Micro-fluidic chip, detection method and equipment-driving-free allergen multi-joint detection micro-fluidic chip

Abstract

The invention provides a micro-fluidic chip, a detection method and an allergen multi-joint detection micro-fluidic chip free of equipment driving, and relates to the field of biomedicine. The micro-fluidic chip comprises a gas supply device and a chip main body, the chip main body is provided with a sample adding port, the chip main body is internally provided with a gas channel, a plurality of sample introduction channels, a plurality of reaction runners and a reaction waste liquid accommodating cavity, the plurality of sample introduction channels are communicated with the sample adding port, the plurality of reaction runners are communicated with the plurality of sample introduction channels in a one-to-one correspondence manner, and the reaction waste liquid accommodating cavity is communicated with the gas channel. The plurality of reaction runners are communicated with the reaction waste liquid accommodating cavity, and the reaction runners are provided with allergen antigen spots with different indexes. The gas supply device is arranged, and the gas supply device is communicated with the reaction flow channel through the gas channel; after the reaction liquid reacts with allergen antigen spots, the gas supply device inputs gas flow into the reaction flow channel through the gas channel, so that the reaction liquid is conveyed to the reaction waste liquid containing cavity, interference of residual fluorescent microspheres in the reaction flow channel on detection of allergen indexes is avoided, and the detection precision is improved.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a microfluidic chip, a detection method and an allergen multi-detection microfluidic chip without equipment drive. Background Art

[0002] Allergy is caused by abnormal reaction to allergens due to the malfunction of human immune system. At present, the main allergen detection methods are divided into two categories: in vivo detection and in vitro detection. Among them, in vivo detection is mainly carried out through skin scratch test or patch test, which makes allergens directly contact with human body and observes whether redness, swelling, rash and other phenomena appear on the surface of skin to judge the allergy. In vitro detection is carried out by specific antibody detection of patients' blood, body fluids, etc. Compared with in vivo detection, it is safer and suitable for most people.

[0003] After the reaction of the reaction liquid in the existing microfluidic chip detection device is completed, the reaction liquid will remain in the self-driving channel and cannot be self-emptied. The fluorescent microspheres in the reaction liquid interfere with the detection of allergen indicators and affect the detection accuracy. Summary of the invention

[0004] The present invention provides a microfluidic chip for solving the problem that after the reaction of the reaction liquid in the existing detection device is completed, the reaction liquid will stay in the self-driving channel and the fluorescent microspheres in the reaction liquid will interfere with the detection of allergen indicators.

[0005] The present invention provides a microfluidic chip, comprising: Gas supply device; The chip body is provided with a sample addition port, and an airway, a plurality of injection channels, a plurality of reaction channels and a reaction waste liquid holding chamber are provided inside the chip body. The plurality of injection channels are connected with the sample addition port, the plurality of reaction channels are connected with the plurality of injection channels in a one-to-one correspondence, the plurality of reaction channels are connected with the reaction waste liquid holding chamber, and the reaction channels are provided with allergen antigen spots of different indicators; the air supply device is connected with the reaction channel through the airway, and the air supply device is used to input airflow to the reaction channel through the airway after the reaction liquid reacts with the allergen antigen spots, so as to transport the reaction liquid to the reaction waste liquid holding chamber.

[0006] According to a microfluidic chip provided by the present invention, the chip body includes a bottom sealing layer, a reaction layer, an injection layer and an airway layer stacked in sequence from bottom to top, the airway layer is provided with an air inlet hole, the airway layer and the injection layer are both provided with the sample addition port and a plurality of quantitative pool air holes, the air supply device is arranged on a side of the airway layer away from the injection layer, the air supply device is connected with the airway through the air inlet hole, a plurality of the quantitative pool air holes are connected with a plurality of the injection channels in a one-to-one correspondence, and the quantitative pool air holes are provided with a hydrophobic membrane.

[0007] According to a microfluidic chip provided by the present invention, the air supply device includes an air bag, the air bag is fixedly connected to the side of the airway layer away from the sample introduction layer, the air bag is provided with an air outlet, and the air outlet of the air bag is connected to the air inlet.

[0008] According to a microfluidic chip provided by the present invention, the injection channel comprises: A plurality of quantitative pools are arranged on a side of the reaction layer facing the injection layer; A plurality of resistance flow channels are arranged inside the reaction layer; A plurality of stop valves are arranged inside the reaction layer, one end of the plurality of stop valves is connected to the plurality of quantitative pools one-to-one through the plurality of resistance flow channels, the other end of the plurality of stop valves is connected to the plurality of reaction flow channels one-to-one, and the gas channel is connected to the other end of the plurality of stop valves.

[0009] According to a microfluidic chip provided by the present invention, a sampling pool, a sampling flow channel and a sample waste liquid pool are also provided on the side of the reaction layer facing the sampling layer, the sampling pool is connected to the quantitative pool through one of the sampling flow channels, the sampling pool is connected to the sample addition port, the sample waste liquid pool is connected to the quantitative pool through another of the sampling flow channels, the airway layer and the sampling layer are both provided with waste liquid pool pores, the waste liquid pool pores are connected to the sample waste liquid pool, and the waste liquid pool pores are provided with a hydrophobic membrane.

[0010] According to a microfluidic chip provided by the present invention, the flow resistance of the resistance flow channel is greater than the flow resistance of the injection flow channel.

