A fully automatic magnetic particle chemiluminescence detection disc and detection device

Abstract

The present invention provides a fully automatic magnetic microparticle chemiluminescence detection disc and detection device, belonging to the field of medical inspection, including: a sample processing module; an incubation module, the incubation module is located on the distal end side compared to the sample processing module, the incubation module includes an incubation pool, a quantitative channel and an antibody storage cavity connected to the incubation pool through a first valve, the antibody storage cavity is provided with magnetic beads coated with antibodies, the sample processing module is connected to the proximal side of the first channel through a second valve and is located in the upper area corresponding to the incubation pool, the distance between the proximal end of the antibody storage cavity and the rotation center is less than the distance between the incubation pool and the distal end of the quantitative channel and the rotation center; a cleaning reaction module, the cleaning reaction module is located downstream of the incubation module and is adapted to the incubation module. The present invention adopts non-fixed magnetic beads coated with antibodies, which increases the degree of reaction between antibodies and samples, and has a better cleaning effect.

Description

Technical Field

[0001] The present application relates to the field of medical testing, and in particular to a fully automatic magnetic particle chemiluminescence detection disc and a detection device. Background Art

[0002] In recent years, the field of bioanalysis technology has developed rapidly, and many important research directions have emerged. Microfluidic chip analysis technology is the most active one among them, and has gained wide attention in both scientific research and application fields. As a new type of analysis and detection platform, microfluidic chips have the advantages of high throughput, integration, portability, easy operation, and low cost, and have been widely used in many fields.

[0003] The current chemiluminescence detection is suitable for samples such as serum. During the detection process, the sample needs to be centrifuged to obtain serum. The process is complicated, cumbersome and time-consuming, and lacks sufficient elution to eliminate the interference caused by nonspecific adsorption. Secondly, in the prior art, since the magnetic beads coated with antibodies are fixed, the degree of reaction between the antibodies on the magnetic beads and the samples is low and the reaction is insufficient. Secondly, the fixed magnetic beads lead to insufficient subsequent washing, which makes the detection results unstable. Summary of the invention

[0004] The purpose of the present invention is to provide a fully automatic magnetic microparticle chemiluminescence detection disc and detection device. The use of non-fixed antibody-coated magnetic beads increases the degree of reaction between the antibody and the sample, and has a better cleaning effect, making the subsequent detection results more accurate.

[0005] The present invention adopts the following technical solution:

[0006] A fully automatic magnetic particle chemiluminescence detection disk, comprising: a sample processing module; an incubation module, wherein the incubation module is located on the distal end side compared to the sample processing module, the incubation module comprises an incubation pool, a quantitative channel and an antibody storage cavity connected to the incubation pool via a first valve, magnetic beads coated with antibodies are arranged in the antibody storage cavity, the sample processing module is connected to the proximal side of the first channel via a second valve and is located in the upper area corresponding to the incubation pool, the distance between the proximal end of the antibody storage cavity and the rotation center is smaller than the distance between the incubation pool and the distal end of the quantitative channel and the rotation center; a cleaning reaction module, wherein the cleaning reaction module is located downstream of the incubation module and is adapted to the incubation module.

[0007] Furthermore, the quantitative channel and the antibody storage cavity are respectively located on both sides of the incubation pool, the bottom of the incubation pool and the bottom of the antibody storage cavity are located on the same arc surface, and the first valve is located on the arc surface.

[0008] Furthermore, the cleaning reaction module includes at least one cleaning pool and at least one reaction pool; the proximal sides of the cleaning pool, reaction pool and incubation pool are connected through a first channel; a reagent storage module is connected to the proximal side of the first channel through a third valve and is arranged corresponding to the upper area of ​​the cleaning pool, and a substrate storage chamber is connected to the proximal side of the first channel through a fourth valve and is arranged corresponding to the upper area of ​​the reaction pool.

[0009] Furthermore, a sealing liquid storage chamber is connected to the proximal side of the first channel via a fifth valve.

[0010] Further, the reagent storage module includes a corresponding number of reagent storage chambers as the cleaning pool, the reagent storage chamber is connected to the proximal side of the first channel through a third valve and is located in the area above the corresponding cleaning pool; or the reagent storage module includes a reagent storage chamber and a corresponding number of quantitative distribution chambers as the cleaning pool, the quantitative distribution chamber is connected to the proximal side of the first channel through a third valve and is located in the area above the corresponding cleaning pool, the proximal end of the quantitative distribution chamber is connected in series through a distribution channel, one end of the distribution channel is connected to the proximal end of the first overflow chamber, and the other end is connected to the distal end of the reagent storage chamber through a sixth valve.

[0011] Furthermore, the depth of the first channel is smaller than that of the waste liquid tank, the incubation tank, the cleaning tank and the reaction tank.

[0012] Furthermore, the sample processing module includes: a sample chamber, the sample chamber having a sample addition port; a buffer storage module; a sample centrifugal quantitative module, the distal end of the sample chamber is connected with the proximal end of the sample centrifugal quantitative module and the overflow chamber through a seventh valve; a mixing chamber, the proximal end of the mixing chamber is connected with the buffer storage module through an eighth valve and is connected with the sample centrifugal quantitative module through a ninth valve; the distal end of the mixing chamber is connected to the proximal side of the first channel of the incubation module of the same number as the second valve through at least one second valve and the connection position is located in the upper area corresponding to the incubation pool.

