A magnetic particle luminescent double-layer microfluidic chip and detection system

Abstract

The present invention belongs to the technical field of microfluidic chip luminescent immunoassay, and particularly relates to a magnetic particle luminescent double-layer microfluidic chip and a detection system. The chip includes a top plate and a bottom plate, the top plate includes a sample adding part, a labeled ligand storage part and a sample mixing area, and the sample mixing area is respectively connected to the sample adding part and the labeled ligand storage part; the bottom plate includes a guide area, a magnetic particle coating part, a cleaning area, a detection area and a cleaning liquid storage part, and the guide area is provided with a groove whose height is lower than the bottom wall of the magnetic particle coating part and a guide part provided on the groove and connected to the magnetic particle coating part. The sample enters from the sample adding part, mixes with the labeled ligand in the sample mixing area, and then enters the groove of the guide area. The sample in the groove can only be sucked away by capillary action, and due to the action of the truncation groove and the blocking part, the sample can only enter the magnetic particle coating part from the guide part, and then mix and react with the magnetic particle ligand, and after cleaning in the cleaning area, luminescent detection is realized in the detection area.

Description

Technical Field

[0001] The invention belongs to the technical field of microfluidic chip luminescent immunoassay, and in particular relates to a magnetic particle luminescent double-layer microfluidic chip and a detection system. Background Art

[0002] At present, there are two main development trends of in vitro diagnosis (IVD): one is automation and integration, that is, using the fully automated, highly sensitive large-scale instruments and equipment of the central laboratory of large hospitals to achieve high-precision disease analysis and diagnosis; the other is miniaturization and bedside, that is, through small and simple handheld equipment, to achieve on-site rapid analysis and diagnosis. However, small hospitals are short of funds and have small sample sizes, and are not suitable for purchasing expensive large equipment. Therefore, the rapid detection equipment used by most hospitals at this stage is mainly test strips and their supporting equipment, but test strips can only achieve qualitative or semi-quantitative detection, with low detection sensitivity, poor specificity, poor repeatability, and obvious interference. Due to China's large population, aggravated aging, and a sharp increase in the incidence rate, relying solely on large hospitals is overwhelmed. Therefore, it has become extremely urgent to develop rapid detection methods and equipment that are easy to operate, highly sensitive, repeatable, and quantitatively accurate.

[0003] Microfluidic chip technology integrates the basic operation units such as sample preparation, reaction, separation, and detection in the biological, chemical, and medical analysis process into a micron-scale chip to automatically complete the entire analysis process. Due to its huge potential in the fields of biology, chemistry, and medicine, it has developed into a multidisciplinary research field such as biology, chemistry, medicine, fluids, materials, and machinery, and has been applied to biomedical research, biochemical testing, forensic identification, and other fields. However, the existing microfluidic chip is provided with a top plate and a bottom plate. When the sample flows from the top plate to the bottom plate, since the channels of the bottom plate are at the same level, the sample will automatically flow backward along the channel due to gravity. In some detection projects, only a small amount of the sample needs to be taken to complete the detection and analysis. If the sample flows from the top plate to the bottom plate and flows directly in the bottom plate, it will greatly affect the final detection result and cause errors in the detection result. In addition, the sample flows along the channel of the bottom plate, and the flow path of the sample cannot be limited. It is impossible to ensure that the sample flowing to the detection area is completely filtered, which further affects the final detection result. Summary of the invention

[0004] The embodiment of the present invention provides a magnetic particle luminescent double-layer microfluidic chip and a detection system, aiming to solve the problem that in the existing microfluidic chip, after the sample flows from the top plate to the bottom plate, it flows directly in the bottom plate and the flow path of the sample cannot be limited.

[0005] The embodiment of the present invention is implemented as follows: a magnetic particle luminescent double-layer microfluidic chip is provided, the chip comprising: a top plate, the top plate comprising a sample loading section, a labeled ligand storage section and a sample mixing area, the labeled ligand storage section is provided with a labeled ligand, and the sample mixing area is interconnected with the sample loading section and the labeled ligand storage section respectively; a bottom plate arranged on the top plate, the bottom plate comprising a guide area interconnected with the sample mixing area, a magnetic particle coating section interconnected with the guide area, a cleaning area interconnected with the magnetic particle coating section, a detection area interconnected with the cleaning area, and a cleaning liquid storage section interconnected with the cleaning area, the guide area is provided with a groove whose height is lower than the bottom wall of the magnetic particle coating section, a guide section arranged on the groove and connected to the magnetic particle coating section, a truncation groove arranged below the front end of the guide section, and a blocking section arranged on the guide section, the magnetic particle coating section is provided with a magnetic particle ligand solution, and the cleaning liquid storage section is provided with a cleaning liquid.

