Microfluidic chip detection device and method

Abstract

The application relates to a micro-fluidic chip detection device and method. The detection device comprises a top layer, a channel layer, a reaction layer and a bottom layer. The channel layer is provided with a sample cavity, a dilution cavity, a mixing cavity and a labeling cavity which are sequentially lowered in vertical height. Liquid can sequentially flow from the sample cavity to the labeling cavity under the action of gravity, thereby ensuring the sufficiency of the reaction. The dilution liquid is pre-packaged in a dilution bag. After the sample is added, the dilution bag is punctured by pressing the pressing part, and the dilution liquid flows into the dilution cavity, thereby effectively preventing the dilution liquid from deteriorating due to long-term storage. The first mixing area is provided with stirring blades driven by a motor to rotate, thereby quickly and efficiently mixing the sample and the dilution liquid. The gas-permeable hole is provided with a hydrophobic gas-permeable membrane, which is beneficial to gas exchange. The detection cavity is provided with a drying cavity, which prevents the influence of long-term or humid environment storage on the detection result.

Description

Technical Field

[0001] The present application belongs to the field of microfluidic chip technology and relates to a microfluidic chip detection device and method. Background Art

[0002] Microfluidics, also known as microfluidic chip technology, integrates basic operational units such as sample preparation, reaction, separation, and detection in biological, chemical, and medical analysis processes onto a chip measuring a few square centimeters. Controlled fluids flow throughout the system, replacing various functions in conventional chemical or biological laboratories. Microfluidics offers advantages such as lightweight size, reduced sample and reagent volumes, rapid reaction speed, the ability to perform large-scale parallel processing, and disposable processing. Currently, microfluidics holds enormous potential for development and application in biomedical research.

[0003] Microfluidic chips are characterized by their integrated microchannel networks and numerous analytical functional components. Based on the principle of specific interactions between biomolecules, biochemical analysis processes can be integrated onto the chip surface, enabling high-throughput and rapid detection of biological components such as nucleic acids and proteins. Patent application number 201910239730.X discloses a microfluidic chip and an in vitro detection device containing the microfluidic chip. These devices utilize siphoning and external centrifugal forces to control the flow of samples into a quantitative chamber to achieve quantification of the test solution. However, centrifugation makes it difficult to control the amount of sample entering the detection chamber, and excessive centrifugal force can also cause loss of reactants or test substances, which can easily lead to experimental errors and affect the accuracy of test results. Summary of the Invention

[0004] The purpose of this application is to provide a microfluidic chip detection device and method, which uses gravity to achieve the flow of samples and can complete the entire reaction process without the help of external equipment.

[0005] According to one aspect of the present application, the present application provides a microfluidic chip detection device, which includes a top layer, a channel layer, a reaction layer, and a bottom layer connected by snap-fit;

[0006] The top layer is provided with an injection port and distributed air holes;

[0007] The vent holes include a first vent hole communicating with the dilution chamber, a second vent hole communicating with the mixing chamber, a third vent hole communicating with the marking chamber, a fourth vent hole communicating with the detection chamber, and a fifth vent hole communicating with the waste liquid chamber;

[0008] A hydrophobic breathable membrane is provided in the breathable hole.

[0009] In one embodiment of the present application, the channel layer is provided with a sample chamber, a dilution chamber, a mixing chamber, and a labeling chamber;

[0010] The injection port passes through the top layer and is connected to the sample cavity;

[0011] The sample chamber is in communication with the dilution chamber;

[0012] The cross-sectional shape of the sample chamber is a combination of a square and an inverted trapezoid, and the inverted trapezoid is closer to the dilution chamber than the square.

[0013] In one embodiment of the present application, a storage chamber is provided above the dilution chamber, and a flow hole is provided between the dilution chamber and the storage chamber.

[0014] In one embodiment of the present application, a pressing portion, a storage bag, a puncturing member, and a limiting portion are provided in the storage cavity. The lower end portion of the pressing portion is connected to the storage bag. The puncturing member is located at the bottom of the storage cavity and does not contact the storage bag in a free state. The cross-sectional shape of the lower end portion of the storage bag is an inverted trapezoid. In the pressed state, the lower end portion of the storage bag contacts the puncturing member.