[0013] Furthermore, the quantitative channel is connected to a waste liquid pool or a parallel incubation module; and a corresponding cleaning reaction module is provided downstream of the incubation module.

[0014] Further, the buffer storage module is a buffer storage cavity; or the buffer storage module includes a buffer storage cavity and a buffer quantitative pool, the proximal end of the buffer quantitative pool is connected to the buffer storage cavity through a tenth valve, the distal end of the buffer quantitative pool is connected to the mixing cavity through an eighth valve, and the proximal end of the buffer quantitative pool is provided with an overflow channel, and the overflow channel is connected to the overflow cavity.

[0015] Furthermore, the sample centrifugal quantitative module includes a serum quantitative pool, a sedimentation pool and a check valve connecting the quantitative pool and the sedimentation pool, which are arranged in sequence away from the rotation center; one end of the ninth valve is connected to the distal end of the serum quantitative pool.

[0016] A detection device adopts the above-mentioned fully automatic magnetic particle chemiluminescence detection disk for detection.

[0017] The present invention provides a highly integrated microfluidic chip, which has the following beneficial technical effects compared with the prior art:

[0018] (1) The present invention sets the antibody-coated magnetic beads in the antibody storage chamber, opens the first valve at the right time, and then the processed sample is fully mixed with the antibody-coated magnetic beads, thereby improving the binding degree of the antigen and antibody in the sample. In order to achieve the effect of accurate quantification, the present application defines that the distance between the proximal end of the antibody storage chamber and the rotation center is less than the distance between the incubation pool and the distal end of the quantitative channel and the rotation center. Under the condition of the rotation of the fully automatic magnetic microparticle chemiluminescence detection disk, when the second valve is opened, the sample in the sample processing module continues to fill the incubation pool under the action of centrifugal force. When the incubation pool is completely filled, the excess processed sample flows out through the quantitative channel to quantify the sample, thereby improving the accuracy and stability of the test result. Secondly, when the sample in the sample processing module is completely released, the first valve is opened, and the sample in the incubation pool enters the antibody storage chamber and fully contacts the antibody-coated magnetic beads. Since part of the processed sample enters the antibody storage chamber, the liquid level in the incubation pool drops, thereby avoiding the speed fluctuation during the rotation of the detection disk, which causes part of the liquid to flow out through the quantitative channel. Thirdly, the distance between the antibody storage cavity and the rotation center is set to be small so that the sample processing module will not overflow into the antibody storage cavity when filling the incubation pool, or will not enter the downstream cleaning reaction module through the wall of the antibody storage cavity close to the rotation center.

[0019] (2) The present invention creatively proposes to arrange the incubation pool, the cleaning pool and the reaction pool in sequence along the circumferential direction, and at the same time, by setting a first channel at the proximal end of each chamber, and sealing and filling the first channel, the quantitative channel and the cavity of the distal end chamber with mineral oil, the possibility of cross-contamination between liquid reagents is completely avoided; in addition, through the first channel set at the proximal end of each chamber, the magnetic beads can be smoothly transferred in each chamber only by centrifugal rotation. In addition, since the sealing liquid has a repulsive effect on the carried aqueous phase substances, it can reduce the magnetic beads in the process of circulation. The reagent or impurities in the upstream cavity is brought into the downstream cavity, which improves the cleaning and impurity removal effect of the magnetic beads, and further improves the accuracy and stability of the detection results. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0021] Figure 1 A schematic diagram of the structure of a fully automatic magnetic microparticle chemiluminescence detection disk provided in an embodiment of the present invention;

[0022] Figure 2 A schematic diagram of the structure of an incubation module and a cleaning reaction module provided in an embodiment of the present invention;

[0023] Figure 3 A schematic diagram of another reagent storage module structure provided by an embodiment of the present invention;

[0024] Figure 4 A schematic diagram of the structure of a sample processing module provided in an embodiment of the present invention;

[0025] Figure 5 A schematic diagram of another sample processing module structure provided by an embodiment of the present invention;

[0026] Figure 6 A schematic diagram of the structure of another incubation module and a cleaning reaction module provided in an embodiment of the present invention;

[0027] Figure 7 A schematic diagram of the structure of another buffer storage module provided by an embodiment of the present invention;

[0028] Figure 8 A schematic diagram of the structure of a sample centrifugal quantitative module provided in an embodiment of the present invention;

[0029] Fig. 9 A schematic diagram of the check valve structure provided in an embodiment of the present invention.