[0006] Furthermore, the top plate also includes an air pump interconnected with the sample adding part.

[0007] Furthermore, the top plate is provided with elastic members at corresponding positions of the air pump and the sample mixing area.

[0008] Furthermore, a porous elastic member is provided inside the air pump.

[0009] Furthermore, the sample adding part includes a sample adding port and a cover for opening or closing the sample adding port, and the sample adding part also includes a rubber ring arranged on the sample adding port.

[0010] Furthermore, the top plate and the bottom plate are both provided with limiting notches at positions corresponding to each other.

[0011] Furthermore, the top plate is provided with a first buckle or a first slot, and the bottom plate is provided with a second slot or a second buckle, and the top plate and the bottom plate are buckled with each other through the mutual cooperation between the first buckle and the second slot, or through the mutual cooperation between the first slot and the second buckle.

[0012] Furthermore, a single-sided adhesive material is provided on a portion or the entire area of ​​at least one side of the bottom plate.

[0013] Furthermore, a product label is provided on the surface of the top plate or the bottom plate, and a QR code label is provided on the surface of the top plate or the bottom plate.

[0014] Furthermore, the top plate is provided with magnetic absorption holes on corresponding connecting tracks with the magnetic particle coating part, the cleaning area and the detection area.

[0015] Furthermore, the bottom plate also includes a waste liquid pool interconnected with the cleaning area.

[0016] Furthermore, the bottom plate also includes a luminescent liquid storage portion interconnected with the detection area, and the luminescent liquid is provided inside the luminescent liquid storage portion.

[0017] Furthermore, the top plate is provided with a cleaning liquid evacuation hole and a luminescent liquid evacuation hole at corresponding positions of the cleaning liquid storage part and the luminescent liquid storage part.

[0018] Furthermore, a fluorescent liquid is provided inside the labeled ligand storage unit.

[0019] The present invention also provides a magnetic particle luminescent double-layer microfluidic detection system, which includes: the magnetic particle luminescent double-layer microfluidic chip as described above; a magnet unit for driving the magnetic particles in the magnetic particle ligand solution to move; an extrusion unit for squeezing the labeled ligand storage part and the cleaning liquid storage part to allow the labeled ligand and the cleaning liquid to flow out; and a detection unit for detecting the luminescent signal in the detection area.

[0020] The beneficial effect of the present invention lies in that, compared with the prior art, the present invention designs a magnetic particle luminescent double-layer microfluidic chip and a detection system. The sample enters from the sample adding part, mixes with the labeled ligand in the sample mixing area, and then enters the groove in the guide area. Since the groove is lower than the bottom wall of the magnetic particle coating part, the sample in the groove can only be sucked away by capillary action, and due to the action of the cut-off groove and the blocking part, the sample can only enter the magnetic particle coating part from the guide part, and then fully mix and react with the magnetic particle ligand, and after being cleaned with the cleaning liquid in the cleaning area, luminescent detection is realized in the detection area. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 1 is an exploded schematic diagram of a magnetic particle luminescent double-layer microfluidic chip provided in an embodiment of the present invention;

[0022] Figure 2 yes Figure 1 A partial enlarged view of

[0023] Figure 3 This is another exploded schematic diagram of the magnetic particle luminescent double-layer microfluidic chip provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0025] The present invention designs a magnetic particle luminescent double-layer microfluidic chip and a detection system. The sample enters from the sample adding part 11, mixes with the labeled ligand in the sample mixing area 13, and then enters the groove 211 of the guide area 21. Since the groove 211 is lower than the bottom wall of the magnetic particle coating part 22, the sample in the groove 211 can be sucked away by capillary action. The sample enters the magnetic particle coating part 22 from the guide part 212, and then fully mixes and reacts with the magnetic particle ligand. After being cleaned with the cleaning liquid in the cleaning area 23, luminescent detection is realized in the detection area 24.