[0015] In one embodiment of the present application, the pressing portion includes a pressing key, a pressing column, a stop bar, and a spring. The upper end of the pressing key is flush with the upper surface of the top layer, the lower end of the pressing key is connected to the upper end of the pressing column, the upper end of the stop bar contacts the lower end of the limiting portion in a free state, the spring is sleeved on the upper half of the pressing column, and the lower end of the spring is higher than the limiting portion.

[0016] In one embodiment of the present application, the mixing chamber includes a first mixing zone and a second mixing zone;

[0017] The first mixing zone is provided with a rotating shaft, a stirring blade, and a first pressure sensor. The rotating shaft is located at the center of the first mixing zone. The stirring blade is connected to the rotating shaft and is symmetrically arranged about the rotating shaft. The first pressure sensor is located at the inlet end of the first mixing zone.

[0018] A motor is provided in the channel layer, and an output shaft of the motor is connected to the rotating shaft;

[0019] The second mixing zone is a serpentine structure.

[0020] In one embodiment of the present application, the third vent hole passes through the top layer and communicates with the labeling cavity, and the labeling cavity is coated with a labeled antibody.

[0021] In one embodiment of the present application, a detection cavity is provided in the reaction layer, and the detection cavity includes a main channel, one end of the main channel is connected to the marking cavity, and the main channel extends to both sides to form a first detection area and a third detection area, respectively. The other end of the main channel extends away from the marking cavity and extends to both sides at the end of the main channel to form a third detection area and a fourth detection area.

[0022] The first detection area is provided with a first detection unit, a second detection unit, a third detection unit, a fourth detection unit, and a fifth detection unit in sequence;

[0023] The first detection unit is closer to the main channel than the second detection unit, the second detection unit is closer to the main channel than the third detection unit, the third detection unit is closer to the main channel than the fourth detection unit, and the fourth detection unit is closer to the main channel than the fifth detection unit.

[0024] In one embodiment of the present application, a waste liquid hole is provided at the lower end of the first detection unit, and the waste liquid hole is communicated with the waste liquid chamber;

[0025] The detection unit contains a detection card, the detection card contains a plurality of detection strips, and the detection strips are coated with different detection antibodies;

[0026] A drying chamber is provided below the first detection unit, a desiccant is provided in the drying chamber, a vent is provided in the drying chamber, and the drying chamber is connected to the detection chamber through the vent;

[0027] The bottom layer is provided with a waste liquid cavity, and the fifth vent is connected to the waste liquid cavity through the top layer, the reaction layer, and the detection layer;

[0028] The marking cavity is located lower than the sample cavity.

[0029] According to another aspect of the present application, the present application also provides a method for performing detection using the above-mentioned microfluidic chip detection device, comprising the following steps:

[0030] S1: Add the sample to the sample chamber through the sample inlet, wait for 1-30 seconds, press the button, the piercing element pierces the storage bag, and the diluent in the storage bag flows out and flows into the dilution chamber from the flow hole;

[0031] S2: The diluent drives the sample to flow into the first mixing zone. When the sample flows through the first pressure sensor, the first pressure sensor transmits a signal to the control center. The control center controls the motor to start working, and the motor drives the stirring blade to rotate, thereby mixing the sample and the diluent evenly.

[0032] S3: When the mixed liquid enters the second mixing zone and flows through the second pressure sensor, the second pressure sensor transmits a signal to the control center, and the control center controls the motor to stop working;

[0033] S4: The mixed solution is fully mixed in the second mixing zone and then enters the labeling chamber. In the labeling chamber, the analyte in the mixed solution combines with the labeled antibody to form an analyte-primary antibody complex;

[0034] S5: The complex enters the detection chamber under the action of gravity, flows into each branch channel through the main channel, and then flows into each detection unit, combines with the detection antibody on the detection strip, and the excess liquid flows into the waste liquid chamber.