[0030] Figure numerals: 001, first gas circuit; 002, second gas circuit; 003, third gas circuit; 004, fourth gas circuit; 01, first valve; 02, second valve; 03, third valve; 04, fourth valve; 05, fifth valve; 06, sixth valve; 07, seventh valve; 071, first flow channel; 072, second flow channel; 08, eighth valve; 09, ninth valve; 010, tenth valve; 1, sample processing module; 10, sample chamber; 100, sample injection port; 11, buffer storage module; 110, buffer storage chamber; 111, buffer quantitative pool; 112, overflow channel; 12, sample centrifugal quantitative module; 120, serum quantitative pool ; 121. sedimentation tank; 122. check valve; 1220. valve chamber; 1221. connecting part; 1222. deformation part; 1223. counterweight part; 13. mixing chamber; 2. incubation module; 20. incubation tank; 21. quantitative channel; 22. antibody storage chamber; 23. waste liquid tank; 3. cleaning reaction module; 30. cleaning tank; 31. reaction tank; 32. first channel; 33. substrate storage chamber; 34. sealing liquid storage chamber; 4. reagent storage module; 40. reagent storage chamber; 41. quantitative distribution chamber; 42. distribution channel; 50. first overflow chamber; 51. second overflow chamber; 511. first cavity; 512. second cavity. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Generally, the components of the embodiments of the present invention described and shown in the drawings here can be arranged and designed in various different configurations.

[0032] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. 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.

[0033] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, further definition and explanation thereof is not required in subsequent drawings.

[0034] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inside", "outside", etc. indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings, or the positions or positional relationships in which the product of the invention is usually placed when in use. They are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific position, be constructed and operated in a specific position, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", "third", etc. are only used to distinguish the description, and cannot be understood as indicating or implying relative importance.

[0035] In the description of the present invention, it is also necessary to explain that, unless otherwise clearly specified and limited, the terms "set", "install", "connect", and "connect" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be a connection between two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0036] In the present invention, the "distal end" and "proximal end" are obtained by comparing the distances relative to the rotation center O. For example, the side of a chamber close to the rotation center is the proximal end, and the side away from the rotation center is the distal end.

[0037] The "sealing liquid" is a hydrophobic liquid with a density lower than that of the liquid in the first working chamber, and is inert and will not react with substances in the system, such as mineral oil, etc. The specific composition of the sealing liquid is not limited here.

[0038] The embodiment of the present invention discloses a fully automatic magnetic microparticle chemiluminescence detection disk, such as Figure 1 As shown, it includes: a sample processing module 1 (such as Figure 1 Incubation module 2, the incubation module 2 is located at the distal end side compared to the sample processing module 1, the incubation module 2 includes an incubation pool 20, a quantitative channel 21 and an antibody storage chamber 22 connected to the incubation pool 20 through a first valve 01, the antibody storage chamber 22 is provided with magnetic beads coated with antibodies (not shown in the figure), the sample processing module 1 is connected to the proximal side of the first channel 32 through a second valve 02 and is located in the upper area corresponding to the incubation pool 20, the distance L1 between the proximal end of the antibody storage chamber 22 and the rotation center O is smaller than the distances L2 and L3 between the distal end of the incubation pool 20 and the quantitative channel 21 and the rotation center O; a cleaning reaction module 3 (as shown by the fan-shaped dotted line in the figure), the cleaning reaction module 3 is located downstream of the incubation module 2 and is adapted to the incubation module 2.

[0039] The present invention arranges the antibody-coated magnetic beads in the antibody storage chamber 22 and opens the first valve 01 at an appropriate time, so that the processed sample is fully mixed with the antibody-coated magnetic beads, thereby improving the degree of antigen-antibody binding in the sample. In order to achieve the effect of precise quantification, the present application stipulates that the distance L1 between the proximal end of the antibody storage chamber 22 and the rotation center O is smaller than the distance L2 and L3 between the distal end of the incubation pool 20 and the quantitative channel 21 and the rotation center O. Under the condition of rotation of the fully automatic magnetic particle chemiluminescence detection disk, when the second valve 02 is opened, the sample in the sample processing module 1 continues to fill the incubation pool 20 under the action of centrifugal force. When the incubation pool 20 is completely filled, the excess processed sample flows out through the quantitative channel 21 to quantify the sample, thereby improving the accuracy and stability of the detection result. Secondly, when the sample in the sample processing module 1 is completely released, the first valve 01 is opened, and the sample in the incubation pool enters the antibody storage cavity 22, and is fully contacted with the magnetic beads coated with antibodies. Since part of the processed sample enters the antibody storage cavity 22, the liquid level in the incubation pool drops, so as to avoid the speed fluctuation during the rotation of the detection disk, which causes part of the liquid to flow out through the quantitative channel 21. Thirdly, the distance between the antibody storage cavity and the rotation center O is set to be small, so that when the sample processing module 1 fills the incubation pool 20, it will not overflow into the antibody storage cavity 22 (at this time, the antibody storage cavity is a cavity with an opening toward the rotation center), or will not enter the downstream cleaning reaction module 3 through the wall of the antibody storage cavity 22 close to the rotation center O (at this time, the antibody storage cavity is a closed cavity, and the antibody storage cavity communicates with the outside world through the opening and closing of the first valve 01).

[0040] It should be noted that the "upper region" in the present invention refers to the sector formed by the lines connecting the two ends of the cavity opening and the rotation center, and the area within this sector can be called the upper region. Figure 1 The lines between the two ends of the opening of the incubation pool and the rotation center form a sector S, and the connection position P0 of the second valve 02 and the first channel is located in the sector. The reason for this limitation is to ensure that the processed samples in the sample processing module 1 can smoothly enter the incubation pool under the action of centrifugal force.