[0011] According to a microfluidic chip provided by the present invention, the reaction waste liquid receiving chamber comprises: A reaction waste liquid pool is arranged on a side of the reaction layer away from the injection layer; A transfer channel is arranged on a side of the reaction layer away from the injection layer, and one end of the transfer channel is connected to the reaction waste liquid pool; A flow guide port, wherein the flow guide port penetrates the reaction layer and is respectively connected to the other end of the transfer channel and the reaction flow channel.

[0012] According to a microfluidic chip provided by the present invention, the bottom sealing layer is provided with a plurality of pores, the plurality of pores are connected with the plurality of reaction waste liquid pools in a one-to-one correspondence, and the pores are provided with a hydrophobic membrane.

[0013] The present invention also provides a device-driven-free allergen multi-detection microfluidic chip, comprising a sampling drive tube and any of the above-mentioned microfluidic chips.

[0014] The present invention also provides a method for detecting a microfluidic chip, the method being based on the microfluidic chip described in any one of the above, the method comprising: Collecting the sample quantitatively through the sampling tube assembly, and inserting the sampling tube assembly into the port at the other end of the liquid storage cylinder to puncture the second separation membrane, so that the diluent flows into the microsphere storage chamber; Shake the sampling drive tube to mix the diluent, fluorescent microspheres and sample to form a reaction solution; Insert the outlet tube into the sample injection port and rotate the driving rod to input the reaction liquid into the sample injection port, so that the reaction liquid enters the quantitative pool and the sample waste liquid pool respectively; The reaction liquid in the quantitative pool enters the corresponding reaction channel, so that the human IgG antibody-fluorescent microsphere complex in the reaction liquid reacts with the allergen antigen spot to form an antigen-human IgG antibody-fluorescent microsphere complex; The microfluidic chip is placed in the instrument and the air bag is squeezed, and the air bag inputs airflow to the reaction channel through the air channel to transport the remaining reaction liquid in the reaction channel to the corresponding reaction waste liquid pool; The antigen-human IgG antibody-fluorescent microsphere complex in the reaction flow channel is detected by an instrument.

[0015] The microfluidic chip provided by the present invention is provided with an air supply device, and the air supply device is connected with the reaction channel through an airway; after the reaction liquid reacts with the allergen antigen spot, the air supply device inputs airflow to the reaction channel through the airway to transport the reaction liquid to the reaction waste liquid holding chamber, thereby avoiding interference of residual fluorescent microspheres in the reaction channel with the detection of allergen indicators and improving the detection accuracy. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0017] Figure 1 It is a schematic diagram of the exploded structure of the sampling drive tube provided by the present invention.

[0018] Figure 2 It is a structural schematic diagram of the liquid storage cylinder provided by the present invention.

[0019] Figure 3 It is a schematic diagram of the structure of the device-driven-free allergen multi-detection microfluidic chip provided by the present invention.

[0020] Figure 4 It is a schematic diagram of the exploded structure of the microfluidic chip provided by the present invention.

[0021] Figure 5 It is a schematic diagram of the structure of the airway layer provided by the present invention.

[0022] Figure 6 It is a schematic structural diagram of the reaction layer provided by the present invention.

[0023] Figure 7 It is a schematic diagram of the partially enlarged structure of the guide port provided by the present invention.

[0024] Figure 8 It is a structural schematic diagram of the inclined surface structure provided by the present invention.

[0025] Reference numerals: 100, sampling drive tube; 110, drive rod; 111, threaded push rod; 112, piston; 113, gripping part; 120, liquid storage cylinder; 121, first partition membrane; 122, second partition membrane; 123, third partition 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; 200, microfluidic chip; 210, gas supply device; 220, sample addition port; 230, airway; 240, sample injection channel; 241, quantitative pool; 242, resistance flow channel; 243, stop valve; 244, sample injection pool; 245, sample injection flow channel; 246, sample waste liquid pool; 250, reaction flow channel; 260, reaction waste liquid holding chamber; 261, reaction waste liquid pool; 262, transfer channel; 263, flow guide port; 264, air hole; 271, bottom sealing layer; 272, reaction layer; 273, sample injection layer; 274, airway layer; 275, air inlet; 276, quantitative pool air hole; 277, stop valve air port; 278, waste liquid pool air hole; 300. Sloped structure. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] Combine the following Figure 4-Figure 8 The specific structure and working principle of the microfluidic chip of the present invention are described.

[0028] like Figure 4 As shown, the microfluidic chip 200 includes an air supply device 210 and a chip body. The chip body is provided with a sample addition port 220. The chip body is provided with an air channel 230, a plurality of injection channels 240, a plurality of reaction channels 250 and a reaction waste liquid holding chamber 260. The plurality of injection channels 240 are connected with the sample addition port 220. The plurality of reaction channels 250 are connected with the plurality of injection channels 240 in a one-to-one correspondence. The plurality of reaction channels 250 are connected with the reaction waste liquid holding chamber 260. The reaction channels 250 are provided with allergen antigen spots of different indicators. The air supply device 210 is connected with the reaction channel 250 through the air channel 230. The air supply device 210 is used to input airflow to the reaction channel 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 holding chamber 260.