[0035] The advantages of the present application are as follows: the sample chamber, dilution chamber, mixing chamber, and labeling chamber of the present application descend in sequence in vertical height, so that the liquid can flow sequentially from the sample chamber to the labeling chamber under the action of gravity, the end of the labeling chamber is connected with the detection chamber, the liquid flows downward from the channel layer to the reaction layer, the end of each detection unit in the reaction layer is connected with the waste liquid chamber on the bottom plate, the liquid flows downward from the reaction layer to the bottom layer, due to the height difference between the channel layer and the reaction layer, the liquid flowing into the reaction layer is difficult to flow back to the reaction layer, due to the height difference between the reaction layer and the bottom layer, the liquid flowing into the waste liquid chamber is difficult to flow back to the reaction layer, the liquid in each step can completely reach the next cavity, thereby ensuring the adequacy of the reaction and the accuracy of the detection results.

[0036] In the present application, the diluent is pre-packaged in a storage bag. After the sample is added, the storage bag is punctured by pressing the pressing portion downward, and the diluent flows into the dilution chamber, which effectively prevents the diluent from deteriorating due to long-term storage. At the same time, it also avoids the tedious operation process of preparing the diluent for immediate use and adding samples on site. The diluent can be released with just a light press, which greatly improves the detection efficiency. The first mixing area of ​​the present application is provided with a stirring blade. When the diluent drives the sample to flow into the first mixing area and flows through the first pressure sensor, the first pressure sensor transmits a signal to the control center, and the control center controls the motor to start working, and the motor drives the stirring blade to rotate, thereby quickly and efficiently mixing the sample and the diluent. The air vent of the present application is provided with a hydrophobic breathable membrane, which is conducive to gas exchange and prevents liquid from overflowing from the air vent. In addition, the present application is provided with multiple detection units, which can detect a large number of antigen substances. It only needs to coat the labeled antibody in the labeling cavity and the corresponding detection antibody in the detection cavity to detect a variety of antigen substances. A drying chamber is provided in the detection chamber, a desiccant is provided in the drying chamber, and a vent is provided in the drying chamber. The drying chamber and the detection chamber are connected through the vent. After the test strip is stored for a long time, the vent is prevented from affecting the test results of the microfluidic chip during long-term storage or storage in a humid environment, which helps to improve the stability of the microfluidic chip. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 Schematic diagram of the microfluidic chip of this application;

[0038] Figure 2 Schematic diagram of the upper surface (top layer) of the microfluidic chip;

[0039] Figure 3 Schematic diagram of the channel layer of the microfluidic chip;

[0040] Figure 4 Schematic diagram of the reaction layer of the microfluidic chip;

[0041] Figure 5 This is a schematic diagram of the bottom layer of the microfluidic chip;

[0042] Figure 6 Schematic diagram of the storage cavity;

[0043] Figure 7 Schematic diagram of the drying chamber;

[0044] In the figure: 1. Top layer; 2. Channel layer; 3. Reaction layer; 4. Bottom layer; 5. Inlet; 6. First air vent; 7. Second air vent; 8. Third air vent; 9. Fourth air vent; 10. Fifth air vent; 11. Sample chamber; 12. Dilution chamber; 13. Mixing chamber; 14. Marking chamber; 15. Storage chamber; 16. Test card; 17. Storage bag; 18. Piercing element; 19. Limiting part; 20. First mixing zone; 21. Second mixing zone; 22. Rotating shaft; 23. Stirring blade; 24. First pressure sensor; 25. Motor; 26. Second pressure sensor; 27. Main channel; 28 , first detection area; 29, second detection area; 30, third detection area; 31, fourth detection area; 32, first detection unit; 33, second detection unit; 34, third detection unit; 35, fourth detection unit; 36, fifth detection unit; 37, first branch channel; 38, second branch channel; 39, third branch channel; 40, fourth branch channel; 41, fifth branch channel; 42, sixth branch channel; 43, drying chamber; 44, vent; 45, press key; 46, press column; 47, gear bar; 48, spring; 49, waste liquid chamber; 50, flow hole; 51, detection card; 52, desiccant. DETAILED DESCRIPTION

[0045] The following examples are provided to illustrate the present invention in more detail, but they do not constitute a limitation of the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The reagents used in the following examples are commercially available common reagents unless otherwise specified.

[0046] like Figure 1As shown, this embodiment provides a microfluidic chip detection device, which includes a top layer 1, a channel layer 2, a reaction layer 3, and a bottom layer 4 from top to bottom. The top layer 1, the channel layer 2, the reaction layer 3, and the bottom layer 4 are connected by snapping.