[0041] Preferably, in order to prevent the processed sample from overflowing from the incubation tank 20, the length of L1 is at most 0.95 times the length of L2 or L3 and the difference between L1 and L2 or L3 is not less than 0.5 mm.

[0042] Secondly, the quantitative channel 21 of the present application can be used to connect to the waste liquid pool 23, such as Figure 1 as shown, or connect another incubation module (this solution will be described later).

[0043] Specifically, Figure 1 , the quantitative channel 21 and the antibody storage chamber 22 are respectively located on both sides of the incubation pool 20, the bottom of the incubation pool 20 and the bottom of the antibody storage chamber 22 are located on the same arc surface, and the first valve 01 is located on the arc surface. The first valve is set at the bottom to avoid the formation of a dead zone between the incubation pool and the antibody storage chamber during the mixing process, resulting in insufficient reaction between the treated sample and the antibody-coated magnetic beads; secondly, the present invention attracts magnetic beads through an external magnet, and under the movement of the external magnet, the magnetic beads are driven to move in the incubation pool and the downstream cleaning reaction module 3. The first valve 01 is set at the bottom to facilitate the magnet to bring as many magnetic beads as possible into the downstream chamber, avoiding some magnetic beads from falling due to the cavity structure.

[0044] Specifically, Figure 2 , the cleaning reaction module 3 includes at least one cleaning pool 30 and at least one reaction pool 31. In the embodiment of the present invention, two cleaning pools are provided, which can be distinguished by upstream cleaning pools and downstream cleaning pools; the proximal sides of the cleaning pool 30, the reaction pool 31 and the incubation pool 20 are connected through the first channel 32; a reagent storage module 4 is connected to the proximal side of the first channel 32 through the third valve 03 and is provided in the upper area of ​​the cleaning pool 30, and a substrate storage chamber 33 is connected to the proximal side of the first channel 32 through the fourth valve 04 and is provided in the upper area of ​​the reaction pool 31. In this embodiment, the reagent storage module 4 is two reagent storage chambers, the number of which is the same as that of the cleaning pools, and the reagent storage chamber is connected to the proximal side of the first channel 32 through the third valve 03 and is located in the upper area of ​​the corresponding cleaning pool 30. In this embodiment, the reagent storage chamber is provided with reagents and quantified, and the quantified volume must be less than the maximum volume that the corresponding cleaning pool 30 can accommodate.

[0045] Preferably, the first channel 32 is in the shape of an arc or a spiral line, and the incubation pool, cleaning pool and reaction pool are located on one side of the distal end of the first channel and are arranged in sequence along the direction of the spiral line, or the incubation pool, cleaning pool and reaction pool are arranged in sequence along the arc direction.

[0046] Specifically, Figure 2, and the sealing liquid storage chamber 34 is connected to the proximal side of the first channel 32 through the fifth valve 05. The sealing liquid storage chamber 34 stores sealing liquid, which is used to fill the cavities of the first channel 32, the quantitative channel 21 and the distal end chamber thereof. Through the sealing of the sealing liquid, the cross-contamination problem between different functional chambers is completely avoided; secondly, by setting the first channel at the proximal end of each chamber, the magnetic beads can be smoothly transferred in each chamber while achieving sealing; finally, since the sealing liquid has an incompatible effect with the water phase, when the magnetic beads with the complex enter the downstream chamber from the upstream chamber, the sealing liquid has the effect of repelling the carried water phase substances, so as to reduce the magnetic beads in the process of circulation, and bring the reagents or impurities in the upstream chamber into the downstream chamber, thereby improving the cleaning and impurity removal effect of the magnetic beads. In order to facilitate the reagents and substrates to enter the corresponding chambers and the sealing liquid to fill the cavities well, the first channel 32 is provided with a first gas path 001 on the proximal side, and the first gas path 001 is connected to the outside to avoid the high pressure in the chamber causing the liquid to flow unsmoothly.

[0047] like Figure 2 , the depth of the first channel 32 (the distance perpendicular to the paper surface) is less than the depth of the incubation tank, the cleaning tank and the reaction tank. The preferred depth of the first channel is 1 / 5-1 / 3 of the depth of the incubation tank, the cleaning tank or the reaction tank, and the depth of the first channel is not less than 100µm and not more than 2mm. The reason for limiting the depth of the first channel is that the problem of possible cross-contamination of the sealing liquid during the filling process is taken into account. Specifically, when the processed sample has filled the incubation tank, the reagent storage module 4 has filled the cleaning tank, and the substrate storage chamber has filled the reaction tank, the fifth valve is opened and the sealing liquid fills the downstream cavity, such as Figure 1 The sealing liquid first fills the upper cavity of the incubation tank, and then fills the upper cavity of the cleaning tank and the reaction tank. The sealing liquid needs to flow through different chambers. If the thickness of the first channel is thick, the sealing liquid flow rate is large, which disturbs the already filled samples, reagents or substrates, and carries the substances in the upstream chamber into the downstream, causing cross contamination. Therefore, the present invention limits the depth of the first channel, and the depth of the first channel has a certain correlation with the depth of each chamber, which reduces the disturbance of the sealing liquid to the liquid in each chamber while ensuring the filling efficiency.