[0026] Embodiment 1

[0027] refer to Figure 1 and Figure 2 The first embodiment of the present invention provides a magnetic particle luminescent double-layer microfluidic chip. The magnetic particle luminescent double-layer microfluidic chip includes a top plate 1 and a bottom plate 2 arranged on the top plate 1 .

[0028] The top plate 1 includes a sample adding part 11, a labeled ligand storage part 12 and a sample mixing area 13. The labeled ligand storage part 12 is provided with a labeled ligand inside, and the sample mixing area 13 is communicated with the sample adding part 11 and the labeled ligand storage part 12 respectively.

[0029] The bottom plate 2 includes a flow guide area 21 interconnected with the sample mixing area 13, a magnetic particle coating part 22 interconnected with the flow guide area 21, a cleaning area 23 interconnected with the magnetic particle coating part 22, a detection area 24 interconnected with the cleaning area 23, and a cleaning liquid storage part 25 interconnected with the cleaning area 23. The flow guide area 21 is provided with a groove 211 whose height is lower than the bottom wall of the magnetic particle coating part 22, a flow guide part 212 arranged on the groove 211 and connected to the magnetic particle coating part 22, a truncation groove 213 arranged below the front end of the flow guide part 212, and a blocking part 214 arranged on the flow guide part 212. The flow guide part 212 can be a blood filter membrane, and the blocking part 214 can be plastic paper. The bottom wall of the truncation groove 213 is also provided with a blood filter membrane. The height of the truncation groove 213 is lower than the height of the groove 211. The magnetic particle coating part 22 is provided with a magnetic particle ligand solution, and the cleaning liquid storage part 25 is provided with a cleaning liquid.

[0030] For example, if the sample is a whole blood sample, the sample to be tested can be placed in the sample adding part 11, and the sample enters the sample mixing area 13 through the sample adding part 11. At this time, the labeled ligand inside the labeled ligand storage part 12 also enters the sample mixing area 13, and the sample and the labeled ligand are mixed in the sample mixing area 13, and then enter the guide part 212 of the guide area 21 from the sample mixing area 13. After passing through the guide area 21, the plasma and blood cells in the sample are separated, and the plasma enters the magnetic particle coating part 22 from the guide part 212, while the blood cells remain in the guide area 21. Since the groove 211 of the guide area 21 is lower than the bottom wall of the magnetic particle coating portion 22, similarly, the height of the guide portion 212 is also lower than the bottom wall of the magnetic particle coating portion 22. After the sample enters the guide area 21, it will not automatically enter the magnetic particle coating portion 22 from the guide portion 212 due to gravity, but the sample will be sucked away from the guide portion 212 by capillary action. In this way, a smaller volume of sample that can meet the detection requirements can be sucked away from a larger volume of sample, thereby avoiding the large volume of the sample affecting the detection result. Furthermore, when the sample enters the flow guide area 21, due to the action of the cut-off groove 213 and the blocking part 214, the sample can only flow over the blocking part 214, and the sample that does not flow through the flow guide part 212 will fall into the cut-off groove 213, so that the sample can only flow from the flow guide part 212 into the capillary channel of the magnetic particle coated part 22, limiting the flow area of ​​the sample flowing into the magnetic particle coated part 22, and will not flow out of the flow guide part 212, causing the test sample to flow into the capillary channel of the magnetic particle coated part 22 from other areas, so that the filtering effect is better and the test result is more accurate. It is worth mentioning that if the test sample is whole blood, the red blood cells will not flow over the capillary channel of the magnetic particle coated part 22 from the front end of the flow guide part 212, but will be subjected to the capillary action of the capillary channel, and the plasma after adsorption and filtration will enter the magnetic particle coated part 22.

[0031] After the sample after the flow is directed reaches the corresponding position of the magnetic particle coating part 22, the analyte in the sample reacts with the magnetic particle ligand solution, the external magnet collects the magnetic particles in the magnetic particle ligand solution, and the sample after the reaction enters the washing area 23, and the external magnet also drives the magnetic particles into the washing area 23. At this time, the washing liquid in the washing liquid storage part 25 is released, and the washing liquid enters the washing area 23 to wash the magnetic particles in the sample, and the sample then enters the detection area 24 from the washing area 23, and the external magnet also drives the magnetic particles into the detection area 24, thereby realizing the quantitative detection of the analyte in the sample.