[0047] In other embodiments of the present application, the connection method of the top layer 1, the channel layer 2, the reaction layer 3, and the bottom layer 4 is not limited, and can be a fixed connection, a movable connection, etc., preferably a movable connection, and more preferably a snap connection.

[0048] like Figure 2 As shown, the top layer 1 is provided with a sample inlet 5 and distributed air holes. The air holes include a first air hole 6 communicating with the dilution chamber 12, a second air hole 7 communicating with the mixing chamber 13, a third air hole 8 communicating with the labeling chamber 14, a fourth air hole 9 communicating with the detection chamber, and a fifth air hole 10 communicating with the waste liquid chamber 49.

[0049] Furthermore, there are multiple fourth ventilation holes 9 and multiple fifth ventilation holes 10 .

[0050] In other embodiments of the present application, the number of the first ventilation holes 6 , the second ventilation holes 7 , and the third ventilation holes 8 is not limited and can be multiple.

[0051] Furthermore, a hydrophobic breathable membrane is provided in the vent holes. The hydrophobic breathable membrane facilitates gas exchange and effectively prevents foreign matter from entering the microfluidic chip. It also prevents the liquid in the chip from overflowing from the vent holes and affecting the test results.

[0052] like Figure 3 As shown, the channel layer 2 is provided with a sample chamber 11, a dilution chamber 12, a mixing chamber 13, and a labeling chamber 14. The injection port 5 passes through the top layer 1 and is connected to the sample chamber 11; the sample chamber 11 is connected to the dilution chamber 12. In a specific embodiment, the cross-sectional shape of the sample chamber 11 is a combination of a square and an inverted trapezoid, and the inverted trapezoid is closer to the dilution chamber 12 to facilitate the sample to flow completely into the dilution chamber 12; in other embodiments of the present application, the cross-sectional shape of the sample chamber 11 is not limited.

[0053] The first vent hole 6 passes through the top layer 1 and is connected to the dilution chamber 12. A storage chamber 15 is provided above the dilution chamber 12. The storage chamber 15 is located between the first vent hole 6 and the injection port. A flow hole is provided between the dilution chamber 12 and the storage chamber 15. The dilution chamber 12 and the storage chamber 15 are connected through the flow hole. Figure 6As shown, a pressing portion 16, a storage bag 17, a piercing member 18, and a limiting portion 19 are provided in the storage chamber 15. The lower end of the pressing portion 16 is connected to the storage bag 17. The piercing member 18 is located at the bottom of the storage chamber 15 and does not contact the storage bag 17 in a free state (i.e., when the spring is not pressed, i.e., when the pressing key is not pressed), and the cross-section of the lower end of the storage bag 17 is an inverted trapezoid. In a pressed state (i.e., when the pressing key is pressed and the spring is compressed), the lower end of the storage bag 17 contacts the piercing member 18, which pierces the storage bag 17, causing the diluent in the storage bag 17 to flow out and into the dilution chamber 12 from the flow hole 50.

[0054] In a specific embodiment, the pressing portion 16 includes a pressing key 45, a pressing column 46, a stop bar 47, and a spring 48. The upper end of the pressing key 45 is flush with the upper surface of the top layer 1, and the lower end of the pressing key 45 is connected to the upper end of the pressing column 46. In the free state, the upper end of the stop bar 47 contacts the lower end of the limiting portion 19. The spring 48 is sleeved on the upper half of the pressing column 46, and the lower end of the spring 48 is higher than the limiting portion 19.

[0055] The mixing chamber 13 includes a first mixing zone and a second mixing zone. The second air vent 7 passes through the top layer 1 and is connected to the first mixing zone. The first mixing zone is located between the dilution chamber 12 and the second mixing zone. The first mixing zone is connected to the dilution chamber 12. The cross-sectional shape of the first mixing zone is circular. A rotating shaft 22, a stirring blade 23, and a first pressure sensor 24 are provided in the first mixing zone. The rotating shaft 22 is located in the center of the first mixing zone. The stirring blade 23 is connected to the rotating shaft 22. The stirring blade 23 is symmetrically arranged about the rotating shaft 22. The pressure sensor is located at the inlet end of the first mixing zone. Furthermore, a motor 25 is provided in the channel layer 2. The output shaft of the motor 25 is connected to the rotating shaft 22. When the mixture of the sample and the diluent flows into the first mixing zone and flows through the first pressure sensor 24, the first pressure sensor 24 transmits a signal to the control center (not shown in the figure), and the control center controls the motor 25 to start working.