[0029] The microfluidic chip 200 provided by the present invention is provided with a gas supply device 210, and the gas supply device 210 is connected with the reaction channel 250 through the air channel 230; after the reaction liquid reacts with the allergen antigen spot, the gas supply device 210 inputs airflow to the reaction channel 250 through the air channel 230 to transport the reaction liquid to the reaction waste liquid holding chamber 260, thereby avoiding the interference of the fluorescent microspheres remaining in the reaction channel 250 on the detection of the allergen index, and improving the detection accuracy.

[0030] In one embodiment of the present invention, Figure 4 and Figure 5 As shown, the chip body includes a bottom sealing layer 271, a reaction layer 272, a sampling layer 273 and an airway layer 274 stacked in sequence from bottom to top, and the bottom sealing layer 271, the reaction layer 272, the sampling layer 273 and the airway layer 274 are all rectangular plates. Of course, the shape of the above-mentioned plate is not limited thereto, and other shapes can also be used. The two adjacent plates are sealed to prevent liquid or gas from flowing out through the two adjacent plates. The two adjacent plates are bonded and connected, and of course, they can also be connected by one-piece molding or other connection methods.

[0031] Airway layer 274 is provided with air inlet 275, and air inlet 275 can be a blind hole, and can also be a through hole running through airway layer 274. Airway layer 274 and sampling layer 273 are all provided with sample adding port 220 and multiple quantitative pool air holes 276, and airway layer 274 and sampling layer 273 are respectively provided with a sample adding port 220, and the sample adding port 220 of airway layer 274 is coaxially arranged with the sample adding port 220 of sampling layer 273 and is communicated to form liquid passage. Airway layer 274 and sampling layer 273 are respectively provided with three quantitative pool air holes 276, and certainly, the quantity of quantitative pool air holes 276 is not limited thereto, and can also be four, five or more quantities. Three quantitative pool air holes 276 are arranged at intervals along the width direction of airway layer 274 and sampling layer 273, and the distance between adjacent two quantitative pool air holes 276 can be the same, and can also be different, and is specifically determined according to the position of quantitative pool 241. The quantitative pool pores 276 on the air channel layer 274 are coaxially arranged and connected with the quantitative pool pores 276 on the sample introduction layer 273 to form an exhaust channel of the quantitative pool 241. The quantitative pool pores 276 are provided with a hydrophobic membrane.

[0032] The sampling layer 273 is provided with three stop valve gas ports 277, which are through holes. The three stop valve gas ports 277 are arranged at intervals along the width direction of the sampling layer 273. Of course, the number of the stop valve gas ports 277 is not limited thereto, and may also be four, five or more, which is specifically determined according to the number of reaction channels 250. The gas channel layer 274 is provided with three gas channels 230 on one side facing the sampling 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 stop valve gas ports 277 in a one-to-one correspondence.

[0033] The gas supply device 210 is disposed on the side of the air channel layer 274 away from the sampling layer 273, and the gas supply device 210 is connected to the air channel 230 through the air inlet 275. The multiple quantitative pool air holes 276 are connected to the multiple sampling channels 240 in a one-to-one correspondence, and specifically, the three quantitative pool air holes 276 are connected to the three quantitative pools 241 in a one-to-one correspondence.

[0034] In one embodiment of the present invention, Figure 4 As shown, the air supply device 210 includes an airbag, which is fixedly connected to the side of the airway layer 274 away from the sample feeding layer 273, and the airbag is provided with an air outlet, and the air outlet of the airbag is connected with the air inlet 275. Specifically, the airbag is provided with 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 away from the sample feeding layer 273. The airbag is embedded in the limiting groove and fixed in the limiting groove by gluing. The air outlet of the airbag is aligned with and connected with the air inlet 275. By providing the limiting groove, the airbag can be limited, which can prevent the airbag from deflecting during use and improve the stability of the airbag.

[0035] Furthermore, the airbag is oval in shape, the long side of the airbag is 25 mm, the short side of the airbag is 15 mm, the height of the airbag is 8 mm, and the airbag contains about 2 mL of compressible air.

[0036] In one embodiment of the present invention, Figure 6 As shown, the injection channel 240 includes a plurality of quantitative pools 241, a plurality of resistance flow channels 242 and a plurality of stop valves 243. The plurality of quantitative pools 241 are arranged on the side of the reaction layer 272 facing the injection layer 273. The plurality of quantitative pools 241 are arranged at intervals along the width direction of the reaction layer 272. In this embodiment, three quantitative pools 241 are arranged; of course, the number of quantitative pools 241 is not limited thereto, and is specifically determined according to the number of reaction flow channels 250. The three quantitative pools 241 are interconnected so that the reaction liquid can flow between the plurality of quantitative pools 241, so that the volume of the reaction liquid in each quantitative pool 241 is the same.

[0037] A plurality of resistance flow channels 242 are arranged inside the reaction layer 272, and the plurality of resistance flow channels 242 are arranged at intervals along the width direction of the reaction layer 272. In this embodiment, three resistance flow channels 242 are arranged. Of course, the number of resistance flow channels 242 is not limited thereto, and is specifically determined according to the number of quantitative pools 241. The three resistance flow channels 242 are arranged one-to-one corresponding to the three quantitative pools 241, and the cross-sectional area of ​​the resistance flow channel 242 is small to increase the flow resistance of the reaction liquid in the resistance flow channel 242. In this embodiment, the resistance flow channel 242 is arranged horizontally. Of course, the resistance flow channel 242 can also be inclined at a certain angle to further increase the flow resistance of the reaction liquid.

