A microfluidic chip, a detection method and a device-driving-free allergen multi-checking microfluidic chip

Abstract

The application provides a microfluidic chip, a detection method and an allergen multi-connection detection microfluidic chip without equipment driving, and relates to the biomedical field.The microfluidic chip comprises a gas supply device and a chip main body, the chip main body is provided with a sample adding port, the inside of the chip main body is provided with an air channel, a plurality of sample inlet channels, a plurality of reaction flow channels and a reaction waste liquid containing cavity, the plurality of sample inlet channels are communicated with the sample adding port, the plurality of reaction flow channels are communicated with the plurality of sample inlet channels one by one, the plurality of reaction flow channels are communicated with the reaction waste liquid containing cavity, and the reaction flow channels are provided with different index allergen antigen spots.The gas supply device is arranged, and the gas supply device is communicated with the reaction flow channels through the air channel;after the reaction liquid reacts with the allergen antigen spots, the gas supply device inputs airflow into the reaction flow channels through the air channel, so that the reaction liquid is transported to the reaction waste liquid containing cavity, the residual fluorescent microspheres in the reaction flow channels are prevented from interfering with the detection of the allergen index, and the detection precision is improved.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, and more particularly to a microfluidic chip, a detection method, and a device-free microfluidic chip for multi-sensor detection of allergens. Background Technology

[0002] Allergies occur when the body's immune system malfunctions, causing an abnormal reaction to allergens. Currently, the main methods for allergen testing are divided into two categories: in vivo testing and in vitro testing. In vivo testing primarily uses skin scratch tests or patch tests to expose the allergen directly to the skin and observe for redness, swelling, rashes, or other symptoms to determine the allergy status. In vitro testing, on the other hand, involves detecting specific antibodies in the patient's blood or bodily fluids. Compared to in vivo testing, it is safer and suitable for most people.

[0003] In existing microfluidic chip detection devices, after the reaction solution completes, it remains in the self-driving channel and cannot be self-drained. The fluorescent microspheres in the reaction solution interfere with the detection of allergen indicators, affecting the detection accuracy. Summary of the Invention

[0004] This invention provides a microfluidic chip to solve the problem in existing detection devices where the reaction solution remains in the self-driving channel after the reaction is completed, and the fluorescent microspheres in the reaction solution interfere with the detection of allergen indicators.

[0005] This invention provides a microfluidic chip, comprising:

[0006] Gas supply device;

[0007] The chip body has a sample inlet and internally includes a gas channel, multiple sample inlet channels, multiple reaction channels, and a reaction waste liquid containment chamber. The multiple sample inlet channels are connected to the sample inlet, and the multiple reaction channels are connected to the reaction waste liquid containment chamber in a one-to-one correspondence. The reaction channels are equipped with different allergen antigen spots. A gas supply device is connected to the reaction channels through the gas channel. The gas supply device is used to supply airflow to the reaction channels through the gas channel after the reaction liquid reacts with the allergen antigen spots, thereby transporting the reaction liquid to the reaction waste liquid containment chamber.

[0008] According to a microfluidic chip provided by the present invention, the chip body includes a bottom sealing layer, a reaction layer, a sample injection layer and a gas channel layer stacked sequentially from bottom to top. The gas channel layer is provided with an air inlet. Both the gas channel layer and the sample injection layer are provided with a sample dispensing port and a plurality of quantitative cell air holes. The gas supply device is disposed on the side of the gas channel layer away from the sample injection layer. The gas supply device is connected to the gas channel through the air inlet. The plurality of quantitative cell air holes are connected to the plurality of sample injection channels one by one. The quantitative cell air holes are provided with a hydrophobic membrane.

[0009] According to a microfluidic chip provided by the present invention, the gas supply device includes an air bag, the air bag is fixedly connected to the side of the air channel layer opposite to the sample injection layer, the air bag is provided with an air outlet, and the air outlet of the air bag is connected to the air inlet.

[0010] According to a microfluidic chip provided by the present invention, the sample introduction channel includes:

[0011] Multiple quantitative cells are disposed on the side of the reaction layer facing the sample introduction layer;

[0012] Multiple resistance channels are disposed inside the reaction layer;

[0013] Multiple shut-off valves are disposed inside the reaction layer. One end of each shut-off valve is connected to a corresponding metering cell through multiple resistance channels, and the other end of each shut-off valve is connected to a corresponding reaction channel. The gas channel is connected to the other end of each shut-off valve.

[0014] According to a microfluidic chip provided by the present invention, the reaction layer is further provided with an injection cell, an injection channel and a sample waste liquid cell on the side facing the injection layer. The injection cell is connected to the quantitative cell through one of the injection channels and is connected to the sample dispensing port. The sample waste liquid cell is connected to the quantitative cell through another injection channel. Both the gas channel layer and the injection layer are provided with waste liquid cell pores, which are connected to the sample waste liquid cell. The waste liquid cell pores are provided with a hydrophobic membrane.

[0015] According to a microfluidic chip provided by the present invention, the flow resistance of the resistance channel is greater than the flow resistance of the sample injection channel.

[0016] According to a microfluidic chip provided by the present invention, the reaction waste liquid containing cavity includes:

[0017] A reaction waste liquid tank is located on the side of the reaction layer away from the sample introduction layer;

[0018] A transfer channel is provided on the side of the reaction layer away from the sample introduction layer, and one end of the transfer channel is connected to the reaction waste liquid tank;

[0019] A flow guide port penetrates the reaction layer and is connected to the other end of the transfer channel and the reaction flow channel, respectively.