[0056] In other embodiments of the present application, in order to achieve a better mixing effect, the shape of the stirring blades 23 can be wavy, spindle-shaped, etc., and the number of stirring blades 23 is not limited, and can be 2, 4, 6, 8, etc. The inner wall of the first mixing zone can be provided with protrusions of various shapes.

[0057] The second mixing zone is connected to the first mixing zone and is located between the first mixing zone and the marking chamber 14. The second mixing zone has a serpentine structure that helps further mix the sample and diluent. A second pressure sensor 26 is located at the outlet of the second mixing zone. When the sample and diluent mixture flows through the second pressure sensor 26, it transmits a signal to the control center, which in turn controls the motor 25 to stop.

[0058] The third air vent 8 passes through the top layer 1 and is connected to the labeling cavity 14. The labeling cavity 14 is coated with a labeled antibody. When the mixed liquid flows through the labeling cavity 14, the antigen in the mixed liquid combines with the labeled antibody to form a complex of the analyte-primary antibody. In a specific embodiment, the labeled antibody can be an antibody labeled with fluorescein, enzyme or biotin. The antibody can form a specific bond with a specific substance in the sample. There are multiple labeled antibodies. Furthermore, the number of labeled antibodies can be 5, 10, 20, 40, etc., so that the detection of multiple antigens can be achieved.

[0059] like Figure 4 As shown, the detection chamber is located in the reaction layer 3, and the detection chamber includes a main channel 27, a first detection area 28, a second detection area 29, a third detection area 30, and a fourth detection area 31. The fourth air vent 9 passes through the top layer 1 and the reaction layer 3 and is connected to the detection chamber. One end of the main channel 27 is connected to the marking chamber 14. The main channel 27 extends to both sides to form the first detection area 28 and the third detection area 30. The other end of the main channel 27 extends away from the direction of the marking chamber 14 and extends to both sides at the end of the main channel 27 to form the third detection area 30 and the fourth detection area 31. The first detection area 28 and the third detection area 30 are located on the same side, and the second detection area 29 and the fourth detection area 31 are located on the same side.

[0060] In a preferred embodiment of the present application, a first detection unit 32, a second detection unit 33, a third detection unit 34, a fourth detection unit 35, and a fifth detection unit 36 ​​are sequentially provided in the first detection area 28. The first detection unit 32 is closer to the main channel 27 relative to the second detection unit 33, the second detection unit 33 is closer to the main channel 27 relative to the third detection unit 34, the third detection unit 34 is closer to the main channel 27 relative to the fourth detection unit 35, and the fourth detection unit 35 is closer to the main channel 27 relative to the fifth detection unit 36.

[0061] In other embodiments of the present application, the number of detection units is not limited, and can be greater than five or less than five. They can be symmetrically arranged on both sides of the main channel 27 or individually arranged on one side of the main channel 27.

[0062] In this embodiment, the structure within the detection zone is described using first detection zone 28 as a representative example. It should be noted that the internal structures of the other detection zones, including second detection zone 29, third detection zone 30, etc., are identical to those of first detection zone 28. First detection zone 28 also includes a first branch channel 37, a second branch channel 38, a third branch channel 39, a fourth branch channel 40, a fifth branch channel 41, and a sixth branch channel 42, which are interconnected. One end of first branch channel 37 is connected to main channel 27, and the other end of first branch channel 37 is connected to one end of second branch channel 38. First branch channel 37 and second branch channel 38 form an L-shaped structure. The other end of second branch channel 38 is connected to one end of third branch channel 39, and the other end of third branch channel 39 is connected to one end of fourth branch channel 40. The other end of fourth branch channel 40 is connected to one end of fifth branch channel 41, and the other end of fifth branch channel 41 is connected to sixth branch channel 42.