[0032] The blood filter membrane is pre-set in the flow guide area 21, wherein the blood filter membrane can separate the liquid from the cells through physical pores or biological / chemical reagents, thereby separating the plasma from the red blood cells, and the plasma flows to the magnetic particle coating part 22, while the red blood cells stay on the blood filter membrane, thereby reducing the interference of the red blood cells on the test results. The biological / chemical reagents include coagulants, etc., which can connect the red blood cells, form clots, and increase the size. The red blood cells after the increase in size are more easily blocked by the mesh structure of the blood filter membrane, thereby more effectively reducing the interference of the red blood cells on the experimental results.

[0033] The cleaning solution is pre-stored in the cleaning solution storage unit 25, and the cleaning solution is used to clean the magnetic beads and remove non-specifically adsorbed analytes, luminescent markers and other substances that affect the test results. The cleaning solution mainly contains buffer reagents, proteins and surfactants, wherein the buffer reagents include but are not limited to borate, phosphate, Tris-HCl and acetate, etc., and the pH range of the cleaning solution is 6.0-10.0. The proteins include but are not limited to bovine serum albumin, casein, etc. The surfactant includes but is not limited to Tween 20, Tween 80, Triton X-100, polyethylene glycol and polyvinyl pyrrolidone, etc. Preferably, in this embodiment, the cleaning solution used is a pH7.0 Tris-HCl buffer containing bovine serum albumin, Tween 2 0 and Proclin 300.

[0034] In this embodiment, the labeled ligand storage part 12, the magnetic particle coating part 22 and the cleaning liquid storage part 25 are sealed cavities, and the sealing material used is an elastic material or a high barrier film, specifically glass, plastic, rubber, aluminum foil or a high barrier film, wherein the sealing material can be composed of the same material or a combination of multiple materials. Under physical extrusion, the labeled ligand storage part 12, the magnetic particle coating part 22 and the cleaning liquid storage part 25 can be partially ruptured, thereby releasing the stored materials.

[0035] Embodiment 2

[0036] refer to Figure 1 On the basis of the first embodiment, the top plate 1 of the fourth embodiment is further provided with an air pump 14 interconnected with the sample adding part 11. The air pump 14 is an air bag built into the top plate 1. By repeatedly squeezing or releasing the air bag, the air in the air bag repeatedly enters and exits the sample adding part 11 and the sample mixing area 13, thereby driving the liquid in the top plate 1 to flow.

[0037] The air pump 14 is used to absorb or squeeze the air in the sample mixing area 13, so that the sample and the labeled ligand flow to the sample mixing area 13, and by repeatedly absorbing or squeezing the air in the top plate 1, the sample and the labeled ligand are fully mixed in the sample mixing area 13, and after mixing, the sample is driven from the sample mixing area 13 into the guide area 21.

[0038] It should be noted that since the air pump 14 needs to work in a sealed environment, in order to seal the inside of the chip, it is necessary to close the cover after adding the sample to the sample port to seal the inside of the chip and ensure the driving effect of the air pump 14.

[0039] Embodiment 3

[0040] refer to Figure 1 On the basis of the second embodiment, the top plate 1 of the third embodiment is provided with an elastic member at the air pump 14 and the sample mixing area 13. The elastic member can be optionally glued to one side of the top plate 1, and the one side of the top plate 1 is adjacent to the bottom plate 2. The elastic member can provide elasticity for the air pump 14 and the sample mixing area 13.

[0041] Embodiment 4

[0042] refer to Figure 1 On the basis of the second embodiment, the air pump 14 of the fourth embodiment is provided with a porous elastic member inside, and the porous elastic member can be a sponge. The air pump 14 needs to be pressed repeatedly. After pressing the air pump 14, the pressing force is removed, and the porous elastic member can provide a rebound force for the pump surface of the air pump 14, so that the pump surface returns to a tight state. The porous elastic member is provided with a plurality of orifices, which can enable the porous elastic member to reduce the volume inside the air pump 14, and help the air pump 14 to be further miniaturized.

[0043] Embodiment 5

[0044] refer to Figure 1 On the basis of the first embodiment, the sample adding part 11 of the fifth embodiment includes a sample adding port and a cover for opening or closing the sample adding port.

[0045] When the cover is opened, a sample can be added to the sample adding port from outside, and after the sample is added, the cover is closed to seal the sample adding port.