[0020] According to a microfluidic chip provided by the present invention, the bottom sealing layer is provided with a plurality of pores, and the plurality of pores are connected to a plurality of reaction waste liquid pools in a one-to-one correspondence, and the pores are provided with a hydrophobic membrane.

[0021] The present invention also provides a device-free microfluidic chip for multi-sensor allergen detection, comprising a sampling drive tube and the microfluidic chip described in any of the above embodiments.

[0022] The present invention also provides a method for detecting a microfluidic chip, the method being based on the microfluidic chip described in any of the preceding claims, the method comprising:

[0023] Samples are quantitatively collected using a sampling tube assembly, which is then inserted into the port at the other end of the storage cylinder to puncture the second separator membrane, allowing the diluent to flow into the microsphere storage chamber.

[0024] Shake the sampling drive tube to mix the diluent, fluorescent microspheres, and sample to form a reaction solution;

[0025] Insert the outlet tube into the sample inlet and rotate the drive rod to input the reaction solution into the sample inlet, so that the reaction solution enters the quantitative cell and the sample waste cell respectively.

[0026] The reaction solution in the quantitative cell enters the corresponding reaction channel so that the human IgG antibody-fluorescent microsphere complex in the reaction solution reacts with the allergen antigen spot to form antigen-human IgG antibody-fluorescent microsphere complex.

[0027] The microfluidic chip is placed inside the instrument and the air bladder is squeezed. The air bladder inputs airflow into the reaction channel through the air passage to transport the remaining reaction liquid in the reaction channel to the corresponding reaction waste liquid pool.

[0028] The antigen-human IgG antibody-fluorescent microsphere complex within the reaction channel was detected using an instrument.

[0029] The microfluidic chip provided by this invention features a gas supply device that is connected to a reaction channel via an air passage. After the reaction liquid reacts with the allergen antigen spots, the gas supply device inputs airflow into the reaction channel through the air passage to transport the reaction liquid to the reaction waste liquid containment chamber. This avoids interference from residual fluorescent microspheres in the reaction channel on the detection of allergen indicators and improves detection accuracy. Attached Figure Description

[0030] To more clearly illustrate the technical solutions in this 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 some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0031] Figure 1 This is an exploded structural diagram of the sampling drive tube provided by the present invention.

[0032] Figure 2 This is a schematic diagram of the liquid storage cylinder provided by the present invention.

[0033] Figure 3 This is a schematic diagram of the structure of the device-free allergen multi-detection microfluidic chip provided by the present invention.

[0034] Figure 4 This is an exploded structural diagram of the microfluidic chip provided by the present invention.

[0035] Figure 5 This is a schematic diagram of the airway layer provided by the present invention.

[0036] Figure 6 This is a schematic diagram of the structure of the reaction layer provided by the present invention.

[0037] Figure 7 This is a partially enlarged structural diagram of the guide port provided by the present invention.

[0038] Figure 8 This is a schematic diagram of the inclined plane structure provided by the present invention.

[0039] Figure label:

[0040] 100. Sampling drive tube; 110. Drive rod; 111. Threaded push rod; 112. Piston; 113. Holding part; 120. Liquid storage cylinder; 121. First separator membrane; 122. Second separator membrane; 123. Third separator membrane; 124. Threaded cavity; 125. Microsphere storage chamber; 126. Diluent storage chamber; 130. Sampling tube assembly; 131. Sampling tube body; 132. Outlet tube; 133. Capillary tube; 134. Sealing ring;

[0041] 200. Microfluidic chip; 210. Gas supply device; 220. Sample inlet; 230. Gas channel; 240. Sample injection channel; 241. Quantitative cell; 242. Resistance channel; 243. Shut-off valve; 244. Sample injection cell; 245. Sample injection channel; 246. Sample waste liquid tank; 250. Reaction channel; 260. Reaction waste liquid container; 261. Reaction waste liquid tank; 262. Transfer channel; 263. Guide port; 264. Vent; 271. Bottom sealing layer; 272. Reaction layer; 273. Sample injection layer; 274. Gas channel layer; 275. Gas inlet; 276. Quantitative cell vent; 277. Shut-off valve vent; 278. Waste liquid tank vent;

[0042] 300. Sloping structure. Detailed Implementation

[0043] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0044] The following is combined with Figures 4-8 The specific structure and working principle of the microfluidic chip of the present invention are described.

[0045] like Figure 4 As shown, the microfluidic chip 200 includes a gas supply device 210 and a chip body. The chip body is provided with a sample inlet 220. The chip body is provided with a gas channel 230, multiple sample inlet channels 240, multiple reaction channels 250 and a reaction waste liquid containment chamber 260. The multiple sample inlet channels 240 are connected to the sample inlet 220. The multiple reaction channels 250 are connected to the multiple sample inlet channels 240 one by one. The multiple reaction channels 250 are connected to the reaction waste liquid containment chamber 260. The reaction channels 250 are provided with different allergen antigen spots. The gas supply device 210 is connected to the reaction channels 250 through the gas channel 230. The gas supply device 210 is used to input airflow into the reaction channels 250 through the gas channel 230 after the reaction liquid reacts with the allergen antigen spots, so as to transport the reaction liquid to the reaction waste liquid containment chamber 260.