[0038] A plurality of stop valves 243 are arranged inside the reaction layer 272, and the plurality of stop valves 243 are arranged at intervals along the width direction of the reaction layer 272, and the stop valves 243 are arranged along the thickness direction of the reaction layer 272, that is, arranged along the up and down direction. In the present embodiment, three stop valves 243 are provided, and of course, the number of stop valves 243 is not limited thereto, and is specifically determined according to the number of quantitative pools 241. One end of the plurality of stop valves 243 is connected to the plurality of quantitative pools 241 in a one-to-one correspondence through the plurality of resistance flow channels 242, and the other end of the plurality of stop valves 243 is connected to the plurality of reaction flow channels 250 in a one-to-one correspondence. The air channel 230 is connected to the other end of the plurality of stop valves 243, and specifically, the three stop valve air ports 277 are connected to the other end of the three stop valves 243 in a one-to-one correspondence. By setting a stop valve 243 between the resistance flow channel 242 and the reaction flow channel 250, the reaction liquid in the quantitative pool 241 can be further prevented from entering the reaction flow channel 250 under capillary action before the quantification is completed, that is, after the quantification of the reaction liquid is completed, the reaction liquid breaks through the stop valve 243 under capillary action.

[0039] The microfluidic chip 200 provided by the present invention realizes multi-channel reaction through the combined design of quantitative pool 241, stop valve 243 and reaction channel 250, improves the accuracy of the liquid capacity involved in the reaction in each reaction channel, and avoids the occurrence of liquid separation error. The reaction liquid is pushed into the first quantitative pool 241 under the pressure drive of the front sampling drive tube 100, and the gas in the first quantitative pool 241 is discharged from the quantitative pool pore 276, so that the reaction liquid is filled with the first quantitative pool 241, and the quantitative reaction liquid is realized. Since the flow resistance of the resistance flow channel 242 is greater than the flow resistance of the injection flow channel 245, the remaining reaction liquid continues to enter the second quantitative pool 241 and the third quantitative pool 241 under pressure drive, and the quantitative reaction is completed according to the same principle as the first quantitative pool 241.

[0040] In one embodiment of the present invention, Figure 6 As shown, a sample injection pool 244, a sample injection channel 245 and a sample waste liquid pool 246 are also provided on one side of the reaction layer 272 facing the sample injection layer 273. The sample injection pool 244 is connected to the quantitative pool 241 through a sample injection channel 245, and the sample injection channel 245 is a slot body with a relatively small width. The position of the sample injection pool 244 corresponds to the position of the sample injection port 220, and the sample injection pool 244 is connected to the sample injection port 220. After the reaction liquid enters through the sample injection port 220, it first enters the sample injection pool 244, and then enters the quantitative pool 241 through the sample injection channel 245.

[0041] The sample waste liquid pool 246 is used to accommodate excess reaction liquid after the quantitative pool 241 completes the quantitative measurement. The sample waste liquid pool 246 is connected to the third quantitative pool 241 through another injection channel 245. The air channel layer 274 and the injection layer 273 are both provided with waste liquid pool air holes 278. The position of the waste liquid pool air holes 278 corresponds to the position of the sample waste liquid pool 246 and is connected to the sample waste liquid pool 246. The waste liquid pool air holes 278 are provided with hydrophobic membranes to facilitate the discharge of air in the waste liquid pool air holes 278 through the waste liquid pool air holes 278, so that the sample waste liquid pool 246 can accommodate more reaction liquid.

[0042] In one embodiment of the present invention, the cross-sectional area of ​​the resistance flow channel 242 is smaller than the cross-sectional area of ​​the injection flow channel 245, and the flow resistance of the resistance flow channel 242 is greater than the flow resistance of the injection flow channel 245. By making the flow resistance of the resistance flow channel 242 greater than the flow resistance of the injection flow channel 245, before the three quantitative pools 241 complete the quantitative measurement, the reaction liquid cannot pass through the resistance flow channel 242, and can only enter other quantitative pools 241 to fill the quantitative pools 241, and finally the excess reaction liquid enters the sample waste liquid pool 246 through another injection flow channel 245.

[0043] In one embodiment of the present invention, the reaction channel 250 is formed by connecting a plurality of S-shaped channels in sequence. The reaction channel 250 having a plurality of bends is used to increase the length of the reaction channel 250 .

[0044] The microfluidic chip 200 provided by the present invention realizes independent driving of multiple reaction channels 250, reducing the possibility of mutual interference between indicators. When the number of indicators is too large, the detection of different allergens is susceptible to interference from adjacent indicators. To avoid this situation, the microfluidic chip 200 provided by the present invention is provided with three independent reaction channels 250 arranged side by side, thereby reducing the number of indicators arranged in a single reaction channel 250 while ensuring that the number of detectable indicators on a single chip remains unchanged, and combined with fluid self-driving, the independent driving of the fluid in a single reaction channel 250 is realized without interference from other channels.

[0045] In one embodiment of the present invention, Figure 6 As shown, the reaction waste liquid holding chamber 260 includes a reaction waste liquid pool 261, a transfer channel 262 and a guide port 263. The reaction waste liquid pool 261 is arranged on the side of the reaction layer 272 away from the sample injection layer 273. There are multiple reaction waste liquid pools 261, and the multiple reaction waste liquid pools 261 are arranged at intervals along the width direction of the reaction layer 272. In this embodiment, there are three reaction waste liquid pools 261. Of course, the number of reaction waste liquid pools 261 is not limited to this, and four, five or more numbers can also be set, which is specifically determined according to the number of reaction channels 250. The reaction waste liquid pool 261 is used to hold the reaction liquid after the reaction, so the volume of the reaction waste liquid pool 261 needs to be as large as possible.