[0046] Specifically, the cover is provided with a first clamp or a first clamp hole, and a second clamp hole or a second clamp is provided at a position adjacent to the sample adding port, and the cover closes the sample adding port through the cooperation between the first clamp and the second clamp hole, or the cooperation between the first clamp hole and the second clamp. In addition, the cover is also provided with a sealing member adapted to the sample adding port, and when the cover is closed, the sealing member is simultaneously inserted into the sample adding port to prevent the sample from leaking out of the sample adding port.

[0047] Furthermore, the sample adding part 11 further comprises a rubber ring disposed on the sample adding port. Since the sample is often added from the outside through a pipette tip, the rubber ring is elastic and helps to seal with the pipette tip, so that the sample can be injected from the sample adding port more smoothly.

[0048] Embodiment 6

[0049] On the basis of the first embodiment, the top plate 1 and the bottom plate 2 of the sixth embodiment are provided with limiting notches at corresponding positions. The limiting notches are notches provided at one end of the top plate 1 and the bottom plate 2, which are not shown in the figure. Since the magnetic particle luminescent double-layer microfluidic chip is in the detection process, in order to avoid the displacement of the magnetic particle luminescent double-layer microfluidic chip, it is necessary to perform limiting operation on the magnetic particle luminescent double-layer microfluidic chip. The limiting notch cooperates with the external limiting structure to firmly fix the chip during the detection process to avoid its displacement, thereby ensuring the smooth progress of the detection.

[0050] Embodiment 7

[0051] refer to Figure 1 On the basis of the first embodiment, the top plate 1 of the seventh embodiment is provided with a first buckle or a first slot, and the bottom plate 2 is provided with a second slot or a second buckle. The top plate 1 and the bottom plate 2 are buckled together by the cooperation between the first buckle and the second slot, or by the cooperation between the first slot and the second buckle. Through the above-mentioned buckling method, the top plate 1 and the bottom plate 2 can be detachably arranged, which is convenient for operators to check or replace. After buckling, it can ensure that the top plate 1 and the bottom plate 2 are firmly fixed to each other.

[0052] Of course, the top plate 1 and the bottom plate 2 may also be combined with each other in other ways, which will not be described in detail here.

[0053] Embodiment 8

[0054] refer to Figure 3 On the basis of the first embodiment, in the eighth embodiment, at least one side of the bottom plate 2 is partially or entirely provided with a single-sided adhesive material 3, and the single-sided adhesive material 3 can be a single-sided tape. Preferably, both sides of the bottom plate 2 are provided with a single-sided adhesive material 3, and the single-sided adhesive material 3 seals the bottom plate 2. In the present embodiment, the single-sided adhesive material 3 is provided with a clearance area at a corresponding position of the cleaning liquid storage part.

[0055] Embodiment 9

[0056] refer to Figure 1 On the basis of the first embodiment, a product label is provided on the surface of the top plate 1 of the ninth embodiment, and the product label is provided with relevant introduction of the chip, such as the chip name, the detection target of the chip, etc.

[0057] Furthermore, a QR code label is provided on the surface of the top plate 1 or the bottom plate 2, and a camera on an external detection instrument can scan the QR code on the QR code label to read corresponding data, such as the name of the chip, the detection target of the chip, product calibration information, etc. It should be noted that the setting position of the QR code label depends on the orientation position of the camera of the external detection instrument.

[0058] Embodiment 10

[0059] refer to Figure 1 On the basis of the first embodiment, the top plate 1 of the tenth embodiment is provided with a magnetic absorption hole 15 on the corresponding connecting tracks with the magnetic particle coating part 22, the cleaning area 23 and the detection area 24. The external magnet moves along the set direction of the magnetic absorption hole 15, driving the magnetic particles to move along the magnetic particle coating part 22, the cleaning area 23 and the detection area 24 in sequence. In addition, the magnetic absorption hole 15 is provided, so that the magnet can be closer to the bottom plate 2, avoiding the top plate 1 between the bottom plate 2 and the magnet, and greatly improving the reliability of magnetic absorption.

[0060] Embodiment 11

[0061] refer to Figure 1 On the basis of the first embodiment, the bottom plate 2 of the eleventh embodiment is provided with a waste liquid pool 27 interconnected with the cleaning area 23. The waste liquid pool 27 can collect the waste liquid after cleaning and reaction, which can reduce the interference of the waste liquid on the final detection and effectively improve the detection accuracy. There can be multiple waste liquid pools 27.