[0046] The microfluidic chip 200 provided by the present invention includes a gas supply device 210, which is connected to a reaction channel 250 via an air passage 230. After the reaction liquid reacts with the allergen antigen spots, the gas supply device 210 inputs airflow into the reaction channel 250 through the air passage 230 to transport the reaction liquid to the reaction waste liquid receiving cavity 260. This avoids interference from the residual fluorescent microspheres in the reaction channel 250 on the detection of allergen indicators and improves the detection accuracy.

[0047] In one embodiment of the present invention, such as Figure 4 and Figure 5 As shown, the chip body includes a bottom sealing layer 271, a reaction layer 272, a sample injection layer 273, and a gas channel layer 274 stacked sequentially from bottom to top. The bottom sealing layer 271, reaction layer 272, sample injection layer 273, and gas channel layer 274 are all rectangular plates. However, the shape of these plates is not limited to this; other shapes can also be used. Adjacent plates are sealed together to prevent liquid or gas from flowing out between them. Adjacent plates are bonded together; alternatively, they can be integrally molded or connected using other methods.

[0048] The airway layer 274 is provided with an air inlet 275, which can be a blind hole or a through hole penetrating the airway layer 274. Both the airway layer 274 and the sample injection layer 273 are provided with a sample inlet 220 and multiple quantitative cell vents 276. Each of the airway layer 274 and the sample injection layer 273 has one sample inlet 220, which are coaxially arranged and connected with the sample inlets 220 of the airway layer 274 and the sample injection layer 273 to form a liquid channel. Each of the airway layer 274 and the sample injection layer 273 has three quantitative cell vents 276. Of course, the number of quantitative cell vents 276 is not limited to this, and can also be four, five or more. The three quantitative cell vents 276 are arranged at intervals along the width direction of the airway layer 274 and the sample injection layer 273. The distance between two adjacent quantitative cell vents 276 can be the same or different, depending on the position of the quantitative cell 241. The quantitative cell vent 276 on the airway layer 274 is coaxially arranged and connected with the quantitative cell vent 276 on the sample injection layer 273 to form the exhaust channel of the quantitative cell 241. The quantitative cell vent 276 is provided with a hydrophobic membrane.

[0049] The sample injection layer 273 is provided with three shut-off valve ports 277, which are through holes. The three shut-off valve ports 277 are arranged at intervals along the width direction of the sample injection layer 273. Of course, the number of shut-off valve ports 277 is not limited to this; there can be four, five, or more, depending on the number of reaction channels 250. The gas channel layer 274 is provided with three gas channels 230 on the side facing the sample injection layer 273. One end of the three gas channels 230 is connected to the gas inlet 275, and the other end of the three gas channels 230 is connected to the three shut-off valve ports 277 one by one.

[0050] The gas supply device 210 is located on the side of the gas channel layer 274 away from the sample injection layer 273, and the gas supply device 210 is connected to the gas channel 230 through the gas inlet 275. Multiple quantitative cell gas holes 276 are connected to multiple sample injection channels 240 in a one-to-one correspondence. Specifically, the three quantitative cell gas holes 276 are connected to the three quantitative cells 241 in a one-to-one correspondence.

[0051] In one embodiment of the present invention, such as Figure 4 As shown, the gas supply device 210 includes an airbag, which is fixedly connected to the side of the airway layer 274 opposite to the sample injection layer 273. The airbag has an air outlet that communicates with the air inlet 275. Specifically, the airbag has an air outlet on the side facing the airway layer 274, and a limiting groove is provided on the side of the airway layer 274 opposite to the sample injection layer 273. The airbag is embedded in the limiting groove and fixed in the limiting groove by adhesive. The air outlet of the airbag is aligned with and communicates with the air inlet 275. By setting the limiting groove, the airbag can be limited, preventing displacement during use and improving the stability of the airbag.

[0052] Furthermore, the airbag is elliptical in shape, with a long side of 25 mm, a short side of 15 mm, a height of 8 mm, and contains approximately 2 mL of compressible air.

[0053] In one embodiment of the present invention, such as Figure 6 As shown, the sample injection channel 240 includes multiple quantitative cells 241, multiple resistance channels 242, and multiple shut-off valves 243. The multiple quantitative cells 241 are disposed on the side of the reaction layer 272 facing the sample injection layer 273, and are spaced apart along the width of the reaction layer 272. In this embodiment, three quantitative cells 241 are provided; however, the number of quantitative cells 241 is not limited to this and is determined based on the number of reaction channels 250. The three quantitative cells 241 are interconnected so that the reaction solution can flow between the multiple quantitative cells 241, ensuring that the volume of the reaction solution in each quantitative cell 241 is the same.

[0054] Multiple resistance channels 242 are disposed inside the reaction layer 272, and the multiple resistance channels 242 are arranged at intervals along the width direction of the reaction layer 272. In this embodiment, three resistance channels 242 are provided. Of course, the number of resistance channels 242 is not limited to this, and is determined according to the number of metering cells 241. The three resistance channels 242 are arranged one-to-one with the three metering cells 241. The cross-sectional area of ​​the resistance channels 242 is small to increase the flow resistance of the reaction liquid within the resistance channels 242. In this embodiment, the resistance channels 242 are arranged horizontally. Of course, the resistance channels 242 can also be inclined at a certain angle to further increase the flow resistance of the reaction liquid.