[0048] As a preferred embodiment, Figure 3, the reagent storage module 4 includes a reagent storage chamber 40 and a quantitative distribution chamber 41 corresponding to the number of the cleaning pools 30, the quantitative distribution chamber 41 is connected to the proximal side of the first channel 32 through the third valve 03 and is located in the upper area of ​​the corresponding cleaning pool 30, the proximal end of the quantitative distribution chamber 41 is connected in series through the distribution channel 42, one end of the distribution channel 42 is connected to the proximal end of the first overflow chamber 50, and the other end is connected to the distal end of the reagent storage chamber 40 through the sixth valve 06. The above structure can pre-add the corresponding reagent to the reagent storage chamber 40 without the need for precise quantification, reducing the operating accuracy requirements of the operator and facilitating subsequent mass production. The distribution channel 42 is in the shape of an arc or a gradually receding spiral, preferably a gradually receding spiral, so that the reagent storage chamber 40 can fill the quantitative distribution chamber 41 in sequence; the quantitative distribution chamber 41 is in the shape of a fan ring, and is connected to the distribution channel on the side close to the rotation center. Since the fan ring opening is in a contracted shape, the accuracy of the reagent quantification is increased. In order to further ensure the quantitative accuracy of the quantitative distribution chamber, the depth of the distribution channel (i.e., the distance perpendicular to the paper surface) is 1 / 3-1 / 2 of the depth of the quantitative distribution chamber 41, and the depth of the distribution channel is not less than 100µm and not more than 5mm, so as to solve the problem of inaccurate filling amount of reagent due to the high degree of turbulence of the reagent flow during the filling process.

[0049] Specifically, Figure 4 , 5 As shown in Figures 7 and 8, the sample processing module 1 includes: a sample chamber 10, the sample chamber having a sample injection port 100; a buffer storage module 11; a sample centrifugal quantitative module 12, the distal end of the sample chamber 10 is connected to the proximal end of the sample centrifugal quantitative module 12 and the second overflow chamber 51 through the seventh valve 07; a mixing chamber 13, the proximal end of the mixing chamber 13 is connected to the buffer storage module 11 through the eighth valve 08 and is connected to the sample centrifugal quantitative module 12 through the ninth valve 09; the distal end of the mixing chamber 13 is connected to the proximal side of the first channel 32 of the incubation module of the same number as the second valve 02 through at least one second valve 02, and the connection position is located in the upper area corresponding to the incubation pool 20. Figure 4 In the embodiment, the second valve 02 is provided with a Figure 5 As shown, two second valves 02 are provided, and three or more can also be provided based on actual needs. Different incubation modules use different or the same antibodies to detect protein antigens, use different antibodies to detect different antigens, and use the same antibody to detect the same processed sample to ensure the accuracy of the test results. Preferably, the sample chamber is provided with a third gas path 003 on one side close to the rotation center to facilitate the smooth addition of the sample, and the mixing chamber 13 has a fourth gas path 004 to facilitate the liquid to enter / flow out of the mixing pool and the pressure balance during the mixing process, so that the mixing effect is good and the liquid path reliability is high.

[0050] like Figure 4 and 5 The seventh valve 07 in the present invention includes a first flow channel 071 and a second flow channel 072. The first flow channel 071 is connected to the second flow channel 072. The first flow channel 071 extends along the radial direction of the fully-automatic magnetic particle chemiluminescence detection disk to connect the sample chamber and the sample centrifugation quantitative module. The second flow channel 072 extends along the circumference of the fully-automatic magnetic particle chemiluminescence detection disk and is connected to the second overflow chamber 51. The second overflow cavity 51 includes a first cavity 511 and a second cavity 512, the first cavity 511 and the second cavity 512 are fan-shaped, the proximal side of the first cavity 511 is connected to the distal side of the second cavity 512, the depth of the first cavity 511 is less than the depth of the second cavity 512, preferably, the depth of the second cavity 512 (i.e., the distance perpendicular to the paper surface) is 2-5 times the depth of the first cavity 511, and the depth of the second cavity 512 is not less than 2mm and not more than 10mm, the second flow channel 072 is connected to the second cavity 512, preferably, the depth of the second flow channel 072 (i.e., the distance perpendicular to the paper surface) is 1 / 5-1 / 3 of the depth of the second cavity 512, and the depth of the second flow channel 072 is not less than 0.5mm and not more than 2mm. Since the second overflow chamber 51 in the present application can not only contain liquid but also fix the liquid due to surface tension; in addition, the second flow channel 072 can also fix the quantified liquid. Through the above arrangement, it is avoided that the detection disk is in a low speed or static state, and the liquid fluctuation causes the backflow into the sample centrifugal quantification module 12 or the quantified liquid overflows into the second overflow chamber 51. Optionally, a second air path 002 is set on the proximal side of the second overflow chamber 51 for connecting the cavity with the outside.

[0051] Specifically, Figure 6 As shown, the quantitative channel 21 is connected to the parallel incubation module 2; the downstream of the incubation module 2 has a corresponding cleaning reaction module 3. When the processed sample is filled into the incubation pool 20 in the parallel incubation module 2 through the quantitative channel 21, the excess liquid flows into the waste liquid pool 23. The above arrangement increases the number of detection items, improves detection efficiency and accuracy.