[0063] Among them, the third branch channel 39 is farther away from the line A extending from the end of the main channel 27A to both sides of the width direction of the microfluidic chip than the second branch channel 38. The line A here is only for illustrating the distribution of each branch channel. The fourth branch channel 40 is farther away from the line A than the third branch channel 39, the fifth branch channel 41 is farther away from the line A than the fourth branch channel 40, and the sixth branch channel 42 is farther away from the line A than the fifth branch channel 41.

[0064] Furthermore, the distance from the third branch channel 39 to the second branch channel 38 is d, the distance from the fourth branch channel 40 to the third branch channel 39 is also d, the distance from the fifth branch channel 41 to the fourth branch channel 40 is also d, and the distance from the sixth branch channel 42 to the fifth branch channel 41 is also d.

[0065] In other embodiments of the present application, the distances between adjacent branch channels may be unequal.

[0066] In this embodiment, the second branch channel 38 is connected to the first detection unit 32, the upper end of the first detection unit 32 is connected to the second branch channel 38, and the lower end of the first detection unit 32 is provided with a waste liquid hole, which is connected to the waste liquid chamber 49.

[0067] In this embodiment, each detection unit contains a detection card containing multiple detection strips. Each of the detection strips is coated with different detection antibodies (secondary antibodies), and the detection antibodies correspond to the type and quantity of the labeled antibodies. In the detection zone, the sample-to-be-tested-primary-antibody complex forms a ternary complex of sample-to-be-tested-primary-antibody-secondary-antibody with the detection antibodies on the detection strips.

[0068] Furthermore, in this embodiment, there is one detection strip, and each detection strip is coated with one detection antibody, that is, each detection area can detect 5 antigens, and the detection cavity can detect 20 antigens.

[0069] In another embodiment of the present application, there are two detection strips, each of which is coated with a detection antibody, that is, each detection area can detect 10 antigens, and the detection cavity can detect 40 antigens.

[0070] In another embodiment of the present application, there are two detection strips, each of which is coated with a detection antibody, that is, each detection area can detect 15 antigens, and the detection cavity can detect 60 antigens.

[0071] In one embodiment, a quality control marker is coated within the marker cavity 14, and a quality control line is provided on the detection strip, which is coated with a quality control antibody. The quality control marker specifically binds to the quality control antibody to form a quality control marker-quality control antibody complex. The validity of the test results of the microfluidic chip is determined by detecting the quality control marker-quality control antibody complex.

[0072] Furthermore, if Figure 7 As shown, a drying chamber 43 is provided below the first detection unit 32, and a desiccant is provided in the drying chamber 43. The drying chamber 43 is located directly below the detection card, and a vent 44 is provided in the drying chamber 43. The drying chamber 43 is connected to the detection chamber through the vent 44. After the detection strip is stored for a long time, moisture is prevented from affecting the detection results of the microfluidic chip, which helps to improve the stability of the microfluidic chip.

[0073] Furthermore, a drying chamber 43 is provided below each detection unit.

[0074] like Figure 5 As shown, the waste liquid chamber 49 is located in the bottom layer 4, and the fifth vent 10 is connected to the waste liquid chamber through the top layer 1, the reaction layer 3, and the detection layer. After the mixed liquid combines with the detection antibody, the excess mixed liquid flows into the waste liquid chamber.

[0075] In this embodiment, the position of the marking chamber 14 is slightly lower than the sample chamber 11 , that is, under the action of gravity, the liquid slowly flows from the sample chamber 11 to the marking chamber 14 .

[0076] This application also includes a method for performing detection using the above-mentioned microfluidic chip detection device, comprising the following steps:

[0077] S1: Add sample to the sample chamber through the sample inlet 5, wait 1-30 seconds, press the button, the piercing member 18 pierces the storage bag 17, and the diluent in the storage bag 17 flows out and flows into the dilution chamber 12 through the flow hole;

[0078] S2: The diluent drives the sample to flow into the first mixing zone. When the sample flows through the first pressure sensor 24, the first pressure sensor 24 transmits a signal to the control center, which controls the motor 25 to start working. The motor 25 drives the stirring blade 23 to rotate, thereby mixing the sample and the diluent evenly.