[0055] Multiple shut-off valves 243 are disposed inside the reaction layer 272, spaced apart along the width direction and along the thickness direction (vertical direction) of the reaction layer 272. In this embodiment, three shut-off valves 243 are provided; however, the number is not limited to this and is determined based on the number of metering cells 241. One end of each shut-off valve 243 is connected to one of the metering cells 241 via multiple resistance channels 242, and the other end is connected to one of the reaction channels 250. Gas passages 230 are connected to the other ends of the shut-off valves 243; specifically, three shut-off valve ports 277 are connected to the other ends of the three shut-off valves 243. By setting a shut-off valve 243 between the resistance flow channel 242 and the reaction flow channel 250, it is possible to further prevent the reaction liquid in the metering tank 241 from entering the reaction flow channel 250 under capillary action before the metering is completed. That is, after the metering of the reaction liquid is completed, the reaction liquid will break through the shut-off valve 243 under capillary action.

[0056] The microfluidic chip 200 provided by this invention, through the combined design of a metering cell 241, a shut-off valve 243, and a reaction channel 250, achieves multi-channel reaction, improving the accuracy of the liquid volume participating in the reaction in each reaction channel and avoiding dispensing errors. Under the pressure drive of the front-end sampling drive tube 100, the reaction liquid is pushed into the first metering cell 241. Gas in the first metering cell 241 is discharged from the metering cell vent 276, filling the first metering cell 241 with reaction liquid and achieving quantitative measurement of the reaction liquid. Since the flow resistance of the resistance channel 242 is greater than that of the injection channel 245, the remaining reaction liquid continues to enter the second and third metering cells 241 under pressure drive, completing the quantitative measurement based on the same principle as the first metering cell 241.

[0057] In one embodiment of the present invention, such as Figure 6 As shown, the reaction layer 272 facing the sample injection layer 273 also includes an injection tank 244, an injection channel 245, and a sample waste liquid tank 246. The injection tank 244 is connected to the quantitative tank 241 through an injection channel 245, which is a narrow trough. The position of the injection tank 244 corresponds to the position of the sample inlet 220. The injection tank 244 is connected to the sample inlet 220. After the reaction solution enters through the sample inlet 220, it first enters the injection tank 244 and then enters the quantitative tank 241 through the injection channel 245.

[0058] The sample waste liquid reservoir 246 is used to contain the excess reaction liquid after the quantitative reservoir 241 has completed quantitative measurement. The sample waste liquid reservoir 246 is connected to the third quantitative reservoir 241 through another sample inlet channel 245. Both the gas channel layer 274 and the sample inlet layer 273 are provided with waste liquid reservoir vents 278. The positions of the waste liquid reservoir vents 278 correspond to the positions of the sample waste liquid reservoir 246 and are connected to the sample waste liquid reservoir 246. The waste liquid reservoir vents 278 are provided with a hydrophobic membrane to facilitate the discharge of air from the waste liquid reservoir vents 278, so that the sample waste liquid reservoir 246 can contain more reaction liquid.

[0059] In one embodiment of the present invention, the cross-sectional area of ​​the resistance channel 242 is smaller than that of the sample inlet channel 245, and the flow resistance of the resistance channel 242 is greater than that of the sample inlet channel 245. By making the flow resistance of the resistance channel 242 greater than that of the sample inlet channel 245, the reaction solution cannot pass through the resistance channel 242 before the three quantitative cells 241 have completed quantification. Instead, it can only enter the other quantitative cells 241 to fill them. Finally, any excess reaction solution enters the sample waste liquid pool 246 through another sample inlet channel 245.

[0060] In one embodiment of the present invention, the reaction channel 250 is formed by connecting multiple S-shaped channels in sequence, and the reaction channel 250 with multiple bends is used to increase the length of the reaction channel 250.

[0061] The microfluidic chip 200 provided by this invention enables independent driving of multiple reaction channels 250, reducing the possibility of mutual interference between indicators. When the number of indicators is large, the detection of different allergens is easily affected by interference from adjacent indicators. To avoid this, the microfluidic chip 200 provided by this invention sets up three parallel and independent reaction channels 250, reducing the number of indicators arranged in a single reaction channel 250 while maintaining the same number of detectable indicators on a single chip. Combined with fluid self-driving, it achieves independent driving of the fluid within a single reaction channel 250, unaffected by interference from other channels.

[0062] In one embodiment of the present invention, such as Figure 6 As shown, the reaction waste liquid containing cavity 260 includes a reaction waste liquid tank 261, a transfer channel 262, and a guide port 263. The reaction waste liquid tank 261 is disposed on the side of the reaction layer 272 opposite to the sample injection layer 273. Multiple reaction waste liquid tanks 261 are provided, spaced apart along the width direction of the reaction layer 272. In this embodiment, three reaction waste liquid tanks 261 are provided. Of course, the number of reaction waste liquid tanks 261 is not limited to this; four, five, or more can also be provided, depending on the number of reaction channels 250. The reaction waste liquid tank 261 is used to contain the reaction liquid after the reaction, therefore, the volume of the reaction waste liquid tank 261 needs to be as large as possible.