[0046] The transfer channel 262 is arranged on the side of the reaction layer 272 away from the injection layer 273, and one end of the transfer channel 262 is connected to the reaction waste liquid pool 261. The transfer channel 262 is used to guide the reaction liquid after the reaction to the reaction waste liquid pool 261. The width of the transfer channel 262 is small, so as to guide the reaction liquid after the reaction to the reaction waste liquid pool 261 by capillary action. In this embodiment, three transfer channels 262 are provided. Of course, the number of transfer channels 262 is not limited thereto, and four, five or more can also be provided, which is specifically determined according to the number of reaction channels 250. The guide port 263 is used to connect the other end of the transfer channel 262 to the reaction channel 250. The guide port 263 penetrates the reaction layer 272, and the guide port 263 is respectively connected to the other end of the transfer channel 262 and the reaction channel 250.

[0047] In a preferred embodiment of the present invention, since the reaction waste liquid pool 261 and the reaction channel 250 are respectively located on both sides of the reaction layer 272, by using a black opaque material interval between the reaction waste liquid pool 261 and the reaction 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 signal substance in the waste liquid in a closed environment can be prevented from being excited to fluoresce by the projected excitation light, thereby interfering with the detection signal.

[0048] In a preferred embodiment of the present invention, Figure 7 As shown, the longitudinal cross section of the flow guide 263 is truncated cone-shaped, that is, the inner diameter of the flow guide 263 near the reaction channel 250 is larger than the inner diameter near the transfer channel 262. Such a design can drive the reaction liquid from top to bottom under the hydrophilic effect. The reaction liquid moves spontaneously to the place where the flow channel contracts, ensuring that the reaction liquid continuously flows to the side of the reaction layer 272 away from the injection layer 273.

[0049] In a preferred embodiment of the present invention, 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, and a hydrophobic membrane is provided in the pores 264. By providing the hydrophobic membrane in the pores 264, the gas in the reaction waste liquid pool 261 can be discharged, so that more reaction liquid can enter the reaction waste liquid pool 261.

[0050] The microfluidic chip 200 provided by the present 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 is currently no similar multi-detection product in the prior art, and the cost is only one-tenth to one-hundredth of that of other detection devices.

[0051] The microfluidic chip 200 provided by the present invention has extremely low requirements for the instrument optical path, and can adopt the image method for signal reading. Users can also directly use ultraviolet light to judge the results without the need for detection equipment. The liquid in the chip does not need to be driven by the instrument, further reducing the cost of the supporting instrument. Compared with other detection equipment, the cost of the microfluidic chip 200 provided by the present invention is only one thousandth of that.

[0052] like Figure 3 As shown, the present invention further provides a device-free allergen multiplex detection microfluidic chip 200, which comprises a sampling drive tube 100 and the microfluidic chip 200 described in any one of the above embodiments.

[0053] The device-free allergen multi-test microfluidic chip 200 provided by the present invention integrates multiple steps such as quantitative sample collection, sample dilution, fluorescent microsphere re-dissolution, reaction reagent mixing, sample addition, and liquid drive required for analysis through simple fluid manipulation combined with the principle of self-driving, and completes the immune response of multiple pre-embedded allergen indicators in the chip. Simplifying a dozen or even twenty operation steps to four steps reduces the requirements for detection operators and greatly shortens the waiting time for test reports. For the multi-test project - specific IgG antibody detection, the reaction time is shortened from 2-4 hours to about 20 minutes, and the detection efficiency is improved.

[0054] Combine the following Figure 1-Figure 2The specific structure and working principle of the sampling drive tube of the present invention are described.

[0055] 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 interior of the liquid storage cylinder 120 is provided with a first partition membrane 121, a second partition membrane 122 and a third partition membrane 123 arranged in sequence along the length direction of the liquid storage cylinder 120, so as to divide the interior of the liquid storage cylinder 120 into a threaded cavity 124, a microsphere storage chamber 125 and a diluent storage chamber 126 in sequence. Fluorescent microspheres are stored in the microsphere storage chamber 125. Compared with the traditional design of placing the dried fluorescent microspheres in the chip reaction area, the present invention advances the reaction of the fluorescent microspheres by storing the dried fluorescent microspheres in the microsphere storage chamber 125, thereby ensuring the reaction time of the fluorescent microspheres and the human IgG in the sample, and further ensuring the full combination of the fluorescent microspheres and the human IgG in the sample by shaking and mixing. The diluent storage chamber 126 stores diluent. The driving rod 110 is inserted into the threaded cavity 124 through a port at one end of the liquid storage cylinder 120 and threadably matches with the threaded cavity 124 . The sampling tube assembly 130 is inserted into a port at the other end of the liquid storage cylinder 120 .

[0056] The sampling drive tube 100 provided by the present invention has a threaded fit between the drive rod 110 and the threaded cavity 124. When injecting liquid, the drive rod 110 is rotated, and the length of the drive rod 110 entering the liquid storage cylinder 120 increases under the action of the thread, thereby accurately controlling the reaction liquid to be injected into the microfluidic chip 200 through the sampling tube assembly 130, thereby avoiding the reaction liquid from flowing too fast and improving the detection accuracy. Compared with the external air pump of the instrument for propulsion, the sampling drive tube 100 of the present invention does not need to be provided with a power device, which reduces the cost and does not have the problems of carryover pollution and aerosol pollution.