[0079] S3: When the mixed liquid enters the second mixing zone and flows through the second pressure sensor 26, the second pressure sensor 26 transmits a signal to the control center, and the control center controls the motor 25 to stop working;

[0080] S4: The mixed solution is fully mixed in the second mixing zone and then enters the labeling chamber 14. In the labeling chamber 14, the analyte in the mixed solution combines with the labeled antibody to form an analyte-primary antibody complex;

[0081] S5: The complex enters the detection chamber under the action of gravity, flows into each branch channel through the main channel 27, and then flows into each detection unit. The analyte-primary antibody complex in the mixed liquid forms a ternary complex of analyte-primary antibody-secondary antibody with the detection antibody. The amount of the ternary complex in the detection area is detected to calculate the content of the analyte in the sample. The mixed liquid that is not bound to the detection antibody flows into the waste liquid chamber. The quality control marker specifically binds to the quality control antibody on the quality control line to form a quality control marker-quality control antibody complex. The effectiveness of the microfluidic chip is determined by detecting the quality control marker-quality control antibody complex in the detection chamber.

[0082] The sample chamber, dilution chamber, mixing chamber and marking chamber of the present application descend in sequence in vertical height, so that the liquid can flow sequentially from the sample chamber to the marking chamber under the action of gravity, the end of the marking chamber is connected with the detection chamber, the liquid flows downward from the channel layer to the reaction layer, and the end of each detection unit in the reaction layer is connected with the waste liquid chamber on the bottom plate, and the liquid flows downward from the reaction layer to the bottom layer. Due to the height difference between the channel layer and the reaction layer, the liquid flowing into the reaction layer is difficult to flow back to the reaction layer. Due to the height difference between the reaction layer and the bottom layer, the liquid flowing into the waste liquid chamber is difficult to flow back to the reaction layer. The liquid in each step can completely reach the next cavity, thereby ensuring the adequacy of the reaction and the accuracy of the test results.

[0083] In the present application, the dilution is pre-packaged in a dilution bag. After the sample is added, the dilution bag is punctured by pressing the pressing part, and the dilution liquid flows into the dilution chamber, which effectively prevents the dilution liquid from being deteriorated due to long-term storage. At the same time, it also avoids the tedious operation process of preparing the dilution liquid for immediate use and adding samples on site. The dilution liquid can be released with just a light press, which greatly improves the detection efficiency. In the present application, a stirring blade is provided in the first mixing area. When the dilution liquid drives the sample to flow into the first mixing area and flows through the first pressure sensor, the first pressure sensor transmits a signal to the control center. The control center controls the motor to start working, and the motor drives the stirring blade to rotate, thereby quickly and efficiently mixing the sample and the dilution liquid. A hydrophobic breathable membrane is provided in the air vent of the present application, which is conducive to gas exchange and prevents liquid from overflowing from the air vent. In addition, the present application is provided with multiple detection units, which can detect a large number of antigen substances. It only needs to coat the labeled antibody in the labeling cavity and the corresponding detection antibody in the detection cavity to detect a variety of antigen substances. A drying chamber is provided in the detection chamber, a desiccant is provided in the drying chamber, and a vent is provided in the drying chamber. The drying chamber and the detection chamber are connected through the vent. After the test strip is stored for a long time, the vent is prevented from affecting the test results of the microfluidic chip during long-term storage or storage in a humid environment, which helps to improve the stability of the microfluidic chip.

[0084] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0085] The above-described embodiments merely represent several implementation methods of the present application. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person of ordinary skill in the art may make various modifications and improvements without departing from the spirit of the present application, and such modifications and improvements are all within the scope of protection of the present application. Therefore, the scope of protection of the present application shall be determined by the appended claims.