[0063] A transfer channel 262 is located on the side of the reaction layer 272 opposite to the sample introduction layer 273, with one end connected to the reaction waste liquid tank 261. The transfer channel 262 guides the reaction solution after the reaction into the reaction waste liquid tank 261. The width of the transfer channel 262 is small to utilize capillary action to guide the reaction solution into the reaction waste liquid tank 261. In this embodiment, three transfer channels 262 are provided. Of course, the number of transfer channels 262 is not limited to this; four, five, or more can be provided, depending on the number of reaction channels 250. A guide port 263 connects the other end of the transfer channel 262 to the reaction channel 250. The guide port 263 penetrates the reaction layer 272 and is connected to both the other end of the transfer channel 262 and the reaction channel 250.

[0064] In a preferred embodiment of the present invention, since the reaction waste liquid tank 261 and the reaction flow channel 250 are located on both sides of the reaction layer 272, a black opaque material is used to separate the reaction waste liquid tank 261 and the reaction flow channel 250. On the one hand, the interference of excitation scattered light during reaction signal detection can be avoided, and on the other hand, the fluorescence of the signal substance in the waste liquid in the closed environment is prevented from being excited by the excitation light that has been projected, thus avoiding interference with the detection signal.

[0065] In a preferred embodiment of the present invention, such as Figure 7 As shown, the longitudinal cross-section of the guide port 263 is frustum-shaped, meaning the inner diameter of the guide port 263 near the reaction channel 250 is larger than the inner diameter near the transfer channel 262. This design allows the reaction liquid to be driven from top to bottom under hydrophilic action. The reaction liquid spontaneously moves towards the area where the channel contracts, ensuring a continuous flow of the reaction liquid to the side of the reaction layer 272 opposite to the sample injection layer 273.

[0066] In a preferred embodiment of the present invention, the bottom sealing layer 271 is provided with a plurality of vents 264, which are connected one-to-one with a plurality of reaction waste liquid pools 261. Each vent 264 is provided with a hydrophobic membrane. By providing a hydrophobic membrane in the vents 264, gas in the reaction waste liquid pools 261 can be discharged, allowing more reaction liquid to enter the reaction waste liquid pools 261.

[0067] The microfluidic chip 200 provided by this invention has lower reagent costs compared to other detection methods such as microfluidic chemiluminescence, and a single chip can detect up to 72 indicators. There are currently no similar multi-detection products in the existing technology, and the cost is only one-tenth to one-hundredth of other detection devices.

[0068] The microfluidic chip 200 provided by this invention has extremely low requirements for the instrument's optical path, and signal reading can be performed using image methods. Users can also directly interpret the results by irradiating with an ultraviolet lamp, eliminating the need for detection equipment. The liquid inside the chip does not require instrument driving, further reducing the cost of supporting instruments. Compared to other detection devices, the cost of the microfluidic chip 200 provided by this invention is only one-thousandth of theirs.

[0069] like Figure 3 As shown, the present invention also provides a device-free allergen multi-detection microfluidic chip 200, which includes a sampling drive tube 100 and the microfluidic chip 200 described in any of the above embodiments.

[0070] The device-free, multi-sensor allergen detection microfluidic chip 200 provided by this invention integrates multiple steps required for analysis, such as quantitative sample collection, sample dilution, fluorescent microsphere reconstitution, reagent mixing, sample addition, and liquid-driven operation, through simple fluid manipulation combined with a self-driving principle. This completes the immune response for multiple pre-embedded allergen indicators within the chip. It simplifies the often cumbersome process of a dozen or even twenty steps to four, reducing the demands on testing personnel and significantly shortening the waiting time for test reports. For multi-sensor testing—specific IgG antibody detection—the reaction time is reduced from 2-4 hours to approximately 20 minutes, improving detection efficiency.

[0071] The following is combined with Figures 1-2 The specific structure and working principle of the sampling drive transistor of the present invention are described.

[0072] like Figure 1 and Figure 2 As shown, the sampling drive tube 100 includes a drive rod 110, a liquid storage cylinder 120, and a sampling tube assembly 130. The liquid storage cylinder 120 is internally provided with a first separating membrane 121, a second separating membrane 122, and a third separating membrane 123 arranged sequentially along the length of the liquid storage cylinder 120, dividing the interior of the liquid storage cylinder 120 into a threaded cavity 124, a microsphere storage chamber 125, and a diluent storage chamber 126. The microsphere storage chamber 125 stores fluorescent microspheres. Compared to the traditional design of placing dried fluorescent microspheres in the chip reaction area, this invention stores the dried fluorescent microspheres in the microsphere storage chamber 125, shifting the reaction of the fluorescent microspheres forward, ensuring sufficient reaction time between the fluorescent microspheres and human IgG in the sample. Shaking and mixing further ensures the full binding of the fluorescent microspheres to the human IgG in the sample. The diluent storage chamber 126 stores diluent. The drive rod 110 is inserted into the threaded cavity 124 through the port at one end of the liquid storage cylinder 120 and is threaded into the threaded cavity 124. The sampling tube assembly 130 is inserted into the port at the other end of the liquid storage cylinder 120.

[0073] The sampling drive tube 100 provided by this invention, due to the threaded engagement of the drive rod 110 with the threaded cavity 124, increases the length of the drive rod 110 entering the liquid storage cylinder 120 by rotating the drive rod 110 during liquid injection. This allows for precise control of the injection of the reaction liquid into the microfluidic chip 200 through the sampling tube assembly 130, avoiding excessively fast flow rates and improving detection accuracy. Compared to external air pumps for propulsion, the sampling drive tube 100 of this invention eliminates the need for a power unit, reducing costs and eliminating issues such as contamination and aerosol pollution.