[0057] In one embodiment of the present invention, the length of the threaded cavity 124 , the length of the microsphere storage chamber 125 , and the length of the diluent storage chamber 126 are different, among which the length of the threaded cavity 124 is the largest. The specific length of the threaded cavity 124 is determined according to the length of the driving rod 110 .

[0058] In one embodiment of the present invention, the driving 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, wherein the outer diameter of the threaded push rod 111 is greater than the outer diameter of the piston 112, the internal thread is located on the inner wall of the threaded cavity 124 away from the microsphere storage chamber 125, and the threaded push rod 111 is threadedly matched with the inner wall of the threaded cavity 124.

[0059] The piston 112 is a rod-shaped structure. The piston 112 is coaxially arranged with the threaded push rod 111. The piston 112 is connected to one end of the threaded push rod 111 close to the sampling tube assembly 130. Preferably, the piston 112 and the threaded push rod 111 are integrally formed. The piston 112 slides and seals with the inner wall of the threaded cavity 124. Specifically, an annular convex ridge 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 ridge slides with the inner wall of the threaded cavity 124. At the same time, the annular ridge seals with the inner wall of the threaded cavity 124. By rotating the driving rod 110, the length of the driving rod 110 entering the threaded cavity 124 can be changed, so that the pressure inside the liquid storage cylinder 120 increases. Under the action of the pressure, the reaction liquid flows out through the sampling tube assembly 130.

[0060] In one embodiment of the present invention, the driving rod 110 further includes a holding portion 113, the holding portion 113 is located outside the liquid storage cylinder 120, and the holding portion 113 is cylindrical. Of course, the shape of the holding portion 113 is not limited thereto, and may also be an ellipsoid or other shapes. The outer diameter of the holding portion 113 is greater than the outer diameter of the threaded push rod 111 to increase the contact area of ​​the holding portion 113, so that the operator can hold it conveniently. The holding portion 113 is connected to one end of the threaded push rod 111 away from the sampling tube assembly 130. Specifically, the holding portion 113 is integrally formed with the threaded push rod 111. Of course, the connection method of the holding portion 113 and the threaded push rod 111 is not limited thereto, and may also be connected by threaded connection or other methods.

[0061] 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 body, and the outer diameter of the sampling tube body 131 is slightly smaller than the inner diameter of the liquid storage cylinder 120. The interior of the sampling tube body 131 is hollow, and the sampling tube body 131 is inserted into the port at the other end of the liquid storage cylinder 120 to block the port at the other end of the liquid storage cylinder 120. The outlet tube 132 is inserted into the sample addition port to output the reaction liquid. The outlet tube 132 is a tube body with a smaller outer diameter. The outlet tube 132 is coaxially arranged with the sampling tube body 131, and the outlet tube 132 is connected to the end of the sampling tube body 131 away from the driving rod 110. The outlet tube 132 and the sampling tube body 131 are integrally formed or bonded.

[0062] In one embodiment of the present invention, the sampling tube assembly 130 also includes a capillary 133, which is connected to one end of the sampling tube body 131 close to the driving rod 110. The capillary 133 is coaxially arranged with the sampling tube body 131, and the inner diameter of the capillary 133 is small so that the sampled sample can enter the capillary 133 under the capillary action. The capillary 133 is used to collect samples and puncture the second separation membrane 122 so that the microsphere storage chamber 125 is connected to the diluent storage chamber 126. The length of the capillary 133 determines the amount of collected samples that can be accommodated, and the length of the capillary 133 is specifically set according to the experimental requirements. By arranging the capillary 133 in the sampling tube body 131, the accuracy of the amount of liquid involved in the reaction is guaranteed while reducing the requirements for the use environment and users. Users do not need to repeatedly add samples with a pipette, which is more convenient for users to use at home.

[0063] In one embodiment of the present invention, a slope is provided at one end of the capillary 133 close to the driving rod 110 , and the slope is provided to facilitate piercing the second separation membrane 122 and to facilitate the capillary to absorb the sample into the capillary 133 .

[0064] In one embodiment of the present invention, the sampling tube assembly 130 further includes a plurality of connecting pieces, which are arranged at intervals inside the sampling tube body 131, and are arranged around the outer circumference of the capillary tube 133, with the distance between two adjacent connecting pieces being equal, and the connecting pieces are connected to the inner wall of the capillary tube 133 and the sampling tube body 131; preferably, the connecting pieces are integrally formed with the inner wall of the capillary tube 133 and the sampling tube body 131. A flow channel is formed between two adjacent connecting pieces to connect the outlet tube 132 with the diluent storage chamber 126.

[0065] The sampling drive tube 100 of the present invention simplifies and merges the structure of the blood sampling and sampling device, and realizes the integrated operation of the blood sampling and sampling device. Compared with the sample pretreatment process of traditional reagents, the user's operating difficulty and the error caused by multiple sampling are greatly reduced. In the traditional sample pretreatment process, after blood collection, it is necessary to first centrifuge to remove the serum, then add the sample to the centrifuge tube quantitatively, add the diluent to mix, and then add it to the reagent card with a pipette after mixing. After the sampling drive tube 100 of the present invention absorbs whole blood, it is directly inserted into the diluent storage chamber 126. After shaking, the entire device is inserted into the microfluidic chip 200, and the drive rod is rotated. No delicate operation is required throughout the process, and the fault tolerance rate is high.