[0052] Specifically, Figure 7 As shown, the buffer storage module 11 is a buffer storage chamber. The buffer storage chamber is communicated with the mixing chamber 13.

[0053] In another preferred embodiment, Figure 4Or as shown in Figure 5, the buffer storage module 11 includes a buffer storage chamber 110 and a buffer quantitative pool 111, the proximal end of the buffer quantitative pool 111 is connected to the buffer storage chamber 110 through the tenth valve 010, the distal end of the buffer quantitative pool 111 is connected to the mixing chamber 13 through the eighth valve 08, and the proximal end of the buffer quantitative pool 111 is provided with an overflow channel 112, and the overflow channel 112 is connected to the second overflow chamber 51.

[0054] Optionally, the depth of the overflow channel 112 is less than the depth of the buffer quantitative pool 111. Preferably, the depth of the overflow channel (i.e., the distance perpendicular to the paper surface) is 1 / 4-1 / 2 of the depth of the buffer quantitative pool 111, and the depth of the overflow channel is not less than 0.5 mm and not more than 2 mm. The overflow channel is fan-shaped, and the side of the overflow channel away from the rotation center is connected to the proximal side of the buffer quantitative pool 111.

[0055] like Figure 8 As shown, the sample centrifugal quantitative module 12 includes a serum quantitative pool 120, a sedimentation pool 121, and a check valve 122 connecting the quantitative pool and the sedimentation pool, which are sequentially arranged away from the rotation center O; one end of the ninth valve 09 is connected to the distal end of the serum quantitative pool 120. Under the action of centrifugal force, large particles in the sample enter the sedimentation pool 121, and the serum is kept in the serum quantitative pool 120 to achieve separation. A check valve 122 is arranged between the sedimentation pool 121 and the serum quantitative pool 120 to prevent large particles from entering the sedimentation pool 121 and then backflowing into the serum quantitative pool, causing serum contamination.

[0056] The serum quantitative pool adopts a funnel shape to facilitate large particles to enter the sedimentation tank through the check valve 122. The specific structure of the check valve 122 is as follows: Fig. 9 As shown, the check valve has a valve cavity 1220, a connecting portion 1221, a deformation portion 1222 and a counterweight portion 1223. The connecting portion 1221 is arranged on the inner wall of the valve cavity 1220 and extends in a funnel shape toward a side away from the rotation center O. The deformation portion 1222 is connected to the connecting portion 1221, and its core is closed. The end of the deformation portion is provided with a counterweight portion 1223, and the counterweight portion 1223 is located at 1 / 3-2 / 3 of the projection of the connecting portion 1221 and the deformation portion 1222 in the horizontal direction of the check valve, such as P1 is located at the midpoint of L4 in the figure. Preferably, as Fig. 9As shown, the connecting portion 1221, the deformation portion 1222 and the counterweight portion 1223 are all made of the same material, and the difference in deformation capacity is achieved by changing the thickness at different positions. In the present application, the counterweight portion 1223 is annular and the connecting portion 1221 is trumpet-shaped. Both are difficult to deform. Therefore, under the action of centrifugal force, the counterweight portion 1223 applies a pulling force to the deformation portion 1222, thereby opening the closed core of the deformation portion 1222; when the centrifugal force is cancelled, the core of the deformation portion 1222 is closed and the check valve 122 is closed. If the liquid in the sedimentation tank 121 fluctuates and pushes the counterweight portion 1223 to move upward, the counterweight portion 1223 squeezes the deformation portion, thereby sealing the core of the deformation portion 1222 more tightly. The check valve in the present application has a simple structure and is easy to manufacture and process; secondly, since the substances to be precipitated are cells and other substances of different sizes and have biological viscosity, the use of a brush-like structure facing away from the rotation center is likely to cause residual cell substances in the brush body, affecting the separation effect, while the check valve proposed in the present application has a smooth wall and no seams, so there are fewer residues such as cells.

[0057] It can be understood that the deformation part 1222 is elastic and can be pulled to enlarge or reduce the through hole in the core. The core seal can be a non-elastic seal or an elastic seal, that is, under the condition of no centrifugal force, the core seal of the deformation part 1222 only needs to be in a closed state.

[0058] The present invention is provided with a plurality of valves, such as the first valve, the second valve, the third valve, ... the tenth valve, etc. The form and combination of the valves are not limited, as long as they can be opened and closed in the required steps, and can be active valves or passive valves. Specifically, a manually operated mechanical valve or a phase change valve in the prior art can be used, or a capillary channel opened or closed under centrifugal action can be used. In order to reduce production costs while maintaining detection accuracy, it is preferred to use a capillary channel for control, specifically, a capillary tube, the size of which is from hundreds of microns to millimeters, and different diameters and lengths can be used to achieve the effect of opening or closing under different centrifugal forces. A phase change valve refers to a valve body that can produce a closing and opening effect when external conditions change.