[0074] In one embodiment of the present invention, the lengths of the threaded cavity 124, the microsphere storage chamber 125, and the diluent storage chamber 126 are all different, with the threaded cavity 124 having the longest length. The specific length of the threaded cavity 124 is determined based on the length of the drive rod 110.

[0075] In one embodiment of the present invention, the drive rod 110 includes a threaded push rod 111 and a piston 112. The outer peripheral surface of the threaded push rod 111 is provided with an external thread, and the inner wall of the threaded cavity 124 is provided with an internal thread. The outer diameter of the threaded push rod 111 is larger than the outer diameter of the piston 112. The internal thread is located on the inner wall of the threaded cavity 124 at the end away from the microsphere storage chamber 125. The threaded push rod 111 is threadedly engaged with the inner wall of the threaded cavity 124.

[0076] The piston 112 has a rod-like structure and is coaxially arranged with the threaded push rod 111. The piston 112 and the threaded push rod 111 are connected at the ends near the sampling tube assembly 130. Preferably, the piston 112 and the threaded push rod 111 are integrally formed. The piston 112 slides and seals against the inner wall of the threaded cavity 124. Specifically, an annular protrusion is provided at the end of the piston 112 away from the threaded push rod 111 to increase the outer diameter of the end of the piston 112 away from the threaded push rod 111. The annular protrusion slides against the inner wall of the threaded cavity 124, and at the same time, the annular protrusion seals against the inner wall of the threaded cavity 124. By rotating the drive rod 110, the length of the drive rod 110 entering the threaded cavity 124 can be changed, thereby increasing the pressure inside the liquid storage cylinder 120. Under the action of pressure, the reaction liquid flows out through the sampling tube assembly 130.

[0077] In one embodiment of the present invention, the drive rod 110 further includes a gripping portion 113 located outside the liquid storage cylinder 120. The gripping portion 113 is cylindrical, but its shape is not limited to this; it can also be ellipsoidal or other shapes. The outer diameter of the gripping portion 113 is larger than the outer diameter of the threaded push rod 111 to increase the contact area of ​​the gripping portion 113, making it easier for the operator to grip. The gripping portion 113 is connected to the end of the threaded push rod 111 away from the sampling tube assembly 130. Specifically, the gripping portion 113 and the threaded push rod 111 are integrally formed. Of course, the connection method between the gripping portion 113 and the threaded push rod 111 is not limited to this; a threaded connection or other methods can also be used for connection.

[0078] In one embodiment of the present invention, the sampling tube assembly 130 includes a sampling tube body 131 and an outlet tube 132. The sampling tube body 131 is a hollow tube, and its outer diameter is slightly smaller than the inner diameter of the liquid storage cylinder 120. The sampling tube body 131 is hollow inside and is inserted into the port at the other end of the liquid storage cylinder 120 to seal the other end of the liquid storage cylinder 120. The outlet tube 132 is inserted into the sample inlet to output the reaction solution. The outlet tube 132 is a tube with a smaller outer diameter and is coaxially arranged with the sampling tube body 131. The outlet tube 132 is connected to the end of the sampling tube body 131 away from the drive rod 110. The outlet tube 132 and the sampling tube body 131 are integrally formed or bonded together.

[0079] In one embodiment of the present invention, the sampling tube assembly 130 further includes a capillary 133, which is connected to one end of the sampling tube body 131 near the drive rod 110. The capillary 133 is coaxially arranged with the sampling tube body 131, and has a small inner diameter to allow the sample to enter the capillary 133 under the action of the capillary. The capillary 133 is used to collect samples and puncture the second septum 122 to connect the microsphere storage chamber 125 with the diluent storage chamber 126. The length of the capillary 133 determines the amount of sample that can be collected, and the length of the capillary 133 is specifically set according to experimental requirements. By setting the capillary 133 inside the sampling tube body 131, the accuracy of the amount of liquid involved in the reaction is ensured, while the requirements on the usage environment and users are reduced. Users do not need to repeatedly add samples with a pipette, making it more convenient for home use.

[0080] In one embodiment of the present invention, a bevel is provided at one end of the capillary 133 near the drive rod 110. The bevel is provided so that the second separator membrane 122 can be pierced and the capillary can draw the sample into the capillary 133.

[0081] In one embodiment of the present invention, the sampling tube assembly 130 further includes a plurality of connecting pieces, which are spaced apart inside the sampling tube body 131 and surround the outer periphery of the capillary 133. The distance between two adjacent connecting pieces is equal, and the connecting pieces are connected to the inner wall of the capillary 133 and the sampling tube body 131. Preferably, the connecting pieces are integrally formed with the capillary 133 and the inner wall of the sampling tube body 131. A flow channel is formed between two adjacent connecting pieces to communicate the outlet tube 132 with the diluent storage chamber 126.

[0082] The sampling drive tube 100 of this invention simplifies and integrates the structure of the blood collection and sample loading device, realizing integrated operation of the blood collection and sample loading device. Compared with the traditional sample pretreatment process, it greatly reduces the user's operating difficulty and the error caused by multiple sample loadings. The traditional sample pretreatment process requires centrifugation to remove serum after blood collection, then quantitatively adding the sample to the centrifuge tube, adding diluent and mixing, and then using a pipette to add it to the reagent card. In contrast, the sampling drive tube 100 of this invention directly inserts into the diluent storage chamber 126 after drawing whole blood, shakes, inserts the entire device into the microfluidic chip 200, and rotates the drive rod. The entire process requires no precise operation and has a high fault tolerance.