[0062] Embodiment 12

[0063] refer to Figure 1 On the basis of the first to eleventh embodiments, the base plate 2 of the twelfth embodiment further includes a luminescent liquid storage portion 26 interconnected with the detection area 24 , and the luminescent liquid storage portion 26 is provided with luminescent liquid inside.

[0064] After the sample is cleaned in the cleaning area 23, it enters the detection area 24. At this time, the luminescent liquid in the luminescent liquid storage part 26 is released, and the luminescent liquid reacts with the sample to emit a luminescent signal. The external detection instrument detects the intensity of the luminescent signal.

[0065] The luminescent liquid is pre-stored in the luminescent liquid storage unit 26, and the luminescent liquid is used to further wash the magnetic beads or enhance the luminescent signal. The luminescent liquid includes a substrate liquid and a luminescent enhancement liquid, the substrate liquid can be an acidic solution containing luminol or an acidic solution containing adamantane, and the luminescent enhancement liquid can be an alkaline solution containing a benzene derivative.

[0066] It is worth mentioning that, considering that the substrate liquid and the luminescence enhancement liquid should not be mixed and stored for a long time, the luminescence liquid storage part 26 can be provided with a first luminescence liquid storage part and a second luminescence liquid storage part, the first luminescence liquid storage part stores the substrate liquid, the second luminescence liquid storage part stores the luminescence enhancement liquid, and a luminescence liquid mixing area is provided on the bottom plate 2, and the luminescence liquid mixing area is connected to the detection area 24, the first luminescence liquid storage part and the second luminescence liquid storage part respectively. When the first luminescence liquid storage part and the second luminescence liquid storage part are released, the substrate liquid and the luminescence enhancement liquid enter the luminescence liquid mixing area and mix with each other, and then enter the detection area 24 after being evenly mixed.

[0067] The luminescent liquid storage part 26 is a sealed cavity, and the sealing material used is an elastic material or a high barrier film, specifically glass, plastic, rubber, aluminum foil or a high barrier film, wherein the sealing material can be composed of the same material or a combination of multiple materials. Under physical compression, the luminescent liquid storage part 26 can be partially ruptured, thereby releasing the stored luminescent liquid.

[0068] The labeled ligand is pre-stored in the labeled ligand storage unit 12. When the labeled ligand includes an enzyme-labeled ligand, the enzyme may be one or more of horseradish peroxide and alkaline phosphatase, and the ligand may be one or more of antigen, antibody, hapten and nucleic acid. The magnetic particle ligand solution is pre-stored in the magnetic particle coating unit 22. The magnetic particle ligand solution includes magnetic particles, sugars, buffer reagents, proteins, surfactants and preservatives. The magnetic particles include but are not limited to ferric oxide and ferric oxide compounds.

[0069] When the labeled ligand includes an enzyme-labeled ligand, the enzyme binds to or competes with the analyte in the sample to form an enzyme-labeled ligand; the magnetic particle label binds to or competes with the analyte in the sample to form a magnetic bead-labeled ligand, and the two ligands may be the same or different; the magnetically labeled ligand and the enzyme-labeled ligand include nucleic acids, antigens, monoclonal antibodies, polyclonal antibodies and hormone receptors, and the analytes in the sample include DNA, small molecules (drugs or drugs), antigens, antibodies, hormones, antibiotics, bacteria or viruses and other biochemical markers.

[0070] In this embodiment, the labeled ligand can be combined with the magnetic particle ligand solution (such as the double antibody sandwich method), or the labeled ligand can compete with the labeled ligand (such as the competitive method). The enzyme-labeled ligand can be the same as the magnetic particle ligand solution or different. Preferably, in one embodiment of the present invention, two different antibodies are selected as the labeled ligand and the magnetic particle ligand solution to detect the analyte by the double antibody sandwich method. In another embodiment of the present invention, an antigen and an antibody are selected as the labeled ligand and the magnetic particle ligand solution to detect the analyte by the competitive method.

[0071] Embodiment 13

[0072] refer to Figure 1 On the basis of the twelfth embodiment, the top plate 1 of the thirteenth embodiment is provided with a cleaning liquid clearance hole and a luminescent liquid clearance hole at corresponding positions of the cleaning liquid storage part 25 and the luminescent liquid storage part 26 on the bottom plate 2.