[0066] 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 with the port at the other end of the liquid storage cylinder 120 through the sealing ring 134.

[0067] In one embodiment of the present invention, the first separation membrane 121 is a hydrophobic membrane, and the first separation membrane 121 separates the threaded cavity 124 from the microsphere storage chamber 125. The first separation membrane 121 allows air to pass through, but liquid cannot pass through. The second separation membrane 122 and the third separation membrane 123 are both metal membranes, and preferably, the second separation membrane 122 and the third separation membrane 123 are both metal aluminum membranes.

[0068] The present invention further provides a detection method of a microfluidic chip 200. The detection method is based on any of the above-mentioned microfluidic chips 200, and the detection method comprises: Step S100, quantitatively collecting samples through the sampling tube assembly 130, and inserting the sampling tube assembly 130 into the port at the other end of the liquid storage cylinder 120 to puncture the second separation membrane 122, so that the diluent flows into the microsphere storage chamber; Specifically, firstly, quantitatively collect samples through the capillary 133, tear open the third separation membrane 123, insert the sampling tube assembly 130 into the port at the other end of the liquid storage tube 120, insert one end of the capillary 133 into the diluent storage chamber 126, cover the sampling tube body 131, so that the capillary 133 pierces the second separation membrane 122, and releases the diluent pre-stored in the diluent storage chamber 126 (the diluent dosage is 100-1000 μL, and the type is phosphate solution), and the diluent is released and flows into the microsphere storage chamber.

[0069] Step S200, shaking the sampling driving tube 100 to mix the diluent, the fluorescent microspheres and the sample to form a reaction solution; Specifically, the sampling drive tube 100 is shaken upside down 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 the anti-human IgG secondary antibody to form a human IgG antibody-fluorescent microsphere complex, which is evenly distributed in the reaction solution with the remaining sample and microspheres that do not participate in the reaction.

[0070] Step S300, insert the outlet tube 132 into the sample injection port 220 and rotate the driving rod 110 to input the reaction liquid into the sample injection port 220, so that the reaction liquid enters the quantitative pool 241 and the sample waste liquid pool 246 respectively; Specifically, the outlet tube 132 is inserted into the sample addition port 220, and the driving rod 110 is rotated to drive the piston 112 to move slowly toward the sampling tube assembly 130, and the reaction liquid is added to the injection pool 244. The reaction liquid flows along the injection channel 240 to the three quantitative pools 241 under the drive of the piston until the three quantitative pools 241 are filled. The excess reaction liquid enters the sample waste liquid pool 246 through another injection channel 240 until the hydrophobic membrane in the waste liquid pool pore 278 is blocked, thereby completing the addition and separation of the reaction liquid.

[0071] Step S400, the reaction liquid in the quantitative pool 241 enters the corresponding reaction channel 250, so that the human IgG antibody-fluorescent microsphere complex in the reaction liquid reacts with the allergen antigen spot to form an antigen-human IgG antibody-fluorescent microsphere complex; Specifically, under the capillary action, the quantitative reaction liquid in the three quantitative pools 241 breaks 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 liquid reacts with the allergen antigen spots pre-embedded in the reaction flow channel 250 to form an antigen-human IgG antibody-fluorescent microsphere complex fixed on the chip. The remaining samples and fluorescent microspheres that did not participate in the reaction continue to flow with the reaction liquid. When the reaction liquid reaches the diversion port 263, due to the hydrophilic effect of the diversion port 263, the reaction liquid fills the diversion port 263 and is transferred to the transfer channel 262 of the lower layer, and enters the reaction waste liquid pool 261 along the transfer channel 262 under the capillary action to complete the immune reaction process.

[0072] Step S500, placing the microfluidic chip 200 into the instrument and squeezing the airbag, the airbag inputs airflow to the reaction channel 250 through the air channel 230, so as to transport the remaining reaction liquid in the reaction channel 250 to the corresponding reaction waste liquid pool 261; Specifically, after the capillary self-driven reaction is completed, the microfluidic chip 200 is inserted into a matching detection instrument, and the inclined surface structure 300 at the inlet of the instrument is used to squeeze the airbag, so that the gas in the airbag enters the reaction channel 250 along the airway 230, and the reaction liquid remaining in the reaction channel 250 is driven to the reaction waste liquid pool 261 at the end of the transfer channel 262, completing the waste discharge step. Figure 8 As shown, the inclined surface of the inclined surface structure 300 faces downward, and as the airbag enters the gap below the inclined surface, the gap becomes smaller, so that the airbag is gradually compressed. Step S600 , using an instrument to detect the antigen-human IgG antibody-fluorescent microsphere complex in the reaction channel 250 .

[0073] Specifically, the instrument uses a semiconductor laser to emit a laser of a specific wavelength to illuminate the reaction channel 250, and the antigen-human IgG antibody-fluorescent microsphere complex in 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 to form a two-dimensional image signal, and converts the image information to output a signal value to achieve the detection of different allergen indicators.