[0083] In one embodiment of the present invention, an annular groove is provided on the outer peripheral surface of the sampling tube body 131, and a sealing ring 134 is provided in the annular groove. The sealing ring 134 is made of rubber or silicone. The sampling tube body 131 is sealed to the port of the other end of the liquid storage cylinder 120 through the sealing ring 134.

[0084] In one embodiment of the present invention, the first separator 121 is a hydrophobic membrane that separates the threaded cavity 124 from the microsphere storage chamber 125. The first separator 121 allows air to pass through but prevents liquid from passing through. The second separator 122 and the third separator 123 are both metal membranes, preferably aluminum membranes.

[0085] The present invention also provides a detection method for a microfluidic chip 200, the detection method being based on the microfluidic chip 200 described above, and the detection method comprising:

[0086] In step S100, a sample is quantitatively collected through the sampling tube assembly 130, and the sampling tube assembly 130 is inserted into the port at the other end of the liquid storage cylinder 120 to puncture the second separator membrane 122, so that the diluent flows into the microsphere storage chamber.

[0087] Specifically, firstly, quantitative sample collection is performed through capillary 133. The third separator membrane 123 is torn open, and the sampling tube assembly 130 is inserted into the port at the other end of the liquid storage cylinder 120. One end of the capillary 133 is inserted into the diluent storage chamber 126, and the sampling tube body 131 is closed so that the capillary 133 punctures the second separator membrane 122, releasing the diluent (the amount of diluent used is 100-1000 μL, and the type is phosphate solution) that has been pre-stored in the diluent storage chamber 126. After the diluent is released, it flows into the microsphere storage chamber.

[0088] Step S200: Shake the sampling drive tube 100 to mix the diluent, fluorescent microspheres and sample to form a reaction solution;

[0089] Specifically, the sampling drive tube 100 is inverted and shaken to mix the diluent, fluorescent microspheres, and sample in the microsphere storage chamber to form a reaction solution. During the mixing process, the human IgG antibody in the sample combines with the fluorescent microspheres labeled with anti-human IgG secondary antibody to form a human IgG antibody-fluorescent microsphere complex, which is uniformly distributed in the reaction solution along with the remaining unreacted sample and microspheres.

[0090] In step S300, the outlet tube 132 is inserted into the sample inlet 220 and the drive rod 110 is rotated to input the reaction solution into the sample inlet 220, so that the reaction solution enters the quantitative cell 241 and the sample waste cell 246 respectively.

[0091] Specifically, the outlet tube 132 is inserted into the sample inlet 220, and the drive rod 110 is rotated to drive the piston 112 to slowly move towards the sampling tube assembly 130, adding the reaction solution into the sample inlet 244. The reaction solution flows along the sample inlet channel 240 under the drive of the piston to the three quantitative cells 241 until the three quantitative cells 241 are filled. The excess reaction solution enters the sample waste liquid pool 246 through another sample inlet channel 240 until the hydrophobic membrane in the pore 278 of the waste liquid pool is blocked, thus completing the addition and separation of the reaction solution.

[0092] In step S400, the reaction solution in the quantitative cell 241 enters the corresponding reaction channel 250 so that the human IgG antibody-fluorescent microsphere complex in the reaction solution reacts with the allergen antigen spot to form antigen-human IgG antibody-fluorescent microsphere complex.

[0093] Specifically, under capillary action, the quantitative reaction solution in the three quantitative pools 241 passes through the stop valve 243 along the resistance flow channel 242 and enters the reaction flow channel 250. The human IgG antibody-fluorescent microsphere complex in the reaction solution binds to the allergen antigen spots pre-embedded in the reaction flow channel 250, forming an antigen-human IgG antibody-fluorescent microsphere complex immobilized on the chip. The remaining sample and fluorescent microspheres that did not participate in the reaction continue to flow with the reaction solution. When the reaction solution reaches the guide port 263, due to the hydrophilic effect of the guide port 263, the reaction solution fills the guide port 263 and is transferred to the lower transfer channel 262, and under capillary action, enters the reaction waste pool 261 along the transfer channel 262, completing the immune reaction process.

[0094] In step S500, the microfluidic chip 200 is placed into the instrument and the air bag is squeezed. The air bag inputs airflow into the reaction channel 250 through the air channel 230 to transport the remaining reaction liquid in the reaction channel 250 to the corresponding reaction waste liquid pool 261.

[0095] Specifically, after the capillary self-driven reaction is completed, the microfluidic chip 200 is inserted into the matching detection instrument. The inclined structure 300 at the instrument inlet compresses the air bladder, causing the gas in the bladder to enter the reaction channel 250 along the air passage 230. This drives the residual reaction liquid in the reaction channel 250 to the reaction waste liquid pool 261 at the end of the transfer channel 262, completing the waste discharge step. For example... Figure 8 As shown, the inclined surface of the inclined structure 300 faces downwards. As the airbag enters the gap below the inclined surface to a greater depth, the gap becomes smaller, causing the airbag to be gradually compressed.

[0096] In step S600, the antigen-human IgG antibody-fluorescent microsphere complex in the reaction channel 250 is detected using an instrument.