[0073] Since the cleaning liquid storage part 25 and the luminescent liquid storage part 26 are usually provided with a certain shape, such as a cylinder, etc., in order to adapt to the shapes of the cleaning liquid storage part 25 and the luminescent liquid storage part 26 and avoid local protrusion of the chip, a cleaning liquid clearance hole and a luminescent liquid clearance hole are set on the top plate 1.

[0074] In this embodiment, the cleaning liquid evacuation hole and the luminous liquid evacuation hole together constitute the evacuation hole 16 .

[0075] Embodiment 14

[0076] refer to Figure 1 On the basis of the first to eleventh embodiments, the labeled ligand storage unit 12 of the fourteenth embodiment is provided with a fluorescent agent. That is to say, the labeled ligand includes a ligand labeled with a fluorescent agent, and the fluorescent agent can be selected from one or more of acridinium ester, ABEI, fluorescent dye, fluorescent protein and fluorescent microspheres, and the ligand can be selected from one or more of antigen, antibody, hapten and nucleic acid. Among them, acridinium ester and ABEI can directly emit light after reacting with the luminescent liquid; while fluorescent dye, fluorescent protein and fluorescent microsphere need an excitation light source, but no luminescent liquid is required.

[0077] When the labeled ligand includes a ligand labeled with a fluorescent agent, the fluorescent agent binds to or competes with the analyte in the sample to form a fluorescent agent-labeled ligand; the magnetic particle ligand solution binds to or competes with the analyte in the sample to form a magnetic bead-labeled ligand, and the two ligands may be the same or different; the magnetic particle ligand solution and the ligand labeled with a fluorescent agent include nucleic acids, antigens, monoclonal antibodies, polyclonal antibodies and hormone receptors, and the analytes in the sample include DNA, small molecules (drugs or drugs), antigens, antibodies, hormones, antibiotics, bacteria or viruses and other biochemical markers.

[0078] In this embodiment, the labeled ligand can be combined with the magnetic particle ligand solution (such as double antibody sandwich method), or the labeled ligand can compete with the magnetic particle ligand solution (such as competition method). The ligand labeled with the fluorescent agent can be the same as the magnetic particle ligand solution or different. Preferably, in one embodiment of the present invention, two different antibodies are selected as the labeled ligand and the magnetic particle ligand solution to detect the analyte by the double antibody sandwich method.

[0079] Embodiment 15

[0080] This embodiment 15 provides a magnetic particle luminescent double-layer microfluidic detection system, the detection system includes: a magnetic particle luminescent double-layer microfluidic chip as described in embodiments 1 to 14; a magnet unit for driving the magnetic particles in the magnetic particle ligand solution to move; an extrusion unit for squeezing the labeled ligand storage part 12 and the cleaning liquid storage part 25 to allow the labeled ligand and the cleaning liquid to flow out; and a detection unit for detecting samples in the detection area 24.

[0081] The magnet unit includes a magnet and a driving part for driving the magnet to move. The driving part can be a linear motor, and the output shaft of the linear motor is fixedly connected to the magnet. After the linear motor is started, its output shaft extends to drive the magnet to move, and the magnet absorbs magnetic particles and drives the magnetic particles to move.

[0082] The squeezing unit may be a linear motor. After the linear motor is started, its output shaft extends to squeeze the labeled ligand storage part 12 and the cleaning liquid storage part 25, so that the labeled ligand and the cleaning liquid flow out respectively. Of course, a squeezing part may be fixedly mounted on the output shaft of the linear motor, and the squeezing part matches the shape and size of each storage part. The linear motor may be provided with one or more squeezing parts, and the squeezing part is driven to move by the output shaft of the linear motor, and the squeezing part then squeezes the labeled ligand storage part 12 and the cleaning liquid storage part 25, so that the labeled ligand and the cleaning liquid flow out respectively.

[0083] Among them, the detection unit can be selected as a photodiode, a photomultiplier tube or an avalanche photodiode. The sample enters the detection area after the mixing reaction through the above process. The mixed sample will have a luminescent signal. The detection unit collects the luminescent signal and obtains the detection result of the sample based on the strength of the luminescent signal.

[0084] When the magnetic particle luminescent double-layer microfluidic chip is provided with an air pump 14 , the squeezing unit can also be used to squeeze the air pump 14 to drive the liquid on the top plate 1 to flow.