Claims

1. A microfluidic chip detection device, characterized in that: It includes a top layer, a channel layer, a reaction layer, and a bottom layer connected by snap-fitting; the top layer is provided with an injection port and distributed air holes; the channel layer is provided with a sample chamber, a dilution chamber, a mixing chamber, and a labeling chamber; the reaction layer is provided with a detection chamber; and the bottom layer is provided with a waste liquid chamber. The vent holes include a first vent hole communicating with the dilution chamber, a second vent hole communicating with the mixing chamber, a third vent hole communicating with the marking chamber, a fourth vent hole communicating with the detection chamber, and a fifth vent hole communicating with the waste liquid chamber; a hydrophobic breathable membrane is provided in the vent holes; The injection port passes through the top layer and is connected to the sample chamber; the sample chamber is connected to the dilution chamber; the cross-sectional shape of the sample chamber is a combination of a square and an inverted trapezoid, and the inverted trapezoid is closer to the dilution chamber than the square; A storage chamber is provided above the dilution chamber, and a flow hole is provided between the dilution chamber and the storage chamber; The storage cavity is provided with a pressing portion, a storage bag, a piercing member, and a limiting portion. The lower end of the pressing portion is connected to the storage bag. The piercing member is located at the bottom of the storage cavity and does not contact the storage bag in a free state. The cross-section of the lower end of the storage bag is an inverted trapezoid. In the pressed state, the lower end of the storage bag contacts the piercing member. The pressing portion includes a pressing key, a pressing column, a stop bar, and a spring. The upper end of the pressing key is flush with the upper surface of the top layer, the lower end of the pressing key is connected to the upper end of the pressing column, the upper end of the stop bar contacts the lower end of the limiting portion in a free state, the spring is sleeved on the upper half of the pressing column, and the lower end of the spring is higher than the limiting portion. The mixing chamber includes a first mixing zone and a second mixing zone; a rotating shaft, a stirring blade, and a first pressure sensor are provided in the first mixing zone, the rotating shaft is located in the center of the first mixing zone, the stirring blade is connected to the rotating shaft, and the stirring blade is symmetrically arranged about the rotating shaft, and the first pressure sensor is located at the inlet end of the first mixing zone; a motor is provided in the channel layer, and the output shaft of the motor is connected to the rotating shaft; the second mixing zone is a serpentine structure.

2. The microfluidic chip detection device according to claim 1, characterized in that: The third vent hole passes through the top layer and is communicated with the marking cavity, and the marking cavity is coated with a marked antibody.

3. The microfluidic chip detection device according to claim 1, characterized in that: The detection chamber includes a main channel, one end of which is connected to the marking chamber, and the main channel extends to both sides to form a first detection area and a third detection area, respectively. The other end of the main channel extends away from the marking chamber and extends to both sides at the end of the main channel to form a third detection area and a fourth detection area. The first detection area is provided with a first detection unit, a second detection unit, a third detection unit, a fourth detection unit, and a fifth detection unit in sequence; The first detection unit is closer to the main channel than the second detection unit, the second detection unit is closer to the main channel than the third detection unit, the third detection unit is closer to the main channel than the fourth detection unit, and the fourth detection unit is closer to the main channel than the fifth detection unit.

4. The microfluidic chip detection device according to claim 3, characterized in that: A waste liquid hole is provided at the lower end of the first detection unit, and the waste liquid hole is connected to the waste liquid cavity; The detection unit contains a detection card, which contains multiple detection strips, and the detection strips are coated with different detection antibodies; A drying chamber is provided below the first detection unit, a desiccant is provided in the drying chamber, a vent is provided in the drying chamber, and the drying chamber is connected to the detection chamber through the vent; The fifth vent is connected to the waste liquid chamber through the top layer, the reaction layer, and the detection layer; The marking cavity is located lower than the sample cavity.

5. A method for detecting using the microfluidic chip detection device as claimed in claim 4, comprising the following steps: S1: Add the sample to the sample chamber through the sample inlet, wait for 1-30 seconds, press the button, the piercing element pierces the storage bag, and the diluent in the storage bag flows out and flows into the dilution chamber from the flow hole; S2: The diluent drives the sample to flow into the first mixing zone. When the sample flows through the first pressure sensor, the first pressure sensor transmits a signal to the control center. The control center controls the motor to start working, and the motor drives the stirring blade to rotate, thereby mixing the sample and the diluent evenly. S3: When the mixed liquid enters the second mixing zone and flows through the second pressure sensor, the second pressure sensor transmits a signal to the control center, and the control center controls the motor to stop working; S4: The mixed solution is fully mixed in the second mixing zone and then enters the labeling chamber. In the labeling chamber, the analyte in the mixed solution combines with the labeled antibody to form an analyte-primary antibody complex; S5: The complex enters the detection chamber under the action of gravity, flows into each branch channel through the main channel, and then flows into each detection unit, combines with the detection antibody on the detection strip, and the excess liquid flows into the waste liquid chamber.

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