[0097] Specifically, the instrument uses a semiconductor laser to emit a laser of a specific wavelength to irradiate the reaction channel 250. The antigen-human IgG antibody-fluorescent microsphere complex within the reaction channel 250 emits a fluorescent signal. The instrument converts the fluorescent signal emitted by the detection chip into a photoelectric signal through the image sensor in the camera module, forming a two-dimensional image signal. The image information is then converted and the signal value is output to achieve the detection of different allergen indicators.

[0098] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A microfluidic chip, characterized in that, include: Gas supply device (210); The chip body is provided with a sample inlet (220). The chip body is provided with an air channel (230), multiple sample inlet channels (240), multiple reaction channels (250), and a reaction waste liquid containment chamber (260). The multiple sample inlet channels (240) are connected to the sample inlet (220). The multiple reaction channels (250) are connected to the multiple sample inlet channels (240) one by one. The multiple reaction channels (250) are connected to the reaction waste liquid containment chamber (260). The reaction channels (250) are provided with different allergen antigen spots. The gas supply device (210) is connected to the reaction channels (250) through the air channel (230). The gas supply device (210) is used to input airflow into the reaction channels (250) through the air channel (230) after the reaction liquid reacts with the allergen antigen spots, so as to transport the reaction liquid to the reaction waste liquid containment chamber (260). The chip body includes a bottom sealing layer (271), a reaction layer (272), a sample injection layer (273), and a gas channel layer (274) stacked from bottom to top, and the gas channel layer (274) is provided with an air inlet (275). The gas supply device (210) includes an air bag, which is fixedly connected to the side of the airway layer (274) away from the sample injection layer (273). The air bag is provided with an air outlet, which is connected to the air inlet (275). The sample inlet channel (240) includes: Multiple quantitative cells (241) are disposed on the side of the reaction layer (272) facing the injection layer (273); Multiple resistance channels (242) are disposed inside the reaction layer (272); Multiple shut-off valves (243) are disposed inside the reaction layer (272). One end of each shut-off valve (243) is connected to a plurality of metering cells (241) through multiple resistance channels (242). The other end of each shut-off valve (243) is connected to a plurality of reaction channels (250). The gas channel (230) is connected to the other end of each shut-off valve (243). The reaction layer (272) is further provided with an injection pool (244), an injection channel (245), and a sample waste liquid pool (246) on the side facing the injection layer (273). The injection pool (244) is connected to the quantitative pool (241) through one of the injection channels (245), and the injection pool (244) is connected to the sample dispensing port (220). The sample waste liquid pool (246) is connected to the quantitative pool (241) through another injection channel (245). The flow resistance of the resistance channel (242) is greater than that of the injection channel (245).

2. The microfluidic chip according to claim 1, characterized in that, Both the airway layer (274) and the sample injection layer (273) are provided with the sample inlet (220) and multiple quantitative cell pores (276). The multiple quantitative cell pores (276) are connected to the multiple sample injection channels (240) one by one, and the quantitative cell pores (276) are provided with hydrophobic membranes.

3. The microfluidic chip according to claim 2, characterized in that, Both the airway layer (274) and the sample injection layer (273) are provided with waste liquid pool vents (278), the waste liquid pool vents (278) are connected to the sample waste liquid pool (246), and the waste liquid pool vents (278) are provided with hydrophobic membranes.

4. The microfluidic chip according to claim 2 or 3, characterized in that, The reaction waste liquid containment chamber (260) includes: A reaction waste liquid tank (261) is disposed on the side of the reaction layer (272) away from the sample injection layer (273); A transfer channel (262) is provided on the side of the reaction layer (272) away from the sample introduction layer (273), and one end of the transfer channel (262) is connected to the reaction waste liquid tank (261); The flow guide (263) penetrates the reaction layer (272) and is connected to the other end of the transfer channel (262) and the reaction channel (250).

5. The microfluidic chip according to claim 4, characterized in that, The bottom sealing layer (271) is provided with a plurality of pores (264), and the plurality of pores (264) are connected to the plurality of reaction waste liquid pools (261) in a one-to-one correspondence. The pores (264) are provided with a hydrophobic membrane.

6. A device-free microfluidic chip for multi-sensor allergen detection, characterized in that, It includes a sampling drive transistor (100) and a microfluidic chip as described in any one of claims 1 to 5.

7. A method for detecting a microfluidic chip, said detection method being based on the microfluidic chip according to any one of claims 1 to 5, characterized in that, The detection method includes: Samples are quantitatively collected by sampling tube assembly (130), and the sampling tube assembly (130) is inserted into the port at the other end of the liquid storage cylinder (120) to puncture the second separator membrane (122) so that the diluent flows into the microsphere storage chamber; Shake the sampling drive tube (100) to mix the diluent, fluorescent microspheres and sample to form a reaction solution; Insert the outlet tube (132) into the sample inlet (220) and rotate the drive rod (110) to input the reaction solution into the sample inlet (220), so that the reaction solution enters the quantitative cell (241) and the sample waste cell (246) respectively. The reaction solution in the quantitative cell (241) enters the corresponding reaction channel (250) so that the human IgG antibody-fluorescent microsphere complex in the reaction solution reacts with the allergen antigen spot to form antigen-human IgG antibody-fluorescent microsphere complex; The microfluidic chip is placed inside the instrument and the air bag is squeezed. The air bag inputs airflow into the reaction channel (250) through the air channel (230) to transport the remaining reaction liquid in the reaction channel (250) to the corresponding reaction waste liquid pool (261). The antigen-human IgG antibody-fluorescent microsphere complex in the reaction channel (250) was detected using an instrument.

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