[0059] In order to further increase the automation of the detection disk, such as realizing the opening or closing of various valve bodies in a preset order only by centrifugation, preferably, the third valve, the fourth valve, the fifth valve, the sixth valve, the seventh valve and the tenth valve are capillary channels, the second valve, the eighth valve and the ninth valve are siphon valves, and the first valve adopts a capillary channel or a phase change valve. The above-mentioned setting takes into account the special design for whole blood samples. Specifically, the connection of the mixing chamber 13 basically adopts a siphon valve, which completely prohibits the flow of unwanted liquid to a certain extent. For example, due to the siphon effect of the capillary channel, there is a possibility that liquid flows from the upstream cavity to the downstream cavity to a certain extent. The siphon valve strictly prohibits the flow of incompletely mixed samples from the mixing chamber 13 to the downstream chamber, and strictly prohibits the release of incompletely processed samples and unquantified buffer solutions into the mixing chamber 13, so as to improve the accuracy and stability of the detection; and the control accuracy of other valve bodies except the siphon valve can relatively meet the requirements of detection stability and accuracy, and no strict control is required, and the above structure is simple, and automation can be achieved without manual control of the opening and closing of the valve body, and the first valve can also achieve the corresponding function by using a capillary channel, but the use of a phase change valve can improve the protection of the coated antibody magnetic beads and improve the safety of storage.

[0060] The embodiment of the present invention further provides a detection device, which uses the above-mentioned fully automatic magnetic particle chemiluminescence detection disk for detection.

[0061] For convenience, Figure 1 , 2 The principle is explained using the fully automatic magnetic microparticle chemiluminescence detection disk in and 4 as an example.

[0062] AMaterial preparation:

[0063] (1) The buffer storage module is pre-stored with sample diluent for diluting the serum sample.

[0064] (2) Mineral oil is pre-stored in the sealing liquid storage chamber and is used to fill the first channel and the cavity in the immune response module to achieve isolation and sealing of each functional chamber in the incubation module and the cleaning reaction module.

[0065] (3) The upstream reagent storage chamber is pre-stored with cleaning solution 1, which is used for the first cleaning of the immune complex formed by the magnetic beads-antigen-antibody.

[0066] The downstream reagent storage chamber is pre-stored with a cleaning solution 2 for a second cleaning of the immune complex formed by the magnetic beads-antigen-antibody.

[0067] The substrate storage chamber is pre-stored with an enzymatic reaction substrate, which generates an enzymatic chemical reaction when mixed with the immune complex.

[0068] The antibody storage chamber is pre-stored with magnetic beads and secondary antibodies that participate in the reaction.

[0069] B reaction process is as follows:

[0070] 1. Add whole blood sample into the sample chamber through the sample port.

[0071] 2. Start the centrifuge to 500rpm, the whole blood sample enters the serum weighing tank and sedimentation tank through the connecting tube, the excess whole blood sample enters the second overflow chamber through the connecting tube, increase the centrifugal speed to 1000rpm, and continue centrifugation for 60s to separate blood cells from serum. Since the density of blood cells is greater than that of serum, blood cells enter the sedimentation tank. At the same time, due to the presence of the check valve, it can prevent the blood cells and serum from re-dissolving after the centrifugation stops.

[0072] 3. At the same time, the tenth valve is opened, the sample diluent in the buffer storage module enters the buffer quantitative pool, and the excess liquid enters the second overflow chamber.

[0073] 4. Reduce the centrifugal speed to 100 rpm. The separated serum sample will fill the siphon tube of the ninth valve, and the sample diluent will complete the filling of the siphon tube of the eighth valve.

[0074] 5. Increase the centrifugal speed to 800 rpm again, and the quantified serum sample and sample diluent enter the mixing tank to complete mixing.

[0075] 6. Reduce the centrifugal speed to 100 rpm again, and the diluted serum sample will fill the siphon pipe of the second valve.

[0076] 7. Increase the centrifugal speed to 800 rpm again, the diluted serum sample enters the incubation pool, and the excess sample enters the waste liquid pool, completing the sample quantification;

[0077] 8. The first valve is opened, and the sample in the incubation pool is mixed with the dried magnetic beads and antibodies in the antibody storage chamber. At the same time, the third valve and the fourth valve are opened, and the reagents pre-stored in the upstream reagent storage chamber, the downstream reagent storage chamber and the substrate storage chamber enter the downstream chambers. Finally, the fifth valve corresponding to the sealing liquid storage chamber is opened, and the mineral oil pre-stored in the sealing liquid storage chamber is filled into the first channel, achieving complete sealing and isolation of the downstream chamber, and then stopping centrifugation.

[0078] 9. Start heating to 37°C and keep it for 15 minutes. The magnetic beads coated with the primary antibody, the antigen in the sample and the secondary antibody will form an immune complex.

[0079] 10. The detection disk is rotated to collect the immune complexes through the magnet device provided on the detection device, and is transferred to the upstream cleaning pool through the first channel. By repeatedly rotating in forward and reverse directions, the immune complexes are completely dispersed in the upstream cleaning pool, completing the first cleaning of the immune complexes.

[0080] 11. Repeat step 10 to complete the second wash of the immune complex.

[0081] 12. Rotate the detection disk to transfer the cleaned immune complex to the reaction pool. By repeatedly rotating in forward and reverse directions, the immune complex is completely dispersed in the reaction pool. The substrate in the reaction pool reacts with the enzyme on the surface of the immune complex to generate an enzymatic reaction. The light generated is captured and recorded by the detector on the detection device.

