, the third peristaltic pump 162 corresponding to the rinsing solution is started to introduce PBS buffer into the detection chamber 22 at a flow rate of 10 μL / min for 2 min to rinse away unbound impurities, free viruses and other interfering substances. The waste liquid after rinsing is discharged through the drain port 26 of the microfluidic chip module 2 and collected in the collection bottle 27.

[0064] 7. Fluorescent labeling: The fourth peristaltic pump 172 corresponding to the fluorescent labeling solution is activated, and the fluorescent labeling detection antibody 292 solution is injected into the detection chamber 22 and incubated for 3 minutes. It specifically binds to the second antigen on the surface of the virus particles, so that each captured virus particle is fluorescently labeled 293. After incubation, the excess fluorescent labeling solution is drained and the rinsing step is repeated.

[0065] 8. Fluorescence Detection: When the fluorescence detection module 3 is activated, the excitation light 313 emitted by the laser 311 is focused onto the nanoarray structure region by the focusing lens 312, exciting the fluorescent marker to emit fluorescence; after the filter group filters out background stray light, the fluorescence microscope 322 amplifies the fluorescence signal, the CCD camera 323 captures the fluorescence image and transmits it to the signal processing unit 33, and the image acquisition card converts the optical image captured by the CCD camera 323 into a digital signal; the data processing module performs noise reduction, segmentation and fluorescence dot counting on the image.

[0066] 9. Result Output: The control system calculates the concentration or number of virus particles based on the fluorescence dot count and displays the test results on the touch screen. The entire test process takes 25 minutes. If the test result exceeds the preset threshold, the status indicator light will issue an alarm, and the data storage module 44 will automatically record the test data.

[0067] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

[0068] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. An aerosol virus particle detection device, characterized in that, include: An aerosol collection module includes a gas-liquid mixing chamber, a storage bottle, an air inlet pipe, a liquid inlet pipe, and a guide pipe. One end of the air inlet pipe extends into the gas-liquid mixing chamber and is inclined. Both ends of the liquid inlet pipe are connected to the gas-liquid mixing chamber and the storage bottle, respectively. One end of the guide pipe is connected to the bottom of the gas-liquid mixing chamber. A microfluidic chip module includes a first substrate, a detection chamber, and a biochip disposed within the detection chamber. The other end of a flow guide tube is connected to the detection chamber. The biochip includes a second substrate and a nanoarray structure disposed on the second substrate. The nanoarray structure includes multiple first nanopillars and multiple nanoflow guides. The multiple first nanopillars and multiple nanoflow guides are arranged in multiple rows and alternately. The spacing between adjacent first nanopillars and the spacing between adjacent nanoflow guides in each row gradually decreases along the direction of virus suspension flow. The surface of the first nanopillars is modified with a target virus-specific capture antibody. A fluorescence detection module, located corresponding to the detection chamber, is used to capture, process, and count fluorescence signals; The control system is used to control the coordinated operation of the aerosol collection module, the microfluidic chip module, and the fluorescence detection module.

2. The aerosol virus particle detection device according to claim 1, characterized in that, The inclination angle of the air inlet pipe is 30°-45°, one end of the air inlet pipe is 0.8cm-1.2cm away from the inner wall of the gas-liquid mixing chamber, and one end of the air inlet pipe is 2.5cm-3.5cm away from the liquid surface of the gas-liquid mixing chamber.

3. The aerosol virus particle detection device according to claim 1, characterized in that, The biochip can be detachably installed within the detection chamber.

4. The aerosol virus particle detection device according to claim 1, characterized in that, The microfluidic chip module also includes an inlet and a delivery channel. The inlet is connected to the other end of the guide tube. The two ends of the delivery channel are connected to the inlet and the detection chamber, respectively. It also includes a rinsing liquid bottle and a fluorescent labeling liquid bottle, both of which are connected to the guide tube.

5. The aerosol virus particle detection device according to claim 1, characterized in that, The height of the first nanopillar and the nanofluid plate is 200nm-400nm, the diameter of the first nanopillar is 80nm-120nm, the thickness of the nanofluid plate is 80nm-120nm, and the spacing between adjacent first nanopillars is 200nm-300nm.

6. An aerosol virus particle detection device according to claim 1 or 5, characterized in that, The adjacent nanofluids in each row form a figure-eight shape.

7. The aerosol virus particle detection device according to claim 6, characterized in that, A second nanopillar is disposed between at least one adjacent nanofluidic plate in each row.

8. The aerosol virus particle detection device according to claim 1, characterized in that, The fluorescence detection module includes a fluorescence excitation unit, an optical imaging unit, and a signal processing unit. The fluorescence excitation unit includes a laser and a focusing lens. The optical imaging unit includes a filter group, a fluorescence microscope, and a CCD camera. The signal processing unit includes an image acquisition card and a data processing module.

9. The aerosol virus particle detection device according to claim 1, characterized in that, The control system includes a core controller, a sensor group, a human-machine interaction module, and a data storage module. The core controller is a microcontroller or a programmable logic controller. The sensor group includes a gas flow meter and a liquid flow meter. The human-machine interaction module includes a touch screen and status indicator lights.

10. A method for detecting aerosol virus particles, characterized in that, Using an aerosol virus particle detection device as described in any one of claims 1-9 includes the following steps: (1) Aerosol collection: The buffer solution is injected into the gas-liquid mixing chamber from the storage bottle. The control system is started. Air is rushed into the gas-liquid mixing chamber at an angle through the air inlet pipe. It forms a gas-liquid impact mixing with the injected buffer solution. Virus particles in the aerosol are captured into the buffer solution to form a virus suspension. (2) Virus suspension delivery: The virus suspension is delivered from the gas-liquid mixing chamber to the detection chamber of the microfluidic chip module; (3) Virus capture: The virus suspension comes into contact with the nanoarray structure of the biochip in the detection chamber. The nanofluid plate diverts the virus particles. The modified target virus-specific capture antibody on the surface of the first nanopillar binds specifically to the first antigen on the surface of the virus particles to complete the virus capture. (4) Fluorescent labeling: Fluorescent labeling solution is delivered to the detection chamber. The fluorescent labeling detection antibody in the fluorescent labeling solution binds to the second antigen corresponding to the surface of the virus particles, and the captured virus is fluorescently labeled. (5) Fluorescence detection: The fluorescence detection module is activated to capture the fluorescence signal of the nanoarray structure, process the fluorescence signal and count the fluorescence spots; (6) Result output: The control system outputs the number or concentration of virus particles based on the fluorescence dot count results to complete the detection.

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