[0085] When the magnetic particle luminescent double-layer microfluidic chip is provided with a luminescent liquid storage part 26 , the squeezing unit can also be used to squeeze the luminescent liquid storage part 26 to make the luminescent liquid flow out.

[0086] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A magnetic particle luminescent double-layer microfluidic chip, characterized in that: The chip comprises: A top plate, the top plate comprising a sample adding part, a labeled ligand storage part and a sample mixing area, wherein the labeled ligand storage part is provided with a labeled ligand, and the sample mixing area is communicated with the sample adding part and the labeled ligand storage part respectively; A bottom plate arranged on the top plate, the bottom plate comprising a flow guide area interconnected with the sample mixing area, a magnetic particle coating part interconnected with the flow guide area, a washing area interconnected with the magnetic particle coating part, a detection area interconnected with the washing area, and a washing liquid storage part interconnected with the washing area, wherein the flow guide area is provided with a groove whose height is lower than the bottom wall of the magnetic particle coating part, a flow guide part arranged on the groove and connected to the magnetic particle coating part, a truncation groove arranged below the front end of the flow guide part, and a blocking part arranged on the flow guide part, a magnetic particle ligand solution is arranged inside the magnetic particle coating part, and a washing liquid is arranged inside the washing liquid storage part; The sample adding part comprises a sample adding port and a cover for opening or closing the sample adding port, and the sample adding part further comprises a rubber ring arranged on the sample adding port.

2. The magnetic particle luminescent double-layer microfluidic chip according to claim 1, characterized in that: The top plate also includes an air pump interconnected with the sample adding part.

3. The magnetic particle luminescent double-layer microfluidic chip according to claim 2, characterized in that: The top plate is provided with elastic members at positions corresponding to the air pump and the sample mixing area.

4. The magnetic particle luminescent double-layer microfluidic chip according to claim 2, characterized in that: A porous elastic member is arranged inside the air pump.

5. The magnetic particle luminescent double-layer microfluidic chip according to claim 1, characterized in that: The top plate and the bottom plate are both provided with limiting notches at positions corresponding to each other.

6. The magnetic particle luminescent double-layer microfluidic chip according to claim 1, characterized in that: The top plate is provided with a first buckle or a first slot, and the bottom plate is provided with a second slot or a second buckle, and the top plate and the bottom plate are buckled with each other through the mutual cooperation between the first buckle and the second slot, or through the mutual cooperation between the first slot and the second buckle.

7. The magnetic particle luminescent double-layer microfluidic chip according to claim 1, characterized in that: A single-sided adhesive material is provided on a portion or the entire area of ​​at least one side of the bottom plate.

8. The magnetic particle luminescent double-layer microfluidic chip according to claim 1, characterized in that: A product label is provided on the surface of the top plate or the bottom plate, and a QR code label is provided on the surface of the top plate or the bottom plate.

9. The magnetic particle luminescent double-layer microfluidic chip according to claim 1, characterized in that: The top plate is provided with magnetic absorption holes on the corresponding connecting tracks with the magnetic particle coating part, the cleaning area and the detection area.

10. The magnetic particle luminescent double-layer microfluidic chip according to claim 1, characterized in that: The bottom plate also includes a waste liquid pool interconnected with the cleaning area.

11. The magnetic particle luminescent double-layer microfluidic chip according to any one of claims 1 to 10, characterized in that: The bottom plate further includes a luminescent liquid storage portion interconnected with the detection area, and luminescent liquid is arranged inside the luminescent liquid storage portion.

12. The magnetic particle luminescent double-layer microfluidic chip according to claim 11, characterized in that: The top plate is provided with a cleaning liquid evacuation hole and a luminescent liquid evacuation hole at corresponding positions of the cleaning liquid storage part and the luminescent liquid storage part.

13. The magnetic particle luminescent double-layer microfluidic chip according to any one of claims 1 to 10, characterized in that: The labeled ligand storage part has fluorescent liquid inside.

14. A magnetic particle luminescent double-layer microfluidic detection system, characterized in that: The detection system comprises: The magnetic particle luminescent double-layer microfluidic chip according to any one of claims 1 to 13; A magnet unit for driving the magnetic particles in the magnetic particle ligand solution to move; An extrusion unit for squeezing the labeled ligand storage part and the cleaning solution storage part to allow the labeled ligand and the cleaning solution to flow out; A detection unit for detecting the luminescent signal in the detection zone.

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