[0082] Other preferred embodiments with different structures are similar to the above reaction process and will not be described in detail here.

[0083] The above description is only an embodiment of the present application and is not intended to limit the protection scope of the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A fully automatic magnetic microparticle chemiluminescence detection plate, characterized in that: include: Sample processing module; An incubation module, wherein the incubation module is located at a distal end side compared to the sample processing module, the incubation module comprises an incubation pool, a quantitative channel and an antibody storage cavity connected to the incubation pool via a first valve, wherein magnetic beads coated with antibodies are arranged in the antibody storage cavity, the sample processing module is connected to the proximal side of the first channel via a second valve and is located at an upper region corresponding to the incubation pool, and the distance between the proximal end of the antibody storage cavity and the rotation center is smaller than the distance between the proximal end of the incubation pool and the distal end of the quantitative channel and the rotation center; A cleaning reaction module, the cleaning reaction module is located downstream of the incubation module and is adapted to the incubation module, the cleaning reaction module comprises at least one cleaning tank and at least one reaction tank; The proximal sides of the cleaning tank, the reaction tank and the incubation tank are connected through a first channel; The first channel is in the shape of an arc or a spiral line, the incubation pool, the cleaning pool and the reaction pool are located on one side of the distal end of the first channel and are arranged in sequence along the spiral line, or the incubation pool, the cleaning pool and the reaction pool are arranged in sequence along the arc direction; The quantitative channel is connected to the waste liquid pool or the parallel incubation module; The upper region refers to the sector formed by the lines connecting the two ends of the cavity opening and the rotation center, and the area within this sector can be called the upper region.

2. The fully automatic magnetic microparticle chemiluminescence detection plate according to claim 1, characterized in that: The quantitative channel and the antibody storage cavity are respectively located on both sides of the incubation pool, the bottom of the incubation pool and the bottom of the antibody storage cavity are located on the same arc surface, and the first valve is located on the arc surface.

3. The fully automatic magnetic microparticle chemiluminescence detection plate according to claim 1, characterized in that: A reagent storage module is connected to the proximal side of the first channel through a third valve and is arranged corresponding to the upper area of ​​the cleaning tank. A substrate storage chamber is connected to the proximal side of the first channel through a fourth valve and is arranged corresponding to the upper area of ​​the reaction tank.

4. The fully automatic magnetic microparticle chemiluminescence detection plate according to claim 3, characterized in that: A sealing liquid storage chamber is connected to the proximal side of the first channel via a fifth valve.

5. The fully automatic magnetic microparticle chemiluminescence detection plate according to claim 4, characterized in that: The reagent storage module comprises a number of reagent storage chambers corresponding to the number of the cleaning tanks, the reagent storage chambers being connected to the proximal side of the first channel via a third valve and being located in the upper area of ​​the corresponding cleaning tank; or The reagent storage module includes a reagent storage chamber and a corresponding number of quantitative distribution chambers as the cleaning pool, the quantitative distribution chamber is connected to the proximal side of the first channel through a third valve and is located in the upper area of ​​the corresponding cleaning pool, the proximal end of the quantitative distribution chamber is connected in series through a distribution channel, one end of the distribution channel is connected to the proximal end of the first overflow chamber, and the other end is connected to the distal end of the reagent storage chamber through a sixth valve.

6. The fully automatic magnetic microparticle chemiluminescence detection plate according to claim 5, characterized in that: The depth of the first channel is smaller than that of the waste liquid tank, the incubation tank, the cleaning tank and the reaction tank.

7. The fully automatic magnetic microparticle chemiluminescence detection plate according to claim 1, characterized in that: The sample processing module comprises: A sample chamber, wherein the sample chamber has a sample addition port; Buffer storage module; A sample centrifugal quantitative module, wherein the distal end of the sample cavity is connected to the proximal end of the sample centrifugal quantitative module and the second overflow cavity through a seventh valve; A mixing chamber, the proximal end of which is connected to the buffer storage module through an eighth valve and to the sample centrifugation quantification module through a ninth valve; the distal end of the mixing chamber is connected to the proximal side of the first channel of the incubation module of the same number as the second valve through at least one second valve and the connection position is located in the upper area corresponding to the incubation pool.

8. The fully automatic magnetic microparticle chemiluminescence detection plate according to claim 7, characterized in that: The buffer storage module is a buffer storage chamber; or The buffer storage module includes a buffer storage cavity and a buffer quantitative pool, the proximal end of the buffer quantitative pool is connected to the buffer storage cavity through a tenth valve, the distal end of the buffer quantitative pool is connected to the mixing cavity through an eighth valve, and the proximal end of the buffer quantitative pool is provided with an overflow channel, and the overflow channel is connected to the second overflow cavity.

9. The fully automatic magnetic microparticle chemiluminescence detection plate according to claim 7, characterized in that: The sample centrifugal quantitative module comprises a serum quantitative pool, a sedimentation pool, and a check valve connecting the quantitative pool and the sedimentation pool, which are sequentially arranged away from the rotation center; One end of the ninth valve is connected to the distal end of the serum quantitative pool.

10. A detection device, characterized in that: The detection device comprises the fully automatic magnetic microparticle chemiluminescence detection disk as described in any one of claims 1-9.

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