[0074] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions 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: Air supply device (210); A chip body, wherein the chip body is provided with a sample addition port (220), and an air channel (230), a plurality of injection channels (240), a plurality of reaction channels (250) and a reaction waste liquid holding chamber (260) are provided inside the chip body, wherein the plurality of injection channels (240) are connected to the sample addition port (220), the plurality of reaction channels (250) are connected to the plurality of injection channels (240) in a one-to-one correspondence, the plurality of reaction channels (250) are connected to the reaction waste liquid holding chamber (260), and the reaction channels (250) are provided with allergen antigen spots of different indicators; the air supply device (210) is connected to the reaction channel (250) through the air channel (230), and the air supply device (210) is used to input airflow to the reaction channel (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 holding chamber (260).

2. The microfluidic chip according to claim 1, characterized in that: The chip body comprises a bottom sealing layer (271), a reaction layer (272), a sampling layer (273) and an airway layer (274) which are stacked in sequence from bottom to top; the airway layer (274) is provided with an air inlet (275); both the airway layer (274) and the sampling layer (273) are provided with the sample addition port (220) and a plurality of quantitative pool air holes (276); the air supply device (210) is provided on a side of the airway layer (274) away from the sampling layer (273); the air supply device (210) is connected to the airway (230) through the air inlet (275); the plurality of quantitative pool air holes (276) are connected to the plurality of sampling channels (240) in a one-to-one correspondence; and the quantitative pool air holes (276) are provided with a hydrophobic membrane.

3. The microfluidic chip according to claim 2, characterized in that: The air supply device (210) comprises an air bag, the air bag being fixedly connected to a side of the airway layer (274) facing away from the sample inlet layer (273), the air bag being provided with an air outlet, the air outlet of the air bag being in communication with the air inlet hole (275).

4. The microfluidic chip according to claim 2, characterized in that: The injection channel (240) comprises: A plurality of quantitative pools (241) are arranged on a side of the reaction layer (272) facing the sample injection layer (273); A plurality of resistance flow channels (242) are arranged inside the reaction layer (272); A plurality of stop valves (243) are arranged inside the reaction layer (272), one end of the plurality of stop valves (243) is connected to the plurality of quantitative pools (241) in a one-to-one correspondence through the plurality of resistance flow channels (242), the other end of the plurality of stop valves (243) is connected to the plurality of reaction flow channels (250) in a one-to-one correspondence, and the gas channel (230) is connected to the other end of the plurality of stop valves (243).

5. The microfluidic chip according to claim 4, characterized in that: A sample injection pool (244), a sample injection channel (245) and a sample waste liquid pool (246) are further provided on the side of the reaction layer (272) facing the sample injection layer (273); the sample injection pool (244) is connected to the quantitative pool (241) via one of the sample injection channels (245); the sample injection pool (244) is connected to the sample addition port (220); the sample waste liquid pool (246) is connected to the quantitative pool (241) via another of the sample injection channels (245); the air channel layer (274) and the sample injection layer (273) are both provided with waste liquid pool pores (278); the waste liquid pool pores (278) are connected to the sample waste liquid pool (246); and the waste liquid pool pores (278) are provided with a hydrophobic membrane.

6. The microfluidic chip according to claim 5, characterized in that: The flow resistance of the resistance flow channel (242) is greater than the flow resistance of the injection flow channel (245).

7. The microfluidic chip according to any one of claims 2 to 6, characterized in that: The reaction waste liquid receiving chamber (260) comprises: A reaction waste liquid pool (261), arranged on a side of the reaction layer (272) away from the sample injection layer (273); A transfer channel (262) is arranged on a side of the reaction layer (272) away from the sample injection layer (273), and one end of the transfer channel (262) is connected to the reaction waste liquid pool (261); A flow guide port (263), wherein the flow guide port (263) penetrates the reaction layer (272) and is respectively connected to the other end of the transfer channel (262) and the reaction flow channel (250).

8. The microfluidic chip according to claim 7, characterized in that: The bottom sealing layer (271) is provided with a plurality of pores (264), the plurality of pores (264) are connected to the plurality of reaction waste liquid pools (261) in a one-to-one correspondence, and the pores (264) are provided with a hydrophobic membrane.

9. A device-free allergen multi-detection microfluidic chip, characterized in that: It comprises a sampling drive tube (100) and the microfluidic chip according to any one of claims 1 to 8.

10. A detection method for a microfluidic chip, the detection method being based on the microfluidic chip according to any one of claims 1 to 8, characterized in that: The detection method comprises: Quantitatively collecting samples through a sampling tube assembly (130), and inserting the sampling tube assembly (130) into a port at the other end of the liquid storage cylinder (120) to puncture the second separation membrane (122) so that the diluent flows into the microsphere storage chamber; Shaking the sampling drive tube (100) to mix the diluent, the fluorescent microspheres and the sample to form a reaction solution; Inserting the outlet tube (132) into the sample injection port (220) and rotating the driving rod (110) to input the reaction liquid into the sample injection port (220), so that the reaction liquid enters the quantitative pool (241) and the sample waste liquid pool (246) respectively; The reaction liquid in the quantitative pool (241) enters the corresponding reaction channel (250), so that the human IgG antibody-fluorescent microsphere complex in the reaction liquid reacts with the allergen antigen spot to form an antigen-human IgG antibody-fluorescent microsphere complex; The microfluidic chip is placed in an instrument and the air bag is squeezed, and the air bag inputs airflow to 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 flow channel (250) is detected using an instrument.

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